Encyclopedia of

PHAR MACE UTICAL

TECH NOLOGY
Third Edition
VOLUME 6

DK9391_C000.indd 1 08/24/2006 3:14:02 PM

 Drugs and the Pharmaceutical Sciences
A comprehensive series of more than 160 volumes on all aspects of pharmaceutical science
Series Executive Editor: James Swarbrick

Herbal Supplements-Drug Interactions: Scientific Introduction to the Pharmaceutical Regulatory Surfactants and Polymers in Drug Delivery
and Regulatory Perspectives Process Martin Malmsten
Edited by: Y.W. Francis Lam, Shiew-Mei Huang and Edited by: Ira R. Berry ISBN: 0-8247-0804-0
Stephen D. Hall ISBN: 0-8247-5464-6
ISBN: 0-8247-2538-7 Modern Pharmaceutics, Fourth Edition
New Drug Development: Regulatory Paradigms for Edited by: Gilbert S. Banker and
Dose Optimization In Drug Development Clinical Pharmacology and Biopharmaceutics Christopher T. Rhodes
Edited by: Rajesh Krishna Edited by: Chandrah Sahajwalla ISBN: 0-8247-0674-9
ISBN: 1-5744-4808-0 ISBN: 0-8247-5465-4
Handbook of Pharmaceutical Analysis
Spectroscopy of Pharmaceutical Solids Freeze-Drying/Lyophilization of Pharmaceutical Edited by: Lena Ohannesian
Edited by: Harry G. Brittain and Biological Products, Second Edition ISBN: 0-8247-0462-2
ISBN: 1-5744-4893-5 Edited by: Louis Rey and Joan C. May
ISBN: 0-8247-4868-9 Drug-Drug Interactions
Nanoparticle Technology for Drug Delivery Edited by: A. David Rodrigues
Edited by: Ram B. Gupta and Uday B. Kompella Pharmaceutical Statistics, Fourth Edition ISBN: 0-8247-0283-2
ISBN: 1-5744-4857-9 Sanford Bolton and Charles Bon
ISBN: 0-8247-4695-3 Handbook of Drug Screening
Microencapsulation: Methods and Industrial Edited by: Ramakrishn Seethala
Applications, Second Edition Pharmaceutical Inhalation Aerosol Technology, ISBN: 0-8247-0562-9
Edited by: Simon Benita Second Edition
ISBN: 0-8247-2317-1 Edited by: Anthony J. Hickey Pharmaceutical Process Engineering
ISBN: 0-8247-4253-2 Edited by: Anthony J. Hickey
Pharmaceutical Process Scale-Up, Second Edition ISBN: 0-8247-0298-0
Edited by: Michael Levin Biomarkers in Clinical Drug Development
ISBN: 1-5744-4876-5 Edited by: John C. Bloom and Richard A. Dean Drug Stability, Third Edition, Revised, and
ISBN: 0-8247-4026-2 Expanded: Principles and Practices
Pharmacogenomics, Second Edition Edited by: Jens T. Carstensen
Edited by: Werner Kalow, Urs B. Meyer, and Rachel F. Pharmaceutical Extrusion Technology ISBN: 0-8247-0376-6
Tyndale Edited by: Isa Ghebre-Selassie and Charles Martin
ISBN: 1-5744-4878-1 ISBN: 0-8247-4050-5 Coming Soon!
November 2006
Handbook of Pharmaceutical Granulation Pharmaceutical Gene Delivery Systems Environmental Monitoring for Cleanrooms and
Technology, Second Edition Edited by: Alain Rolland and Sean M. Sullivan Controlled Environments
Edited by: Dilip M. Parikh ISBN: 0-8247-4235-4 Edited by: Anne Marie Dixon
ISBN: 0-8247-2647-2 ISBN: 0-8247-2359-7
Pharmaceutical Process Validation:
Preclinical Drug Development An International Third Edition Pharmaceutical Photostability and Stabilization
Edited by: Mark C. Rogge Edited by: Robert A. Nash and Alfred H. Wachter Technology
ISBN: 1-5744-4882-X ISBN: 0-8247-0838-5 Edited by: Joseph T. Piechocki
ISBN: 0-8247-5924-9
Percutaneous Absorption: Drugs, Cosmetics, Ophthalmic Drug Delivery Systems, Second
Mechanisms, Methods, Fourth Edition Edition December 2006
Edited by: Robert L. Bronaugh and Howard I. Maibach Edited by: Ashim K. Mitra Good Manufacturing Practices for
ISBN: 1-5744-4869-2 ISBN: 0-8247-4124-2 Pharmaceuticals: A Plan for Total Quality
Control from Manufacturer to Consumer, Sixth
Pharmaceutical Stress Testing: Predicting Drug Affinity Capillary Electrophoresis in Pharmaceutics Edited by: Joseph Nally
Degradation and Biopharmaceutics ISBN: 0-8493-3972-3
Edited by: Steven W. Baertschi Edited by: Reinhard Neubert
ISBN: 0-8247-4021-1 ISBN: 0-8247-0951-9 February 2007
Pharmaceutical Product Development: In Vitro-In
Active Pharmaceutical Ingredients: Development, Parenteral Quality Control: Sterility, Pyrogen, Vivo Correlation
Manufacturing, and Regulation Particulate, and Package Integrity Testing: Edited by: Dakshina Chilukuri
Edited by: Stanley Nusim Third Edition ISBN: 0-8493-3827-1
ISBN: 0-8247-0293-X Michael K. Akers, Daniel S. Larrimore, and Dana
Morton Guazzo Endotoxins: Pyrogens, LAL Testing and
Injectable Dispersed Systems: Formulation, ISBN: 0-8247-0885-7 Depyrogenation, Third Edition
Processing and Performance Kevin L. Williams
Edited by: Diane J. Burgess Modified-Release Drug Delivery Technology ISBN: 0-8493-9372-8
ISBN: 0-8493-3699-6 Edited by: Michael J. Rathbone
ISBN: 0-8247-0869-5 April 2007
Polymeric Drug Delivery Systems Good Laboratory Practice Regulations,
Edited by: Glen S. Kwon Simulation for Designing Clinical Trials: A Fourth Edition
ISBN: 0-8247-2532-8 Pharmacokinetic-Pharmacodynamic Modeling Edited by: Sandy Weinberg
Perspective ISBN: 0-8493-7583-5
Drug Delivery to the Oral Cavity Edited by: Hui Kimko
Edited by: Tapash K. Ghosh and William Pfister ISBN: 0-8247-0862-8 June 2007
ISBN: 0-8247-8293-3 Filtration and Purification in the Biopharmaceutical
Transdermal Drug Delivery Industry, Second Edition
Generic Drug Development: Solid Oral Dosage Forms Second Edition Edited by: Theodore H. Meltzer and Maik Jornitz
Edited by: Leon Shargel Edited by: Jonathan Hadgraft ISBN: 0-8493-7953-9
ISBN: 0-8247-5460-3 ISBN: 0-8247-0861-X

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 Encyclopedia of
PHAR MACE UTICAL
TECH NOLOGY
Third Edition
VOLUME 6

edited by
James Swarbrick
PharmaceuTech, Inc.
Pinehurst, North Carolinia, USA

New York London

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 Informa Healthcare USA, Inc.
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 To James C. Boylan
Co-Editor of the First and Second Editions
A great colleague and friend.
 Editorial Advisory Board
Jean-Marc Aiache Igor Gonda
Biopharmaceutics Department, University of Clermont Acrux Limited, West Melbourne,
Ferrand, Clermont Ferrand, France Victoria, Australia

David Attwood Anthony J. Hickey

School of Pharmacy & Pharmaceutical Sciences, Department of Pharmaceutics, School of Pharmacy,
University of Manchester, Manchester, U.K. University of North Carolina at Chapel Hill,
Chapel Hill, North Carolina, U.S.A.
Larry L. Augsburger
Department of Pharmaceutics, University of Maryland Hideki Ichikawa
at Baltimore, Baltimore, Maryland, U.S.A. Faculty of Pharmaceutical Sciences,
Kobe Gakuin University, Kobe, Japan
Roland A. Bodmeier
College of Pharmacy, Freie Universität Berlin, Brian E. Jones
Berlin, Germany Qualicaps, S.A., Penarth, U.K.

J. Phillip Bowen James C. McElnay

Center for Biomolecular Structure and Dynamics, Faculty of Science and Agriculture, Queen’s University
Department of Chemistry, University of Georgia, Belfast, Belfast, U.K.
Athens, Georgia, U.S.A.
Iain J. McGilveray
Gayle A. Brazeau McGilveray Pharmacon Inc., Nepean,
School of Pharmacy, State University of New York at Ontario, Canada
Buffalo, Amherst, New York, U.S.A.
Kinam Park
Harry G. Brittain School of Pharmacy and Pharmacal Sciences, Purdue
Center for Pharmaceutical Physics, Milford, University, West Lafayette, Indiana, U.S.A.
New Jersey, U.S.A.
Michael J. Pikal
Les F. Chasseaud School of Pharmacy, University of Connecticut, Storrs,
Drug Metabolism and Pharmacokinetics (DMPK), Connecticut, U.S.A.
Cambridge, U.K.
Naı́r Rodrı́guez-Hornedo
Yie W. Chien Department of Pharmaceutical Sciences, College of
Research and Development, Kaohsiung Medical Pharmacy, University of Michigan, Ann Arbor,
University, Kaohsiung, Taiwan Michigan, U.S.A.

Bradley A. Clark Stephen G. Schulman

Solvay Pharmaceuticals, Marietta, Georgia, U.S.A. Department of Medicinal Chemistry, College of
Pharmacy, University of Florida, Gainesville,
Owen I. Corrigan Florida, U.S.A.
Department of Pharmaceutics, School of Pharmacy,
University of Dublin, Trinity College, Dublin, Ireland Jerome P. Skelly
Pharmaceutical Consultant, Alexandria,
Gillian M. Eccleston Virginia, U.S.A.
Department of Pharmaceutical Sciences, University of
Strathclyde, Glasgow, Scotland, U.K. Clive G. Wilson
Department of Pharmaceutical Sciences,
James L. Ford Strathclyde Institute for Biomedical Sciences (SIBS),
School of Pharmacy and Chemistry, Liverpool John University of Strathclyde,
Moores University, Liverpool, U.K. Glasgow, U.K.

vii
Contributors

Ragheb Abu-Rmaileh = School of Pharmacy and Pharmaceutical Sciences, University of Manchester,

Manchester, U.K.
Phillip M. Achey = Department of Microbiology and Cell Science, Institute of Food and
Agricultural Sciences, University of Florida, Gainesville, Florida, U.S.A.
James P. Agalloco = Agalloco and Associates Inc., Belle Mead, New Jersey, U.S.A.
Vikas Agarwal = CIMA Labs, Inc., Brooklyn Park, Minnesota, U.S.A.
Alaa M. Ahmad = Department of Clinical Pharmacology, Vertex Pharmaceuticals, Inc.,
Cambridge, Massachusetts, U.S.A.
Jean-Marc Aiache = Biopharmaceutics Department, University of Clermont-Ferrand,
Clermont Ferrand, France
James E. Akers = Akers Kennedy and Associates, Kansas City, Missouri, U.S.A.
Michael J. Akers = Martinsville, Indiana, U.S.A.
Hemant H. Alur = Division of Pharmaceutical Sciences, School of Pharmacy,
University of Missouri-Kansas City, Kansas City, Missouri, U.S.A.
David G. Allison = School of Pharmacy and Pharmaceutical Sciences, University of Manchester,
Manchester, U.K.
Norman Anthony Armstrong = Welsh School of Pharmacy, Cardiff University, Cardiff, U.K.
Agnès Artiges = European Directorate for the Quality of Medicines (EDQM),
Strasbourg Cedex, France
Carolyn H. Asbury = Department of Medicine, University of Pennsylvania Health System,
Philadelphia, Pennsylvania, U.S.A.
C. K. Atterwill = Biosciences Division, Huntingdon Life Sciences, Cambridgeshire, U.K.
David Attwood = School of Pharmacy and Pharmaceutical Sciences, University of Manchester,
Manchester, U.K.
Larry L. Augsburger = Department of Pharmaceutics, University of Maryland at Baltimore,
Baltimore, Maryland, U.S.A.
J. Desmond Baggot = School of Veterinary Medicine, St. George’s University, Grenada, West Indies
Mark L. Balboni = KMI, A Division of PAREXEL International, LLC, Waltham,
Massachusetts, U.S.A.
Paul Baldrick = Covance Laboratories, Ltd., North Yorkshire, U.K.
Samuel B. Balik = GlaxoSmithKline, Research Triangle Park, North Carolina, U.S.A.
David J. Barlow = Molecular Biophysics Group, Pharmaceutical Science Research Division,
King’s College London, College London, London, U.K.
Debra Barnes = Roche Global Development, Palo Alto, California, U.S.A.
Onkaram Basavapathruni = Pharmacia, Skokie, Illinois, U.S.A.
Annette Bauer-Brandl = Institute of Pharmacy, Department of Pharmaceutics and Biopharmaceutics,
The University of Tromsø, Tromsø, Norway
G.J.P.J. Beernink = Amsterdam, The Netherlands
Leslie Z. Benet = Department of Biopharmaceutical Sciences, University of California
San Francisco, San Francisco, California, U.S.A.
David H. Bergstrom = Cardinal Health Pharmaceutical Development, Somerset, New Jersey, U.S.A.
Ira R. Berry = International Regulatory Business Consultants, LLC, Maplewood, New Jersey, U.S.A.

ix
 x

Guru Betageri = College of Pharmacy, Western University of Health Sciences,

Pomona, California, U.S.A.
Erick Beyssac = Biopharmaceutical Department, Faculty of Pharmacy, University of Auvergne,
Clermont-Ferrand, France
Haresh Bhagat = Alcon Research, Ltd., Fort Worth, Texas, U.S.A.
Hridaya Bhargava = Massachusetts College of Pharmacy and Health Sciences,
Boston, Massachusetts, U.S.A.
Marı́a Dolores Blanco = Departamento de Bioquimica y Biologia Molecular,
Universidad Complutense de Madrid, Madrid, Spain
Maria Jose Blanco-Prı́eto = University of Navarra, Pamplona, Spain
E. Daniel Blankschtein = Department of Chemical Engineering, Massachusetts Institute of Technology,
Cambridge, Massachusetts, U.S.A.
Roland A. Bodmeier = College of Pharmacy, Freie Universität Berlin, Berlin, Germany
Michael J. Bogda = Barr Laboratories, Inc., Woodcliff Lake, New Jersey, U.S.A.
René Bommer = Ing. Erich Pfeiffer GmbH, Radolfzell, Germany
Charles Bon = AAI Pharma, Inc., Wilmington, North Carolina, U.S.A.
Carol A. Borynec = Capital Health, Edmonton, Alberta, Canada
David W.A. Bourne = College of Pharmacy, Health Sciences Center, The University of Oklahoma,
Oklahoma City, Oklahoma, U.S.A.
J. Phillip Bowen = Center for Biomolecular Structure and Dynamics, University of Georgia,
Athens, Georgia, U.S.A.
Richard D. Braatz = Department of Chemical and Biomolecular Engineering, University of Illinois at
Urbana-Champaign, Urbana, Illinois, U.S.A.
Daniel A. Brazeau = Department of Pharmaceutical Sciences, The State University of New York
at Buffalo, Buffalo, New York, U.S.A.
Gayle A. Brazeau = School of Pharmacy, The State University of New York at Buffalo,
Buffalo, New York, U.S.A.
Ron J. Brendel = Mallinckrodt, Inc., St. Louis, Missouri, U.S.A.
Harry G. Brittain = Center for Pharmaceutical Physics, Milford, New Jersey, U.S.A.
Albert W. Brzeczko = APC/Niro Inc., Columbia, Maryland, U.S.A.
Robert A. Buerki = College of Pharmacy, The Ohio State University,
Columbus, Ohio, U.S.A.
Diane J. Burgess = Department of Pharmaceutical Sciences, University of Connecticut,
Storrs, Connecticut, U.S.A.
Till Bussemer = Aventis Pharma Deutschland GmbH, Frankfurt am Main, Germany
John B. Cannon = Abbott Laboratories, North Chicago, Illinois, U.S.A.
J.-M. Cardot = Biopharmaceutical Department, Faculty of Pharmacy, University of Auvergne,
Clermont-Ferrand, France
Ralph B. Caricofe = GlaxoSmithKline, Research Triangle Park, North Carolina, U.S.A.
Eugene Carstea = Invitrogen Corporation, Carlsbad, California, U.S.A.
Allen Cato = Cato Research Ltd., Durham, North Carolina, U.S.A.
Allen Cato III = Cato Research Ltd., San Diego, California, U.S.A.
Hak-Kim Chan = University of Sydney, Sydney, New South Wales, Australia
Les F. Chasseaud = Drug Metabolism and Pharmacokinetics (DMPK), Cambridge, U.K.
Xiu Xiu Cheng = Andrx Pharmaceuticals, Inc., Fort Lauderdale, Florida, U.S.A.
David L. Chesney = KMI, A Division of PAREXEL International, LLC,
Waltham, Massachusetts, U.S.A.
Nora Y.K. Chew = Acrux DDS Pty Ltd., West Melbourne, Victoria, Australia
Yie W. Chien = Research and Development, Kaohsiung Medical University,
Kaohsiung, Taiwan
Andrea Chieppo = Analytical Research Laboratories, Oklahoma City, Oklahoma, U.S.A.
 xi

Sally Y. Choe = Pfizer Global Research and Development, Ann Arbor, Michigan, U.S.A.
Masood Chowhan = Alcon Laboratories, Fort Worth, Texas, U.S.A.
Sebastian G. Ciancio = Department of Periodontics and Endodontics, The New York State University
at Buffalo, Buffalo, New York, U.S.A.
Emil W. Ciurczak = Integrated Technical Solutions, Goldens Bridge, New York, U.S.A.
Bradley A. Clark = Solvay Pharmaceuticals, Inc., Marietta, Georgia, U.S.A.
C. Randall Clark = School of Pharmacy, Auburn University, Auburn, Alabama, U.S.A.
Nigel Clarke = Schering-Plough Research Institute, Kenilworth, New Jersey, U.S.A.
Sophie-Dorothée Clas = Merck Frosst Canada and Company, Pointe Claire-Dorval,
Quebec, Canada
Sarah M.E. Cockbill = University of Wales Cardiff, Cardiff, U.K.
Douglas L. Cocks = Eli Lilly and Company, Indianapolis, Indiana, U.S.A.
Elizabeth Colbourn = Intelligensys Ltd., Billingham, Teesside, U.K.
James J. Conners = Sepracor, Marlborough, Massachusetts, U.S.A.
Kenneth A. Connors = Center for Health Sciences, University of Wisconsin at Madison,
Madison, Wisconsin, U.S.A.
Antonio Conto = ChemSafe S.a.s., Colleretto Giacosa (TO), Italy
Chyung S. Cook = Pharmacia Corporation, Skokie, Illinois, U.S.A.
James F. Cooper = Charles River Endosafe, Charleston, South Carolina, U.S.A.
Geoffrey A. Cordell = College of Pharmacy, University of Illinois at Chicago,
Chicago, Illinois, U.S.A.
Owen I. Corrigan = Trinity College, University of Dublin, Dublin, Ireland
Michael Cory = GlaxoSmithKline, Research Triangle Park, North Carolina, U.S.A.
Diane D. Cousins = U.S. Pharmacopeial Convention, Inc., Rockville, Maryland, U.S.A.
Kathleen A. Cox = Schering-Plough Research Institute, Kenilworth, New Jersey, U.S.A.
Charles W. Crew = GlaxoSmithKline, Research Triangle Park, North Carolina, U.S.A.
Alan L. Cripps = GlaxoSmithKline, Hertfordshire, U.K.
Mary E.M. Cromwell = Genentech, Inc., South San Francisco, California, U.S.A.
Patrick J. Crowley = GlaxoSmithKline, Essex, U.K.
Anthony M. Cundell = Microbiological Development and Statistics, Wyeth-Ayerst Pharmaceuticals,
Pearl River, New York, U.S.A.
Chad R. Dalton = Merck Frosst Canada and Company, Pointe Claire-Dorval,
Quebec, Canada
James T. Dalton = Division of Pharmaceutics, College of Pharmacy, The Ohio State University,
Columbus, Ohio, U.S.A.
Wenbin Dang = Cardinal Health Pharmaceutical Development, Somerset, New Jersey, U.S.A.
Ira Das = St. Louis, Missouri, U.S.A.
Judith A. Davis = Department of Pharmacy Health Care Administration, College of Pharmacy,
University of Florida, Gainesville, Florida, U.S.A.
Michael R. DeFelippis = Lilly Research Laboratories, Eli Lilly and Company, Indianapolis,
Indiana, U.S.A.
M. Begon~a Delgado-Charro = Centre Interuniversitaire de Recherche et d’Enseignement,
Universities of Geneva and Lyon, Archamps, Switzerland
Antony D’Emanuele = School of Pharmacy and Pharmaceutical Sciences, University of Manchester,
Manchester, U.K.
Nigel J. Dent = Country Consultancy Ltd., Milton Malsor, U.K.
Jack DeRuiter = School of Pharmacy, Auburn University, Auburn, Alabama, U.S.A.
Jeffrey Ding = Battelle Pulmonary Therapeutics, Inc., Columbus, Ohio, U.S.A.
Marilyn D. Duerst = Department of Chemistry, University of Wisconsin-River Falls,
River Falls, Wisconsin, U.S.A.
 xii

A. Mark Dyas = School of Pharmacy and Chemistry, Liverpool John Moores University,
Liverpool, U.K.
Stefan Eberl = Department of PET and Nuclear Medicine, Royal Prince Alfred Hospital,
Sydney, New South Wales, Australia
Gillian M. Eccleston = Department of Pharmaceutical Sciences, Strathclyde Institute
for Biomedical Sciences, Glasgow, Scotland, U.K.
Joachim Ermer = Aventis Pharma AG, Frankfurt am Main, Germany
Ene I. Ette = Department of Clinical Pharmacology, Vertex Pharmaceuticals, Inc.,
Cambridge, Massachusetts, U.S.A.
Ronald P. Evens = MAPS 4 Biotec, Inc., Jacksonville, Florida, U.S.A.
Kevin L. Facchine = GlaxoSmithKline, Research Triangle Park, North Carolina, U.S.A.
Gordon J. Farquharson = Bovis Tanvec, Ltd., Tanshire House, Surrey, U.K.
Elias Fattal = School of Pharmacy, University of Paris XI, Cha ^tenay-Malabry, France
Linda A. Felton = College of Pharmacy, University of New Mexico, Albuquerque, New Mexico, U.S.A.
Charles W. Fetrow = St. Francis Medical Center, Pittsburgh, Pennsylvania, U.S.A.
John W.A. Findlay = Pfizer Global Research and Development, Groton, Connecticut, U.S.A.
Barrie C. Finnin = Monash University, Parkville, Victoria, Australia
Elizabeth S. Fisher = Merck and Co., Inc., Rahway, New Jersey, U.S.A.
Joseph A. Fix = Yamanouchi Pharma Technologies, Inc., Palo Alto, California, U.S.A.
Farrel L. Fort = TAP Pharmaceutical Products, Inc., Lake Forest, Illinois, U.S.A.
Miriam K. Franchini = Bristol-Myers Squibb, New Brunswick, New Jersey, U.S.A.
Cara R. Frosch = Covance Central Laboratory Services Inc., Indianapolis, Indiana, U.S.A.
Mitsuko Fujiwara = Department of Chemical and Biomolecular Engineering, University of Illinois
at Urbana-Champaign, Urbana, Illinois, U.S.A.
Yoshinobu Fukumori = Kobe Gakuin University, Kobe, Hyogo, Japan
Kumar G. Gadamasetti = ChemRx Advanced Technologies, South San Francisco, California, U.S.A.
Bruno Gander = Institute of Pharmaceutical Sciences, ETH Zürich, Zürich, Switzerland
David Ganderton = Devon, U.K.
Isaac Ghebre-Sellassie = Pharmaceutical Technology Solutions, Morris Plains, New Jersey, U.S.A.
Peter J. Giddings = SmithKline Beecham, Brentford, U.K.
Peter Gilbert = University of Manchester, Manchester, U.K.
Danièlle Giron = Analytical Research and Development, Sandoz Pharma, Basel, Switzerland
Samuel V. Givens = Hoffmann-La Roche, Inc., Nutley, New Jersey, U.S.A.
William Glover = University of Sydney, Sydney, New South Wales, Australia
Igor Gonda = Acrux Limited, West Melbourne, Australia
Michelle M. Gonzalez = Amgen Inc., Thousand Oaks, California, U.S.A.
Lee T. Grady = McLean, Virginia, U.S.A.
Alice T. Granger = Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, Connecticut, U.S.A.
Jerome J. Groen = Abbott Laboratories, Abbott Park, Illinois, U.S.A.
Richard A. Guarino = Oxford Pharmaceutical Resources, Inc., Totowa, New Jersey, U.S.A.
Pramod K. Gupta = Baxter Healthcare Corporation, Round Lake, Illinois, U.S.A.
Richard H. Guy = Centre Interuniversitaire de Recherche et d’Enseignement, Universities of Geneva
and Lyon, Archamps, Switzerland
J. Richard Gyory = ALZA Corporation, Spring Lake Park, Minnesota, U.S.A.
Huijeong Ashley Hahm = Office of Generic Drugs, U.S. Food and Drug Administration, Rockville,
Maryland, U.S.A.
Nigel A. Halls = NHC-Nigel Halls Consulting, Herts, U.K.
Jerome A. Halperin = Silver Spring, Maryland, U.S.A.
Bruno C. Hancock = Pfizer Inc., Groton, Connecticut, U.S.A.
 xiii

Henri Hansson = Galenica AB, Medeon, Malmo, Sweden

Gary C. Harbour = Pharmacia Enterprises S.A., Luxembourg, G. D. Luxembourg
Adam Harris = Invitrogen Corporation, Carlsbad, California, U.S.A.
D. R. Hawkins = Huntingdon Life Sciences, Cambridgeshire, U.K.
Leslie C. Hawley = Pharmacia Corporation, Kalamazoo, Michigan, U.S.A.
Anne Marie Healy = Department of Pharmaceutics, School of Pharmacy, University of Dublin,
Dublin, Ireland
Herbert Michael Heise = University of Dortmund, Dortmund, Germany
Paul W.S. Heng = Department of Pharmacy, National University of Singapore, Singapore
Jeffrey M. Herz = Omeros Medical Systems, Inc., Seattle, Washington, U.S.A.
Anthony J. Hickey = Department of Pharmaceutics, The University of North Carolina at Chapel Hill,
Chapel Hill, North Carolina, U.S.A.
Gregory J. Higby = American Institute of the History of Pharmacy, Madison, Wisconsin, U.S.A.
Anthony J. Hlinak = Searle Research and Development, Skokie, Illinois, U.S.A.
Amnon Hoffman = Department of Pharmaceutics, Pharmacy School, The Hebrew University of
Jerusalem, Jerusalem, Israel
Harm HogenEsch = Department of Veterinary Pathobiology, Purdue University,
West Lafayette, Indiana, U.S.A.
R. Gary Hollenbeck = Department of Pharmaceutics, University of Maryland at Baltimore,
Baltimore, Maryland, U.S.A.
J. Hotta = Bayer Corporation, Research Triangle Park, North Carolina, U.S.A.
Shelley Hough = Invitrogen Corporation, Carlsbad, California, U.S.A.
Stephen A. Howard = Purdue Pharma L.P., Ardsley, New York, U.S.A.
Carmel M. Hughes = School of Pharmacy, Medical Biology Centre, The Queen’s University of Belfast,
Belfast, U.K.
Jeffrey A. Hughes = College of Pharmacy, University of Florida, Gainesville, Florida, U.S.A.
Kang Moo Huh = Departments of Pharmaceutics and Biomedical Engineering, Purdue University,
West Lafayette, Indiana, U.S.A.
Ho-Wah Hui = Abbott Laboratories, North Chicago, Illinois, U.S.A.
Laura Hungerford = U.S. Food and Drug Administration, Rockville, Maryland, U.S.A.
D. Hunkeler = Laboratoire des Polye´lectrolytes et BioMacromole´cules,
EPFL, Lausanne, Switzerland
Desmond G. Hunt = Department of Standards Development, United States Pharmacopeia,
Rockville, Maryland, U.S.A.
Anwar A. Hussain = Department of Pharmaceutics, University of Kentucky,
Lexington, Kentucky, U.S.A.
Daniel A. Hussar = University of the Sciences in Philadelphia, Philadelphia, Pennsylvania, U.S.A.
Hideki Ichikawa = Kobe Gakuin University, Kobe, Hyogo, Japan
Juhana E. Idanpaan-Heikkila = World Health Organization, Geneva, Switzerland
Victoria Imber = Evergreen, Colorado, U.S.A.
Barbara Immel = Immel Resources, Petaluma, California, U.S.A.
David M. Jacobs = European Agency for the Evaluation of Medicinal Products, Basel, Switzerland
Adivaraha Jayasankar = Department of Pharmaceutical Sciences, University of Michigan,
Ann Arbor, Michigan, U.S.A.
Bradford K. Jensen = Aventis Pharmaceuticals, Inc., Bridgewater, New Jersey, U.S.A.
Thomas P. Johnston = Division of Pharmaceutical Sciences, School of Pharmacy, University of
Missouri-Kansas City, Kansas City, Missouri, U.S.A.
Brian E. Jones = Qualicaps Europe, S.A., Alcobendas, Madrid, Spain
Deborah J. Jones = Steripak Limited, Cheshire, U.K.
Maik W. Jornitz = Sartorius Group, Goettingen, Germany
 xiv

Jose C. Joseph = Abbott Laboratories, Abbott Park, Illinois, U.S.A.

Hans E. Junginger = Division of Pharmaceutical Technology, Leiden/Amsterdam Center for Drug
Research, Leiden, The Netherlands
Galina N. Kalinkova = Department of Pharmaceutical Chemistry, Medical University, Sofia, Bulgaria
Werner Kalow = Department of Pharmacology, University of Toronto,
Toronto, Ontario, Canada
Fumio Kamiyama = Department of Biopharmaceutics, Kyoto Pharmaceutical University,
Kyoto, Japan
Isadore Kanfer = Rhodes University, Grahamstown, South Africa
Aziz Karim = Pharmacia Corporation, Skokie, Illinois, U.S.A.
Brian H. Kaye = Department of Physics and Astronomy, Laurentian University,
Sudbury, Ontario, Canada
Ron C. Kelly = SSCI, Inc., West Lafayette, Indiana, U.S.A.
György M. Keserü = Computer Assisted Drug Discovery and HTS Unit, Gedeon Richter Ltd.,
Budapest, Pest, Hungary
David P. Kessler = Department of Chemical Engineering, Purdue University,
West Lafayette, Indiana, U.S.A.
Rajendra K. Khankari = CIMA Labs, Inc., Brooklyn Park, Minnesota, U.S.A.
Ban-An Khaw = Center for Cardiovascular Targeting, School of Pharmacy, Northeastern University,
Boston, Massachusetts, U.S.A.
Arthur H. Kibbe = Department of Pharmaceutical Sciences, Wilkes University, Wilkes-Barre,
Pennsylvania, U.S.A.
Robert G. Kieffer = RGK Consulting, Palm Desert, California, U.S.A.
Chris C. Kiesnowski = Bristol-Myers Squibb Pharmaceutical Research Institute, New Brunswick,
New Jersey, U.S.A.
Kwon H. Kim = Department of Pharmacy and Administrative Sciences, College of Pharmacy and
Allied Health Professions, St. John’s University, Jamaica, New York, U.S.A.
Toby King = Aradigm Corporation, Hayward, California, U.S.A.
Florence K. Kinoshita = Hercules Incorporated, Wilmington, Delaware, U.S.A.
Cathy M. Klech-Gelotte = McNeil Consumer Healthcare, Fort Washington, Pennsylvania, U.S.A.
A. Klos = Bayer Corporation, Research Triangle Park, North Carolina, U.S.A.
Axel Knoch = Pfizer Global Research and Development, Freiburg, Germany
John J. Koleng, Jr. = College of Pharmacy, University of Texas at Austin, Austin, Texas, U.S.A.
Uday B. Kompella = Department of Pharmaceutical Sciences, University of Nebraska Medical Center,
Omaha, Nebraska, U.S.A.
Sheldon X. Kong = Merck and Co., Inc., Whitehouse Station, New Jersey, U.S.A.
Mark J. Kontny = Pharmacia, Skokie, Illinois, U.S.A.
Bhavesh H. Kothari = CIMA Labs, Inc., Brooklyn Park, Minnesota, U.S.A.
Rajesh Krishna = Aventis Pharmaceuticals, Inc., Bridgewater, New Jersey, U.S.A.
Amit Kulkarni = Department of Clinical Sciences and Administration, College of Pharmacy,
University of Houston, Houston, Texas, U.S.A.
Thomas Kupiec = Analytical Research Laboratories, Oklahoma City, Oklahoma, U.S.A.
Philip Chi Lip Kwok = University of Sydney, Sydney, New South Wales, Australia
Sylvie Laganière = Integrated Research, Inc., Montreal, Quebec, Canada
Duane B. Lakings = DSE Consulting, Birmingham, Alabama, U.S.A.
John S. Landy = Aventis Pharmaceuticals, Bridgewater, New Jersey, U.S.A.
Robert Langer = Department of Chemical Engineering, Massachusetts Institute of Technology,
Cambridge, Massachusetts, U.S.A.
M. Jayne Lawrence = Molecular Biophysics Group, Pharmaceutical Science Research Division,
King’s College London, College London, London, U.K.
Destin A. LeBlanc = Cleaning Validation Technologies, San Antonio, Texas, U.S.A.
 xv

Chao-Pin Lee = GlaxoSmithKline Pharmaceuticals, Collegeville, Pennsylvania, U.S.A.

Chi H. Lee = School of Pharmacy, Division of Pharmaceutical Sciences, University of Missouri,
Kansas City, Kansas City, Missouri, U.S.A.
Kyung Hee Lee = College of Pharmacy, University of Illinois at Chicago,
Chicago, Illinois, U.S.A.
Mike S. Lee = Milestone Development Services, Newtown, Pennsylvania, U.S.A.
Sang Cheon Lee = Korea Institute of Ceramic Engineering and Technology, Seoul, South Korea
Vincent H.L. Lee = Department of Pharmaceutical Sciences, University of Southern California,
Los Angeles, California, U.S.A.
Kristi L. Lenz = Department of Pharmacy Practice, Medical University of South Carolina,
Charleston, South Carolina, U.S.A.
Jason M. LePree = Roche Pharmaceuticals, Inc., Nutley, New Jersey, U.S.A.
Michael Levin = Metropolitan Computing Corporation, East Hanover, New Jersey, U.S.A.
Gareth A. Lewis = Sanofi-Synthelabo Research, Chilly Mazarin, France
Luk Chiu Li = HPD Advanced Drug Delivery, Abbott Park, Illinois, U.S.A.
S. Kevin Li = Department of Pharmaceutics and Pharmaceutical Chemistry, College of Pharmacy,
University of Utah, Salt Lake City, Utah, U.S.A.
Yu Li = GlaxoSmithKline Pharmaceuticals, Collegeville, Pennsylvania, U.S.A.
Eric J. Lien = Biomedicinal Chemistry and Pharmaceutics, University of Southern California,
Los Angeles, California, U.S.A.
Senshang Lin = College of Pharmacy and Allied Health Professions, St. John’s University,
Jamaica, New York, U.S.A.
Nils-Olof Lindberg = Pharmacia and Upjohn AB, Helsingborg, Sweden
Blythe Lindsay = Department of Pharmaceutical Sciences, University of Strathclyde,
Glasgow, U.K.
Thomas Linz = Schering AG, Berlin, Germany
John M. Lipari = Abbott Laboratories, North Chicago, Illinois, U.S.A.
Linda Lisgarten = Library and Information Services, School of Pharmacy, University of London,
London, U.K.
G. Brian Lockwood = University of Manchester, Manchester, U.K.
Orlando Lopez = Johnson & Johnson, Raritan, New Jersey, U.S.A.
Joseph W. Lubach = Department of Pharmaceutical Chemistry, University of Kansas,
Lawrence, Kansas, U.S.A.
Craig E. Lunte = Department of Chemistry, University of Kansas, Lawrence, Kansas, U.S.A.
Robert L. Maher, Jr. = Duquesne University, Pittsburgh, Pennsylvania, U.S.A.
Joseph Maida = Maida Engineering, Inc., Fort Washington, Pennsylvania, U.S.A.
Henri R. Manasse, Jr. = American Society of Health-System Pharmacists, Bethesda,
Maryland, U.S.A.
Laviero Mancinelli = Daly City, California, U.S.A.
Peter Markland = Southern Research Institute, Birmingham, Alabama, U.S.A.
Diego Marro = Centre Interuniversitaire de Recherche et d’Enseignement,
Universities of Geneva and Lyon, Archamps, Switzerland
Marilyn N. Martinez = U.S. Food and Drug Administration, Rockville, Maryland, U.S.A.
Luigi G. Martini = GlaxoSmithKline, Essex, U.K.
Michael B. Maurin = QS Pharma, Boothwyn, Pennsylvania, U.S.A.
Joachim Mayer = Institute of Medicinal Chemistry, University of Lausanne,
Lausanne, Switzerland
Aaron S. Mayo = Department of Pharmaceutical Sciences, University of Nebraska Medical Center,
Omaha, Nebraska, U.S.A.
Orla McCallion = Vandsons Research, London, U.K.
James C. McElnay = Queen’s University Belfast, Belfast, U.K.
 xvi

Iain J. McGilveray = McGilveray Pharmacon, Inc., Nepean, Ontario, Canada

Jim W. McGinity = College of Pharmacy, University of Texas at Austin, Austin, Texas, U.S.A.
Michael McKenna = Pfizer Pharmaceuticals Group, New York, New York, U.S.A.
Marghi R. McKeon = Lab Safety Corporation, Des Plaines, Illinois, U.S.A.
Eugene J. McNally = Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, Connecticut, U.S.A.
Duncan E. McVean = Ben Venue Laboratories, Solon, Ohio, U.S.A.
Mark R. Mecadon = Novartis Pharmaceuticals, East Hanover, New Jersey, U.S.A.
Thomas Medwick = Department of Pharmaceutical Chemistry, Rutgers University, New Brunswick,
New Jersey, U.S.A.
Robert W. Mendes = Dedham, Massachusetts, U.S.A.
Jonathan M. Miller = Department of Pharmaceutical Sciences, University of Michigan,
Ann Arbor, Michigan, U.S.A.
Ronald W. Miller = Bristol-Myers Squibb, New Brunswick, New Jersey, U.S.A.
Ashim K. Mitra = Division of Pharmaceutical Sciences, School of Pharmacy,
University of Missouri-Kansas City, Kansas City, Missouri, U.S.A.
Samir S. Mitragotri = Department of Chemical Engineering, University of California,
Santa Barbara, Santa Barbara, California, U.S.A.
Suresh K. Mittal = Department of Veterinary Pathobiology, Purdue University,
West Lafayette, Indiana, U.S.A.
Derek V. Moe = CIMA Labs, Inc., Brooklyn Park, Minnesota, U.S.A.
Nicholas Mohr = Department of Emergency Medicine, Indiana University School of Medicine,
Indianapolis, Indiana, U.S.A.
Matthew J. Mollan, Jr. = Pfizer Global Research and Development, Ann Arbor,
Michigan, U.S.A.
Lorie Ann Morgan = GlaxoSmithKline, Research Triangle Park, North Carolina, U.S.A.
Karen Morisseau = College of Pharmacy, University of Rhode Island,
Kingston, Rhode Island, U.S.A.
Takahiro Morita = Tanabe Pharmaceuticals Co., Ltd., Osaka, Japan
Gerold Mosher = CyDex, Inc., Overland Park, Kansas, U.S.A.
Ronald L. Mueller = GlaxoSmithKline, King of Prussia, Pennsylvania, U.S.A.
Christel C. Mueller-Goymann = Institut für Pharmazeutische Technologie, Technischen Universität
Braunschweig, Braunschweig, Germany
Suman K. Mukherjee = Department of Pharmaceutical Sciences, South Dakota State University,
Brookings, South Dakota, U.S.A.
James O. Mullis = Department of Analytical Sciences, Boehringer Ingelheim Pharmaceuticals, Inc.,
Ridgefield, Connecticut, U.S.A.
Sandy J.M. Munro = GlaxoSmithKline, Hertfordshire, U.K.
Eric J. Munson = Department of Pharmaceutical Chemistry, University of Kansas, Lawrence,
Kansas, U.S.A.
Thomas D. Murphy = Morristown, New Jersey, U.S.A.
Fernando J. Muzzio = Department of Chemical and Biochemical Engineering, Rutgers University,
Piscataway, New Jersey, U.S.A.
Paul B. Myrdal = Department of Pharmacy Practice and Science, College of Pharmacy,
The University of Arizona, Tucson, Arizona, U.S.A.
Venkatesh Naini = Barr Laboratories, Inc., Pomona, New York, U.S.A.
Jintana Napaporn = Department of Pharmaceutics, College of Pharmacy, University of Florida,
Gainesville, Florida, U.S.A.
Robert A. Nash = Consultant, Mahwah, New Jersey, U.S.A.
Sarah J. Nehm = Department of Pharmaceutical Sciences, University of Michigan,
Ann Arbor, Michigan, U.S.A.
Deanna J. Nelson = BioLink Life Sciences, Inc., Cary, North Carolina, U.S.A.
 xvii

Sandeep Nema = Pharmacia Corp., Skokie, Illinois, U.S.A.

Michael T. Newhouse = Inhale Therapeutic Systems, Inc., San Carlos, California, U.S.A.
Ann W. Newman = SSCI, Inc., West Lafayette, Indiana, U.S.A.
J. M. Newton = Department of Pharmaceutics, University of London, London, U.K.
Daniel L. Norwood = Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, Connecticut, U.S.A.
Thomas M. Nowak = Abbott Laboratories, Abbott Park, Illinois, U.S.A.
John P. Oberdier = Abbott Laboratories, Abbott Park, Illinois, U.S.A.
Thomas M. O’Connell = GlaxoSmithKline, Research Triangle Park, North Carolina, U.S.A.
Claudia C. Okeke = Department of Patient Safety, United States Pharmacopeia,
Rockville, Maryland, U.S.A.
Clyde M. Ofner III = University of the Sciences in Philadelphia, Philadelphia, Pennsylvania, U.S.A.
Rosa Maria Olmo = Departamento de Bioquimica y Biologia Molecular, Universidad Complutense
de Madrid, Madrid, Spain
Wayne P. Olson = Beecher, Illinois, U.S.A.
Bridget O’Mahony = Department of Pharmaceutical Sciences, University of Strathclyde,
Glasgow, U.K.
Christine K. O’Neil = School of Pharmacy, Duquesne University, Pittsburgh, Pennsylvania, U.S.A.
Tooru Ooya = Japan Advanced Institute of Science and Technology, Ishikawa, Japan
Damon Osbourn = Department of Chemistry, University of Kansas, Lawrence, Kansas, U.S.A.
Rama V. Padmanabhan = ALZA Corporation, Spring Lake Park, Minnesota, U.S.A.
Sariputta P. Pakhale = National Institute of Pharmaceutical Education and Research,
Punjab, India
Mark G. Papich = Department of Anatomy, Physiological Sciences and Radiology, North Carolina
State University, Raleigh, North Carolina, U.S.A.
Jagdish Parasrampuria = Johnson & Johnson, Skillman, New Jersey, U.S.A.
Eun Jung Park = Department of Medicinal Chemistry and Pharmacognosy, College of Pharmacy,
University of Illinois at Chicago, Chicago, Illinois, U.S.A.
Jung Y. Park = Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, Connecticut, U.S.A.
Kinam Park = Departments of Pharmaceutics and Biomedical Engineering, Purdue University,
West Lafayette, Indiana, U.S.A.
Diane M. Paskiet = West Monarch Analytical Laboratories, Maumee, Ohio, U.S.A.
Barbara Perry = Hoffmann-La Roche, Welwyn, U.K.
Adam Persky = Department of Pharmaceutics, College of Pharmacy, University of Florida,
Gainesville, Florida, U.S.A.
Gregory F. Peters = Lab Safety Corporation, Des Plaines, Illinois, U.S.A.
S. Petteway, Jr. = Bayer Corporation, Research Triangle Park, North Carolina, U.S.A.
John M. Pezzuto = School of Pharmacy and Pharmaceutical Sciences, Purdue University,
West Lafayette, Indiana, U.S.A.
J. Bradley Phipps = ALZA Corporation, Mountain View, California, U.S.A.
D. Pifat = Bayer Corporation, Research Triangle Park, North Carolina, U.S.A.
Michael J. Pikal = School of Pharmacy, University of Connecticut, Storrs, Connecticut, U.S.A.
Wayne L. Pines = Pharmaceutical Consultant, Washington, D.C., U.S.A.
Terezinha de Jesus Andreo Pinto = Department of Pharmacy, University of Sao Paulo,
Sao Paulo, Brazil
Dario Pistolesi = Fedegari Autoclavi SpA, Albuzzano, Italy
Stephen William Pitt = Johnson & Johnson, Skillman, New Jersey, U.S.A.
Michael E. Placke = Battelle Pulmonary Therapeutics, Inc., Columbus, Ohio, U.S.A.
Fridrun Podczeck = Institute of Pharmacy, University of Sunderland, Sunderland, U.K.
Therese I. Poirier = Department of Clinical Pharmacy, Duquesne University,
Pittsburgh, Pennsylvania, U.S.A.
 xviii

Tı́mea Polgár = Gedeon Richter Ltd., Budapest, Pest, Hungary

Jacques H. Poupaert = Department of Medicinal Chemistry, Catholic University of Louvain,
Brussels, Belgium
Sunil Prabhu = College of Pharmacy, Western University of Health Sciences,
Pomona, California, U.S.A.
Neil Purdie = Department of Chemistry, Oklahoma State University, Stillwater, Oklahoma, U.S.A.
Bashir A. Qadri = Department of Pharmaceutics, Pharmacy School, The Hebrew University of
Jerusalem, Jerusalem, Israel
Fenghe Qiu = Department of Analytical Sciences, Boehringer Ingelheim Pharmaceuticals, Inc.,
Ridgefield, Connecticut, U.S.A.
Ying-shu Quan = Department of Biopharmaceutics, Kyoto Pharmaceutical University, Kyoto, Japan
Saeed A. Qureshi = Health Products and Food Branch, Health Canada, Sir F.G. Banting Research
Centre, Ottawa, Ontario, Canada
R. Raghavan = Abbott Laboratories, Abbott Park, Illinois, U.S.A.
Natarajan Rajagopalan = Lilly Research Laboratories, Eli Lilly and Company, Indianapolis,
Indiana, U.S.A.
Judy A. Raper = Department of Chemical Engineering, University of Missouri-Rolla, Rolla,
Missouri, U.S.A.
Suneel Rastogi = Forest Laboratories, Inc., Inwood, New York, U.S.A.
William R. Ravis = School of Pharmacy, Auburn University, Auburn, Alabama, U.S.A.
Brian James Reamer = SciRegs Consulting, Columbia, Maryland, U.S.A.
Robert A. Reed = Merck Research Laboratories, Merck & Co., Inc., West Point,
Pennsylvania, U.S.A.
Thomas L. Reiland = Abbott Laboratories, North Chicago, Illinois, U.S.A.
J. P. Remon = Laboratory of Pharmaceutical Technology, Ghent University, Ghent, Belgium
Steven Shijun Ren = Maxim Pharmaceuticals, Inc., San Diego, California, U.S.A.
Michael A. Repka = School of Pharmacy, The University of Mississippi, University,
Mississippi, U.S.A.
Christopher T. Rhodes = College of Pharmacy, University of Rhode Island, Kingston,
Rhode Island, U.S.A.
Martin M. Rieger = Morris Plains, New Jersey, U.S.A.
Jean G. Riess = Les Giaines, Falicon, France
Thomas N. Riley = Department of Pharmaceutical Sciences, Auburn University, Auburn,
Alabama, U.S.A.
Diane Rindgen = Schering-Plough Research Institute, Kenilworth, New Jersey, U.S.A.
Ronald J. Roberts = AstraZeneca, Cheshire, U.K.
Naı́r Rodrı́guez-Hornedo = Department of Pharmaceutical Sciences, University of Michigan,
Ann Arbor, Michigan, U.S.A.
Raymond C. Rowe = University of Bradford, Cheshire, U.K.
Alan E. Royce = Novartis Pharmaceuticals, East Hanover, New Jersey, U.S.A.
Joseph T. Rubino = Wyeth-Ayerst Research, Pearl River, New York, U.S.A.
Colleen E. Ruegger = Novartis Pharmaceuticals, East Hanover, New Jersey, U.S.A.
Effendi Rusli = Department of Chemical and Biomolecular Engineering, University of Illinois at
Urbana-Champaign, Urbana, Illinois, U.S.A.
J. Howard Rytting = Department of Pharmaceutical Chemistry, The University of Kansas,
Lawrence, Kansas, U.S.A.
Rosalie Sagraves = University of Illinois, Chicago, Illionis, U.S.A.
Dmitry Samarsky = Invitrogen Corporation, Carlsbad, California, U.S.A.
Ronald A. Sanftleben = GlaxoSmithKline, Research Triangle Park, North Carolina, U.S.A.
Sujit S. Sansgiry = Department of Clinical Sciences and Administration, College of Pharmacy,
University of Houston, Houston, Texas, U.S.A.
 xix

Peter C. Schmidt = Nuernberg, Germany

David R. Schoneker = Colorcon, Inc., West Point, Pennsylvania, U.S.A.
Stephen G. Schulman = Department of Medicinal Chemistry, University of Florida, Gainesville,
Florida, U.S.A.
Erik R. Scott = Medtronic, Inc., Brooklyn Center, Minnesota, U.S.A.
Sigrun Seeger = Schering AG, Berlin, Germany
Siri H. Segalstad = Segalstad Consulting AS, Oslo, Norway
Richard B. Seymour = Haight Ashbury Free Clinics, San Francisco, California, U.S.A.
Jaymin C. Shah = Pfizer Global Research and Development, Groton, Connecticut, U.S.A.
Umang Shah = Pfizer Global Research and Development, Morris Plains, New Jersey, U.S.A.
Utpal U. Shah = School of Pharmacy and Chemistry, Liverpool John Moores University,
Liverpool, U.K.
Shalaby W. Shalaby = Poly-Med, Inc., Anderson, South Carolina, U.S.A.
Leon Shargel = Eon Labs Manufacturing, Inc., Laurelton, New York, U.S.A.
Eric B. Sheinin = Pharmaceutical Ingredient Verification Program, United States Pharmacopeia,
Rockville, Maryland, U.S.A.
Joseph Sherma = Department of Chemistry, Lafayette College, Easton, Pennsylvania, U.S.A.
Paul J. Sheskey = The Dow Chemical Company, Midland, Michigan, U.S.A.
Troy Shinbrot = Department of Chemical and Biochemical Engineering, Rutgers University,
Piscataway, New Jersey, U.S.A.
Leonid Shnayder = Aker Kvaerner Pharmaceuticals, Bridgewater, New Jersey, U.S.A.
Gauri Shringarpure = Department of Clinical Sciences and Administration, College of Pharmacy,
University of Houston, Houston, Texas, U.S.A.
Lynn Simpson = Department of Clinical Sciences and Administration, College of Pharmacy,
University of Houston, Houston, Texas, U.S.A.
Brent D. Sinclair = Abbott Laboratories, Abbott Park, Illinois, U.S.A.
Ambarish K. Singh = Bristol-Myers Squibb Pharmaceutical Research Institute,
New Brunswick, New Jersey, U.S.A.
Brahma N. Singh = Forest Laboratories, Inc., Commack, New York, U.S.A.
Saranjit Singh = National Institute of Pharmaceutical Education and Research, Punjab, India
Shailesh K. Singh = Wyeth Research, Pearl River, New York, U.S.A.
Jerome P. Skelly = Pharmaceutical Consultant, Alexandria, Virginia, U.S.A.
Michael F. Skinner = Rhodes University, Grahamstown, South Africa
William H. Slattery III = House Ear Clinic, Los Angeles, California, U.S.A.
A. Ian Smith = Baker Heart Research Institute, Melbourne, Victoria, Australia
David E. Smith = Haight Ashbury Free Clinics, San Francisco, California, U.S.A.
Edward J. Smith = Wyeth Pharmaceuticals, Collegeville, Pennsylvania, U.S.A.
Marshall Steinberg = International Pharmaceuticals Council, Kennett Square, Pennsylvania, U.S.A.
Ralph Stone = Alcon Laboratories, Fort Worth, Texas, U.S.A.
Robert G. Strickley = Gilead Sciences Inc., Foster City, California, U.S.A.
Muppalla Sukumar = Lilly Research Laboratories, Eli Lilly and Company, Indianapolis,
Indiana, U.S.A.
Raj Suryanarayanan = College of Pharmacy, University of Minnesota,
Minneapolis, Minnesota, U.S.A.
Stuart R. Suter = Patent Attorney and Consultant, Glenside, Pennsylvania, U.S.A.
Lynda Sutton = Cato Research Ltd., Durham, North Carolina, U.S.A.
C. Jeanne Taborsky = SciRegs Consulting, Columbia, Maryland, U.S.A.
Hua Tang = TransForm Pharmaceuticals, Inc., Lexington, Massachusetts, U.S.A.
P. Tang = University of Sydney, Sydney, New South Wales, Australia
Kevin M.G. Taylor = School of Pharmacy, University of London, London, U.K.
 xx

José Marı́a Teijón = Departamento de Bioquimica y Biologia Molecular, Universidad Complutense

de Madrid, Madrid, Spain
Allen C. Templeton = Merck Research Laboratories, Merck & Co., Inc.,
West Point, Pennsylvania, U.S.A.
Bernard Testa = Institute of Medicinal Chemistry, University of Lausanne, Lausanne, Switzerland
Maya Thanou = Centre for Polymer Therapeutics, Welsh School of Pharmacy, Cardiff University,
Cardiff, U.K.
C. Thomasin = The R.W. Johnson Pharmaceutical Research Institute, Schaffhausen, Switzerland
Diane O. Thompson = CyDex, Inc., Overland Park, Kansas, U.S.A.
William J. Thomsen = Arena Pharmaceuticals, San Diego, California, U.S.A.
Youqin Tian = Alcon Laboratories, Fort Worth, Texas, U.S.A.
Jeffrey Tidwell = GlaxoSmithKline, Research Triangle Park, North Carolina, U.S.A.
Stephen Tindal = Cardinal Health Pharmaceutical Development, Somerset, New Jersey, U.S.A.
James E. Tingstad = Tingstad Associates, Green Valley, Arizona, U.S.A.
A. K. Tiwary = Department of Pharmaceutical Sciences and Drug Research, Punjabi University,
Patiala, India
Hanne Hjorth Tonnesen = School of Pharmacy, University of Oslo, Oslo, Norway
Edward H. Trappler = Lyophilization Technology, Inc., Ivyland, Pennsylvania, U.S.A.
Richard Turton = Department of Chemical Engineering, College of Engineering and Mineral
Resources, West Virginia University, Morgantown, West Virginia, U.S.A.
Mitsuru Uchiyama = Society of Japanese Pharmacopeia, Tokyo, Japan
Merlin L. Utter = Wyeth Research, Pearl River, New York, U.S.A.
Madhu K. Vadnere = Schein Pharmaceutical, Inc., Florham Park, New Jersey, U.S.A.
Stephen J. Valazza = Novartis Pharmaceuticals, East Hanover, New Jersey, U.S.A.
Lynn Van Campen = Nektar Therapeutics, San Carlos, California, U.S.A.
Koen Van Deun = Janssen Pharmaceutica N.V., Beerse, Belgium
Mark D. VanArendonk = Pharmacia Corporation, Kalamazoo, Michigan, U.S.A.
Jason M. Vaughn = College of Pharmacy, The University of Texas at Austin, Austin, Texas, U.S.A.
Christine Vauthier = School of Pharmacy, University of Paris XI, Cha^tenay-Malabry, France
Helena J. Venables = School of Pharmacy and Chemistry, Liverpool John Moores University,
Liverpool, U.K.
Geraldine Venthoye = Inhale Therapeutic Systems, Inc., San Carlos, California, U.S.A.
J. Coos Verhoef = Division of Pharmaceutical Technology, Leiden/Amsterdam Center for Drug
Research, Leiden, The Netherlands
C. Vervaet = Laboratory of Pharmaceutical Technology, Ghent University, Ghent, Belgium
Vesa Virtanen = Department of Pharmaceutical Product Development,
Orion Pharma, Kuopio, Finland
Imre M. Vitez = Bristol-Myers Squibb Pharmaceutical Research Institute,
New Brunswick, New Jersey, U.S.A.
Karel Vytras = Department of Analytical Chemistry, University of Pardubice, Pardubice,
Czech Republic
Robert F. Wagner = Novartis Pharmaceuticals, East Hanover, New Jersey, U.S.A.
Michelle Wake = Library and Information Services, School of Pharmacy, University of London,
London, U.K.
Roderick B. Walker = Rhodes University, Grahamstown, South Africa
Kenneth A. Walters = An-eX Analytical Services, Ltd., Cardiff, U.K.
Ch. Wandrey = Laboratoire des Polye´lectrolytes et BioMacromolécules,
EPFL, Lausanne, Switzerland
Richard Washkuhn = Consultant, Lexington, Kentucky, U.S.A.
Alan L. Weiner = Alcon Research, Ltd., Fort Worth, Texas, U.S.A.
 xxi

Peter Welch = Invitrogen Corporation, Carlsbad, California, U.S.A.

Peter G. Welling = Kent, U.K.
Mickey L. Wells = GlaxoSmithKline, Research Triangle Park, North Carolina, U.S.A.
Hong Wen = Wyeth Research, Pearl River, New York, U.S.A.
Albert I. Wertheimer = School of Pharmacy, Temple University, Philadelphia, Pennsylvania, U.S.A.
Paul K. Whitcraft = Rolled Alloys, Temperance, Michigan, U.S.A.
Cheryl A. Wiens = University of Alberta, Edmonton, Alberta, Canada
Nina F. Wilkins = Acrux DDS Pty Ltd., West Melbourne, Victoria, Australia
Ellen M. Williams = Pfizer, Inc., New York, New York, U.S.A.
Paul J. Williams = Department of Pharmacy Practice, School of Pharmacy, University of the Pacific,
Stockton, California, U.S.A.
Robert O. Williams III = College of Pharmacy, The University of Texas at Austin,
Austin, Texas, U.S.A.
Roger L. Williams = United States Pharmacopeia, Rockville, Maryland, U.S.A.
Clive G. Wilson = Department of Pharmaceutical Sciences, University of Strathclyde,
Glasgow, U.K.
M. G. Wing = Biosciences Division, Huntingdon Life Sciences, Cambridgeshire, U.K.
Tin Wui Wong = University Technology MARA, Selangor, Malaysia
A. Wayne Wood = GlaxoSmithKline, Research Triangle Park, North Carolina, U.S.A.
A. David Woolfson = School of Pharmacy, The Queen’s University of Belfast, Belfast, U.K.
Thomas J. Wubben = Department of Chemical and Biomolecular Engineering, University of Illinois at
Urbana-Champaign, Urbana, Illinois, U.S.A.
Dawei Xuan = Pfizer Global Research and Development, Ann Arbor, Michigan, U.S.A.
Samuel H. Yalkowsky = Department of Pharmacy Practice and Science, College of Pharmacy,
The University of Arizona, Tucson, Arizona, U.S.A.
Hiroshi Yamahara = Tanabe Seiyaku Co., Ltd., Osaka, Japan
Victor C. Yang = Department of Pharmaceutical Sciences, The University of Michigan,
Ann Arbor, Michigan, U.S.A.
Charles R. Yates = Department of Pharmaceutical Sciences, College of Pharmacy,
University of Tennessee, Memphis, Tennessee, U.S.A.
Yoon Yeo = Department of Industrial and Physical Pharmacy, Purdue University,
West Lafayette, Indiana, U.S.A.
Andrew B.C. Yu = Center for Drug Evaluation and Research, U.S. Food and Drug Administration,
Rockville, Maryland, U.S.A.
Mark J. Zellhofer = Guilford Pharmaceuticals, Inc., Baltimore, Maryland, U.S.A.
Feng Zhang = College of Pharmacy, University of Texas at Austin, Austin, Texas, U.S.A.
William C. Zimlich, Jr. = Battelle Pulmonary Therapeutics, Inc., Columbus, Ohio, U.S.A.
Contents

Contributors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ix
Topical Table of Contents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xxxi
Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . liii

Volume 1
21 CFR Part 11 Revisited = Thomas Linz and Sigrun Seeger . . . . . . . . . . . . . . . . . . . . . . . . 1
Absorption Enhancers = J. P. Remon . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Absorption of Drugs = Peter G. Welling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
Adsorption at Solid Surfaces: Pharmaceutical Applications = Hong Wen . . . . . . . . . . . . . . . . . 34
Adverse Drug Reactions = Therese I. Poirier and Robert L. Maher, Jr. . . . . . . . . . . . . . . . . . . 46
Advertising and Promotion of Prescription and Over-the-Counter Drug Products =
Wayne L. Pines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
Alternative Medicines = Kristi L. Lenz . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66
Amorphous Pharmaceutical Systems = Bruno C. Hancock . . . . . . . . . . . . . . . . . . . . . . . . . . 83
Analytical Procedures: Validation = Joachim Ermer and John S. Landy . . . . . . . . . . . . . . . . . 92
Animals in Drug Development = Farrel L. Fort . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114
Aseptic Processing: Validation = James P. Agalloco and James E. Akers . . . . . . . . . . . . . . . . 127
Autoxidation and Antioxidants = John M. Pezzuto and Eun Jung Park . . . . . . . . . . . . . . . . . . 139
Bioabsorbable Polymers = Shalaby W. Shalaby . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155
Bioavailability of Drugs and Bioequivalence = James T. Dalton and Charles R. Yates . . . . . . . . 164
Biodegradable Polymers as Drug Carriers = Peter Markland and Victor C. Yang . . . . . . . . . . . 176
Biologic Fluids: Analysis = Stephen G. Schulman, Judith A. Davis, and Gayle A. Brazeau . . . . . 194
Biopharmaceutics = Leon Shargel and Andrew B.C. Yu . . . . . . . . . . . . . . . . . . . . . . . . . . . 208
Biosynthesis of Drugs = Geoffrey A. Cordell and Kyung Hee Lee . . . . . . . . . . . . . . . . . . . . . 228
Biotechnology and Biological Preparations = Ronald P. Evens . . . . . . . . . . . . . . . . . . . . . . . 258
Biotechnology-Derived Drug Products: Formulation Development =
Mary E.M. Cromwell . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 281
Biotechnology-Derived Drug Products: Stability Testing, Filling, and Packaging =
Mary E.M. Cromwell . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 302
Biotransformation of Drugs = Les F. Chasseaud and D. R. Hawkins . . . . . . . . . . . . . . . . . . . 310
Bio-Validation of Steam Sterilization = Nigel A. Halls . . . . . . . . . . . . . . . . . . . . . . . . . . . . 325
Blood Substitutes: Fluorocarbon Approach = Jean G. Riess . . . . . . . . . . . . . . . . . . . . . . . . . 335
Blood Substitutes: Hemoglobin-Based Oxygen Carriers = Deanna J. Nelson . . . . . . . . . . . . . . . 353
Blow-Fill-Seal: Advanced Aseptic Processing = Deborah J. Jones . . . . . . . . . . . . . . . . . . . . . . 378
Buffers, Buffering Agents, and Ionic Equilibria = Harry G. Brittain . . . . . . . . . . . . . . . . . . . . 385
Calorimetry in Pharmaceutical Research and Development = Sophie-Dorothe´e Clas,
Chad R. Dalton, and Bruno C. Hancock . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 393
Capsules, Hard = Brian E. Jones . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 406
Capsules, Soft = David H. Bergstrom, Stephen Tindal, and Wenbin Dang . . . . . . . . . . . . . . . 419
Carcinogenicity Testing: Past, Present, and Future = Koen Van Deun . . . . . . . . . . . . . . . . . . . 431

xxiii
 xxiv

Chiroptical Analytical Methods = Neil Purdie . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 445

Chromatographic Methods of Analysis: Gas Chromatography = Isadore Kanfer,
Roderick B. Walker, and Michael F. Skinner . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 463
Chromatographic Methods of Analysis: High Performance Liquid Chromatography =
R. Raghavan and Jose C. Joseph . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 526
Chromatographic Methods of Analysis: Thin Layer Chromatography = Joseph Sherma . . . . . . . . 538
Clinical Data Management Systems = Samuel V. Givens, Debra Barnes,
Victoria Imber, and Barbara Perry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 551
Clinical Evaluation of Drugs = Lynda Sutton, Allen Cato, and Allen Cato III . . . . . . . . . . . . . 560
Clinical Pharmacokinetics and Pharmacodynamics = Leslie Z. Benet and
Laviero Mancinelli . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 572
Clinical Supplies Manufacture: GMP Considerations = David L. Chesney and
Mark L. Balboni . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 591
Coacervation and Phase Separation = Bruno Gander, Maria Jose Blanco-Prı´eto,
C. Thomasin, Ch. Wandrey, and D. Hunkeler . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 600
Cocrystals: Design, Properties and Formation Mechanisms = Naı´r Rodrı´guez-Hornedo,
Sarah J. Nehm, and Adivaraha Jayasankar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 615
Colloids and Colloid Drug Delivery System = Diane J. Burgess . . . . . . . . . . . . . . . . . . . . . . . 636
Coloring Agents for Use in Pharmaceuticals = David R. Schoneker . . . . . . . . . . . . . . . . . . . . 648

Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . I1

Volume 2
Complexation: Cyclodextrins = Gerold Mosher and Diane O. Thompson . . . . . . . . . . . . . . . . 671
Complexation: Non-Cyclodextrins = Galina N. Kalinkova . . . . . . . . . . . . . . . . . . . . . . . . . . 697
Computer Systems Validation = Orlando Lopez . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 707
Computer-Assisted Drug Design = J. Phillip Bowen and Michael Cory . . . . . . . . . . . . . . . . . . 714
Computers in Pharmaceutical Technology = Onkaram Basavapathruni . . . . . . . . . . . . . . . . . . 732
Continuous Processing of Pharmaceuticals = J. P. Remon and C. Vervaet . . . . . . . . . . . . . . . . 743
Contract Manufacturing = Duncan E. McVean . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 750
Cooling Processes and Congealing = Richard Turton and Xiu Xiu Cheng . . . . . . . . . . . . . . . . 761
Coprecipitates and Melts = Madhu K. Vadnere . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 774
Corrosion of Pharmaceutical Equipment = Paul K. Whitcraft . . . . . . . . . . . . . . . . . . . . . . . . 782
Cosmetics and Their Relation to Drugs = Martin M. Rieger . . . . . . . . . . . . . . . . . . . . . . . . . 798
Cosolvents and Cosolvency = Joseph T. Rubino . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 806
Crystal Habit Changes and Dosage Form Performance = A. K. Tiwary . . . . . . . . . . . . . . . . . . 820
Crystallization: General Principles and Significance on Product Development =
Naı´r Rodrı´guez-Hornedo, Ron C. Kelly, Brent D. Sinclair, and Jonathan M. Miller . . . . . . . 834
Crystallization: Particle Size Control = Richard D. Braatz, Mitsuko Fujiwara,
Thomas J. Wubben, and Effendi Rusli . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 858
Dendrimers = Antony D’Emanuele, David Attwood, and Ragheb Abu-Rmaileh . . . . . . . . . . . . 872
Dental Products = Sebastian G. Ciancio . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 891
Dissolution and Dissolution Testing = A. Mark Dyas and Utpal U. Shah . . . . . . . . . . . . . . . . . 908
DNA Probes for the Identification of Microbes = Wayne P. Olson . . . . . . . . . . . . . . . . . . . . . 929
Dosage Form Design: A Physicochemical Approach = Michael B. Maurin and
Anwar A. Hussain . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 939
Dosage Forms and Basic Preparations: History = Robert A. Buerki and Gregory J. Higby . . . . . . 948
Dosage Forms: Lipid Excipients = Alan L. Weiner . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 975
Dosage Forms: Non-Parenterals = Shailesh K. Singh and Venkatesh Naini . . . . . . . . . . . . . . . 988
Dosage Forms: Parenterals = Gayle A. Brazeau, Adam Persky, and Jintana Napaporn . . . . . . . 1001
 xxv

Dosage Regimens and Dose-Response = Chyung S. Cook and Aziz Karim . . . . . . . . . . . . . . . . 1012
Dressings in Wound Management = Sarah M.E. Cockbill . . . . . . . . . . . . . . . . . . . . . . . . . . 1023
Drug Abuse = Richard B. Seymour and David E. Smith . . . . . . . . . . . . . . . . . . . . . . . . . . . 1038
Drug Delivery Systems: Neutron Scattering Studies = M. Jayne Lawrence and
David J. Barlow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1049
Drug Delivery: Buccal Route = James C. McElnay and Carmel M. Hughes . . . . . . . . . . . . . . . 1071
Drug Delivery: Controlled Release = Yie W. Chien and Senshang Lin . . . . . . . . . . . . . . . . . . . 1082
Drug Delivery: Fast-Dissolve Systems = Vikas Agarwal, Bhavesh H. Kothari,
Derek V. Moe, and Rajendra K. Khankari . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1104
Drug Delivery: Liquid Crystals in = Christel C. Mueller-Goymann . . . . . . . . . . . . . . . . . . . . 1115
Drug Delivery: Monoclonal Antibodies = John B. Cannon, Ho-Wah Hui, and
Pramod K. Gupta . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1132
Drug Delivery: Monoclonal Antibodies in Imaging and Therapy = Ban-An Khaw . . . . . . . . . . . . 1149
Drug Delivery: Mucoadhesive Hydrogels = Hans E. Junginger, J. Coos Verhoef, and
Maya Thanou . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1169
Drug Delivery: Nanoparticles = Elias Fattal and Christine Vauthier . . . . . . . . . . . . . . . . . . . 1183
Drug Delivery: Nasal Route = Rene´ Bommer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1201
Drug Delivery: Needle-Free Systems = Toby King . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1209
Drug Delivery: Ophthalmic Route = Masood Chowhan, Alan L. Weiner, and
Haresh Bhagat . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1220
Drug Delivery: Oral Colon-Specific = Vincent H.L. Lee and Suman K. Mukherjee . . . . . . . . . . 1228
Drug Delivery: Oral Route = Brahma N. Singh and Kwon H. Kim . . . . . . . . . . . . . . . . . . . . 1242
Drug Delivery: Parenteral Route = Michael J. Akers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1266
Drug Delivery: Pulmonary Delivery = Michael T. Newhouse . . . . . . . . . . . . . . . . . . . . . . . . . 1279
Drug Delivery: Pulsatile Systems = Till Bussemer and Roland A. Bodmeier . . . . . . . . . . . . . . . 1287
Drug Delivery: Rectal Route = J. Howard Rytting and Joseph A. Fix . . . . . . . . . . . . . . . . . . . 1298
Drug Delivery: Topical and Transdermal Routes = Kenneth A. Walters . . . . . . . . . . . . . . . . . . 1311
Drug Delivery: Tumor-Targeted Systems = Yu Li and Chao-Pin Lee . . . . . . . . . . . . . . . . . . . 1326
Drug Delivery: Vaginal Route = Yie W. Chien and Chi H. Lee . . . . . . . . . . . . . . . . . . . . . . . 1339
Drug Design: Basic Principles and Applications = Jacques H. Poupaert . . . . . . . . . . . . . . . . . . 1362
Drug Development Management = James E. Tingstad . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1370
Drug Information Systems = Linda Lisgarten and Michelle Wake . . . . . . . . . . . . . . . . . . . . . 1385
Drug Interactions = Daniel A. Hussar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1392
Drug Master Files = C. Jeanne Taborsky and Brian James Reamer . . . . . . . . . . . . . . . . . . . 1401
Drug Safety Evaluation = Farrel L. Fort . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1406
Dry Powder Aerosols: Emerging Technologies = Hak-Kim Chan and Nora Y.K. Chew . . . . . . . . 1428
Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . I1

Volume 3
Drying and Dryers = Anthony J. Hlinak and Bradley A. Clark . . . . . . . . . . . . . . . . . . . . . . 1435
Economic Characteristics of the R&D Intensive Pharmaceutical Industry = Douglas L. Cocks . . . . 1450
Effervescent Pharmaceuticals = Nils-Olof Lindberg and Henri Hansson . . . . . . . . . . . . . . . . . 1454
Elastomeric Components for the Pharmaceutical Industry = Edward J. Smith . . . . . . . . . . . . . . 1466
Electrical Power Systems for Pharmaceutical Equipment = Joseph Maida . . . . . . . . . . . . . . . . 1482
Electroanalytical Methods of Analysis: Polarography and Voltammetry =
A. David Woolfson . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1490
Electroanalytical Methods of Analysis: Potentiometry = Karel Vytras . . . . . . . . . . . . . . . . . . . 1502
Electrochemical Detection for Pharmaceutical Analysis = Craig E. Lunte and
Damon Osbourn . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1516
 xxvi

Electrostatic Charge in Pharmaceutical Systems = Philip Chi Lip Kwok and

Hak-Kim Chan . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1535
Emulsions and Microemulsions = Gillian M. Eccleston . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1548
Enzyme Immunoassay and Related Bioanalytical Methods = John W.A. Findlay and
Ira Das . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1566
Equipment Cleaning = Destin A. LeBlanc . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1580
European Agency for the Evaluation of Medicinal Products (EMEA) = David M. Jacobs . . . . . . 1593
Evaporation and Evaporators = David P. Kessler . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1600
Excipients for Pharmaceutical Dosage Forms = Patrick J. Crowley and Luigi G. Martini . . . . . . 1609
Excipients: Parenteral Dosage Forms and Their Role = Sandeep Nema,
Ron J. Brendel, and Richard Washkuhn . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1622
Excipients: Powders and Solid Dosage Forms = Hak-Kim Chan and Nora Y.K. Chew . . . . . . . . 1646
Excipients: Safety Testing = Marshall Steinberg and Florence K. Kinoshita . . . . . . . . . . . . . . 1656
Expert Systems in Pharmaceutical Product Development =
Raymond C. Rowe and Ronald J. Roberts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1663
Expiration Dating = Leslie C. Hawley and Mark D. VanArendonk . . . . . . . . . . . . . . . . . . . . 1685
Extractables and Leachables in Drugs and Packaging = Daniel L. Norwood,
Alice T. Granger, and Diane M. Paskiet . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1693
Extrusion and Extruders = J. M. Newton . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1712
Film Coating of Oral Solid Dosage Forms = Linda A. Felton . . . . . . . . . . . . . . . . . . . . . . . . 1729
Filters and Filtration = Maik W. Jornitz . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1748
Flame Photometry = Thomas M. Nowak . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1759
Flavors and Flavor Modifiers = Thomas L. Reiland and John M. Lipari . . . . . . . . . . . . . . . . . 1763
Fluid Bed Processes for Forming Functional Particles =
Yoshinobu Fukumori and Hideki Ichikawa . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1773
Food and Drug Administration: Role in Drug Regulation = Roger L. Williams . . . . . . . . . . . . . 1779
Fractal Geometry in Pharmaceutical and Biological Applications = P. Tang,
Hak-Kim Chan, and Judy A. Raper . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1791
Freeze Drying = Michael J. Pikal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1807
Freeze Drying, Scale-Up Considerations = Edward H. Trappler . . . . . . . . . . . . . . . . . . . . . . 1834
Gastro-Retentive Systems = Amnon Hoffman and Bashir A. Qadri . . . . . . . . . . . . . . . . . . . 1850
Gelatin-Containing Formulations: Changes in Dissolution Characteristics =
Saranjit Singh and Sariputta P. Pakhale . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1861
Gels and Jellies = Clyde M. Ofner III and Cathy M. Klech-Gelotte . . . . . . . . . . . . . . . . . . . 1875
Generic Drugs and Generic Equivalency = Arthur H. Kibbe . . . . . . . . . . . . . . . . . . . . . . . . . 1891
Genetic Aspects of Drug Development = Werner Kalow . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1897
Geriatric Dosing and Dosage Forms = Cheryl A. Wiens and Carol A. Borynec . . . . . . . . . . . . . 1905
Good Clinical Practices (GCPs): An Overview = Richard A. Guarino . . . . . . . . . . . . . . . . . . . 1925
Good Laboratory Practices (GLPs): An Overview = Nigel J. Dent . . . . . . . . . . . . . . . . . . . . . 1931
Good Manufacturing Practices (GMPs): An Overview = Ira R. Berry . . . . . . . . . . . . . . . . . . . 1941
Handling Hazardous Chemicals and Pharmaceuticals = Antonio Conto . . . . . . . . . . . . . . . . . . 1948
Harmonization of Pharmacopeial Standards = Lee T. Grady and Jerome A. Halperin . . . . . . . . 1955
Headspace Oxygen Analysis in Pharmaceutical Products = Allen C. Templeton and
Robert A. Reed . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1967
Health Care Systems: Outside the United States = Albert I. Wertheimer and
Sheldon X. Kong . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1977
Health Care Systems: Within the United States = Henri R. Manasse, Jr. . . . . . . . . . . . . . . . . . 1985
Homogenization and Homogenizers = Shailesh K. Singh and Venkatesh Naini . . . . . . . . . . . . . 1996
Hot-Melt Extrusion Technology = Jim W. McGinity, Michael A. Repka,
John J. Koleng, Jr., and Feng Zhang . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2004
 xxvii

Hydrogels = Marı´a Dolores Blanco, Rosa Maria Olmo, and José Marı´a Teijón . . . . . . . . . . . . 2021
Hydrolysis of Drugs = Jason M. LePree and Kenneth A. Connors . . . . . . . . . . . . . . . . . . . . . 2040
Immunoassay = Stephen G. Schulman and G.J.P.J. Beernink . . . . . . . . . . . . . . . . . . . . . . . . 2048
In Vitro–In Vivo Correlation = J.-M. Cardot and Erick Beyssac . . . . . . . . . . . . . . . . . . . . . . 2062
Inhalation: Dry Powder = Lynn Van Campen and Geraldine Venthoye . . . . . . . . . . . . . . . . . 2077
Inhalation: Liquids = Michael E. Placke, Jeffrey Ding, and William C. Zimlich, Jr. . . . . . . . . . 2092
Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . I1

Volume 4
Iontophoresis = J. Bradley Phipps, Erik R. Scott, J. Richard Gyory, and
Rama V. Padmanabhan . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2119
Isolators for Pharmaceutical Application = Gordon J. Farquharson . . . . . . . . . . . . . . . . . . . . 2133
Isomerism = Thomas N. Riley, Jack DeRuiter, William R. Ravis, and C. Randall Clark . . . . . . 2142
Laboratory Information Management System (LIMS) = Siri H. Segalstad . . . . . . . . . . . . . . . . 2164
Laminar Airflow Equipment: Applications and Operation = Gregory F. Peters . . . . . . . . . . . . . . 2171
Lead Optimization in Pharmaceutical Development: Molecular and Cellular Approaches =
C. K. Atterwill and M. G. Wing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2192
Lens Care Products = Masood Chowhan and Ralph Stone . . . . . . . . . . . . . . . . . . . . . . . . . 2202
Liquid Oral Preparations = Jagdish Parasrampuria and Stephen William Pitt . . . . . . . . . . . . . 2216
Lozenges = Robert W. Mendes and Hridaya Bhargava . . . . . . . . . . . . . . . . . . . . . . . . . . . 2231
Materials of Construction for Pharmaceutical Equipment = Michelle M. Gonzalez . . . . . . . . . . . 2237
Medication Errors = Diane D. Cousins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2243
Melt Processes for Oral Solid Dosage Forms = Paul W.S. Heng and Tin Wui Wong . . . . . . . . . . 2257
Metabolite Identification in Drug Discovery = Kathleen A. Cox, Nigel Clarke, and
Diane Rindgen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2262
Metered Dose Inhalers = Sandy J.M. Munro and Alan L. Cripps . . . . . . . . . . . . . . . . . . . . . 2269
Microbial Control of Pharmaceuticals = Nigel A. Halls . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2286
Microbiologic Monitoring of Controlled Processes = Gregory F. Peters and
Marghi R. McKeon . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2298
Microencapsulation Technology = Kinam Park and Yoon Yeo . . . . . . . . . . . . . . . . . . . . . . . . 2315
Microsphere Technology and Applications = Diane J. Burgess and
Anthony J. Hickey . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2328
Milling of Active Pharmaceutical Ingredients = Elizabeth S. Fisher . . . . . . . . . . . . . . . . . . . . 2339
Mixing and Segregation in Tumbling Blenders = Troy Shinbrot and
Fernando J. Muzzio . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2352
Moisture in Pharmaceutical Products = R. Gary Hollenbeck . . . . . . . . . . . . . . . . . . . . . . . . 2368
Nanoparticle Engineering = Robert O. Williams III and Jason M. Vaughn . . . . . . . . . . . . . . . . 2384
Neural Computing and Formulation Optimization = Elizabeth Colbourn and
Raymond C. Rowe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2399
Non-Prescription Drugs = Sujit S. Sansgiry, Lynn Simpson, Gauri Shringarpure, and
Amit Kulkarni . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2413
Nutraceutical Supplements = G. Brian Lockwood . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2431
Optimization Methods = Gareth A. Lewis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2452
Orphan Drugs = Carolyn H. Asbury . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2468
Otic Preparations = William H. Slattery III . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2475
Outsourcing = Duane B. Lakings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2486
Packaging Materials: Glass = Claudia C. Okeke and Desmond G. Hunt . . . . . . . . . . . . . . . . . 2508
Packaging Systems: Compendial Requirements = Claudia C. Okeke, Desmond G. Hunt,
Nicholas Mohr, Thomas Medwick, Eric B. Sheinin, and Roger L. Williams . . . . . . . . . . . . 2526
 xxviii

Paperless Documentation Systems = Ellen M. Williams and Michael McKenna . . . . . . . . . . . . 2551

Particle Engineering = Miriam K. Franchini . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2567
Particle-Size Characterization = Brian H. Kaye . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2582
Partition Coefficients = Eric J. Lien and Steven Shijun Ren . . . . . . . . . . . . . . . . . . . . . . . . 2595
Patents: International Perspective = Stuart R. Suter and Peter J. Giddings . . . . . . . . . . . . . . . 2604
Patents: United States Perspective = Lorie Ann Morgan and Jeffrey Tidwell . . . . . . . . . . . . . 2616
Pediatric Dosing and Dosage Forms = Rosalie Sagraves . . . . . . . . . . . . . . . . . . . . . . . . . . . 2629
Pelletization Techniques = Isaac Ghebre-Sellassie and Axel Knoch . . . . . . . . . . . . . . . . . . . . 2651
Peptides and Proteins: Buccal Absorption = Ashim K. Mitra, Hemant H. Alur, and
Thomas P. Johnston . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2664
Peptides and Proteins: Nasal Absorption = Takahiro Morita and Hiroshi Yamahara . . . . . . . . . 2678
Peptides and Proteins: Non-Invasive Delivery = Michael R. DeFelippis,
Muppalla Sukumar, and Natarajan Rajagopalan . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2692
Peptides and Proteins: Oral Absorption = Eugene J. McNally and Jung Y. Park . . . . . . . . . . . . 2713
Peptides and Proteins: Pulmonary Absorption = Igor Gonda . . . . . . . . . . . . . . . . . . . . . . . . . 2731
Peptides and Proteins: Transdermal Absorption = Richard H. Guy,
M. Begon ~a Delgado-Charro, and Diego Marro . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2741
Pharmaceutical Data: Mathematical Modeling = David W.A. Bourne . . . . . . . . . . . . . . . . . . . 2757
Pharmaceutical Excipient Testing: Regulatory and Preclinical Perspective = Paul Baldrick . . . . . 2771
Pharmaceutical Quality Assurance Microbiology Laboratories = Anthony M. Cundell . . . . . . . . 2783
Pharmacogenomics and Genomic Technologies = Daniel A. Brazeau . . . . . . . . . . . . . . . . . . . 2794
Pharmacokinetic/Pharmacodynamic Modeling and Simulations in Drug Development =
Dawei Xuan and Sally Y. Choe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2802
Pharmacokinetics: Effects of Food and Fasting = Rajesh Krishna and Bradford K. Jensen . . . . . 2816
Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . I1

Volume 5
Pharmacopeial Standards: European Pharmacopeia = Agnès Artiges . . . . . . . . . . . . . . . . . . . . 2829
Pharmacopeial Standards: Japanese Pharmacopeia = Mitsuru Uchiyama . . . . . . . . . . . . . . . . . 2836
Pharmacopeial Standards: United States Pharmacopeia and National Formulary =
Lee T. Grady . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2841
Photodecomposition of Drugs = Hanne Hjorth Tonnesen . . . . . . . . . . . . . . . . . . . . . . . . . . . 2859
Physiological Factors Affecting Oral Drug Delivery = Clive G. Wilson,
Bridget O’Mahony, and Blythe Lindsay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2866
Pilot Plant Design = Mickey L. Wells, Samuel B. Balik, Ralph B. Caricofe,
Charles W. Crew, Ronald A. Sanftleben, and A. Wayne Wood . . . . . . . . . . . . . . . . . . . . 2875
Pilot Plant Operation = Mickey L. Wells, A. Wayne Wood, Samuel B. Balik,
Ralph B. Caricofe, Ronald A. Sanftleben, and Charles W. Crew . . . . . . . . . . . . . . . . . . . 2886
Plants as Drugs = Christine K. O’Neil and Charles W. Fetrow . . . . . . . . . . . . . . . . . . . . . . . 2901
Polymeric Delivery Systems for Poorly Soluble Drugs = Kang Moo Huh, Sang Cheon Lee,
Tooru Ooya, and Kinam Park . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2913
Polymers in Transdermal Delivery Systems = Fumio Kamiyama and Ying-shu Quan . . . . . . . . . 2925
Polymorphism: Pharmaceutical Aspects = Harry G. Brittain . . . . . . . . . . . . . . . . . . . . . . . . . 2935
Population Pharmacokinetics = Ene I. Ette, Alaa M. Ahmad, and Paul J. Williams . . . . . . . . . 2946
Powder Sampling = Helena J. Venables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2959
Powders as Dosage Forms = Jean-Marc Aiache and Erick Beyssac . . . . . . . . . . . . . . . . . . . . 2971
Preservation of Pharmaceutical Products = Peter Gilbert and David G. Allison . . . . . . . . . . . . 2983
Process Chemistry in the Pharmaceutical Industry = Kumar G. Gadamasetti and
Ambarish K. Singh . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2993
Prodrug Design = Bernard Testa and Joachim Mayer . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3008
 xxix

Project Management = Jerome J. Groen and Cara R. Frosch . . . . . . . . . . . . . . . . . . . . . . . . 3015

Protein Binding of Drugs = Sylvie Laganière and Iain J. McGilveray . . . . . . . . . . . . . . . . . . 3027
Proteomics: Pharmaceutical Applications = A. Ian Smith . . . . . . . . . . . . . . . . . . . . . . . . . . . 3041
Pyrogens and Endotoxin Detection = James F. Cooper . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3052
Quality Assurance of Pharmaceuticals = Barbara Immel . . . . . . . . . . . . . . . . . . . . . . . . . . . 3064
Quality Systems Management = Gary C. Harbour and Robert G. Kieffer . . . . . . . . . . . . . . . . 3075
Radiochemical Methods of Analysis = R. Raghavan and Jose C. Joseph . . . . . . . . . . . . . . . . . 3082
Radiolabeling of Pharmaceutical Aerosols and Gamma Scintigraphic Imaging for
Lung Deposition = Hak-Kim Chan, Stefan Eberl, and William Glover . . . . . . . . . . . . . . 3094
Receptors for Drugs: Discovery in the Post-Genomic Era = Jeffrey M. Herz and
William J. Thomsen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3108
Rheology of Pharmaceutical Systems = Fridrun Podczeck . . . . . . . . . . . . . . . . . . . . . . . . . . 3128
RNAi in Drug Development = Dmitry Samarsky, Shelley Hough, Adam Harris,
Eugene Carstea, and Peter Welch . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3147
Roller Compaction Technology for the Pharmaceutical Industry = Ronald W. Miller and
Paul J. Sheskey . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3159
Salt Forms: Pharmaceutical Aspects = Owen I. Corrigan . . . . . . . . . . . . . . . . . . . . . . . . . . . 3177
Scale-Up and Post Approval Changes (SUPAC) = Jerome P. Skelly . . . . . . . . . . . . . . . . . . . . 3188
Scale-Up of Solid Dosage Forms = Colleen E. Ruegger, Alan E. Royce, Matthew J. Mollan, Jr.,
Robert F. Wagner, Stephen J. Valazza, and Mark R. Mecadon . . . . . . . . . . . . . . . . . . . . 3193
Secondary Electron Microscopy in Pharmaceutical Technology = Peter C. Schmidt . . . . . . . . . . 3217
Semisolid Preparations = Guru Betageri and Sunil Prabhu . . . . . . . . . . . . . . . . . . . . . . . . . 3257
Solids: Flow Properties = Stephen A. Howard . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3275
Solid-State NMR in the Characterization of Pharmaceutical Formulations =
Eric J. Munson and Joseph W. Lubach . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3297
Solubilization of Drugs in Aqueous Media = Paul B. Myrdal and Samuel H. Yalkowsky . . . . . . . 3311
Solubilizing Excipients in Pharmaceutical Formulations = Robert G. Strickley . . . . . . . . . . . . . . 3334
Spectroscopic Methods of Analysis: Atomic Absorption and Emission Spectrophotometry =
John P. Oberdier . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3367
Spectroscopic Methods of Analysis: Diffuse Reflectance Spectroscopy =
Herbert Michael Heise . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3375
Spectroscopic Methods of Analysis: Fluorescence Spectroscopy = Stephen G. Schulman and
Jeffrey A. Hughes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3387
Spectroscopic Methods of Analysis: Infrared Spectroscopy = Marilyn D. Duerst . . . . . . . . . . . . 3405
Spectroscopic Methods of Analysis: Mass Spectrometry = Mike S. Lee . . . . . . . . . . . . . . . . . . 3419
Spectroscopic Methods of Analysis: Near-Infrared Spectrometry = Emil W. Ciurczak . . . . . . . . . 3434
Spectroscopic Methods of Analysis: Nuclear Magnetic Resonance Spectroscopy =
Thomas M. O’Connell and Kevin L. Facchine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3440
Spectroscopic Methods of Analysis: Ultraviolet and Visible Spectrophotometry =
R. Raghavan and Jose C. Joseph . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3460
Starches and Starch Derivatives = Ann W. Newman, Ronald L. Mueller,
Imre M. Vitez, and Chris C. Kiesnowski . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3476
Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . I1

Volume 6
Statistical Methods = Charles Bon . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3483
Statistical Process Control and Process Capability = Thomas D. Murphy,
Shailesh K. Singh, and Merlin L. Utter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3499
Sterilization: Dry Heat = Andrea Chieppo and Thomas Kupiec . . . . . . . . . . . . . . . . . . . . . . 3512
Sterilization: Ethylene Oxide = Terezinha de Jesus Andreo Pinto . . . . . . . . . . . . . . . . . . . . . 3519
 xxx

Sterilization: Moist Heat = Dario Pistolesi . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3529

Sterilization: Radiation = Stephen G. Schulman and Phillip M. Achey . . . . . . . . . . . . . . . . . . 3540
Super Disintegrants: Characterization and Function = Larry L. Augsburger,
Albert W. Brzeczko, Umang Shah, and Huijeong Ashley Hahm . . . . . . . . . . . . . . . . . . . . 3553
Supercritical Fluid Technology in Pharmaceutical Research = Aaron S. Mayo and
Uday B. Kompella . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3568
Surfactants in Pharmaceutical Products and Systems = Owen I. Corrigan and
Anne Marie Healy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3583
Suspensions = Robert A. Nash . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3597
Tablet Compression: Machine Theory, Design, and Process Troubleshooting =
Michael J. Bogda . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3611
Tablet Evaluation Using Near-Infrared Spectroscopy = Christopher T. Rhodes and
Karen Morisseau . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3630
Tablet Formulation = Larry L. Augsburger and Mark J. Zellhofer . . . . . . . . . . . . . . . . . . . . 3641
Tablet Manufacture = Norman Anthony Armstrong . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3653
Tablet Manufacture by Direct Compression = Norman Anthony Armstrong . . . . . . . . . . . . . . . 3673
Tablet Press Instrumentation = Michael Levin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3684
Tablet Testing = Saeed A. Qureshi . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3707
Technology Transfer Considerations for Pharmaceuticals = Ira R. Berry . . . . . . . . . . . . . . . . . 3717
Thermal Analysis of Drugs and Drug Products = Danie`lle Giron . . . . . . . . . . . . . . . . . . . . . . 3726
Titrimetry = Vesa Virtanen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3752
Tonicity = Jaymin C. Shah . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3768
Tooling for Tableting = Annette Bauer-Brandl . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3782
Trace Level Impurity Analysis = Daniel L. Norwood, Fenghe Qiu, and James O. Mullis . . . . . . . 3797
Transdermal Delivery: Anatomical Site Influence = Nora Y.K. Chew,
Nina F. Wilkins, and Barrie C. Finnin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3814
Transdermal Delivery: Sonophoresis = Samir S. Mitragotri, Hua Tang,
E. Daniel Blankschtein, and Robert Langer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3828
Transdermal Delivery: Technologies = S. Kevin Li . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3843
Ultrasonic Nebulizers = Kevin M.G. Taylor and Orla McCallion . . . . . . . . . . . . . . . . . . . . . 3854
Unit Processes in Pharmacy: Fundamentals = Anthony J. Hickey and David Ganderton . . . . . . . 3862
Unit Processes in Pharmacy: Operations = Anthony J. Hickey and David Ganderton . . . . . . . . 3879
Vaccines and Other Immunological Products = Suresh K. Mittal, Harm HogenEsch, and
Kinam Park . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3908
Validation of Pharmaceutical Processes = Robert A. Nash . . . . . . . . . . . . . . . . . . . . . . . . . . 3928
Veterinary Dosage Forms = J. Desmond Baggot . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3941
Veterinary Pharmaceuticals: Factors Influencing Their Development and Use =
Marilyn N. Martinez, Laura Hungerford, and Mark G. Papich . . . . . . . . . . . . . . . . . . . . 3978
Viral Inactivation Issues in Aseptically Processed Parenterals = J. Hotta, A. Klos,
S. Petteway, Jr., and D. Pifat . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3997
Virtual Screening = Tı´mea Polgár and György M. Keserü . . . . . . . . . . . . . . . . . . . . . . . . . . 4013
Water for Pharmaceuticals = Leonid Shnayder . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4039
Water Sorption of Drugs and Dosage Forms = Mark J. Kontny and James J. Conners . . . . . . . . 4049
Waxes = Roland A. Bodmeier . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4066
Wet Granulation: End-Point Determination and Scale-Up = Michael Levin . . . . . . . . . . . . . . . 4078
World Health Organization (WHO): Global Harmonization of Requirements for
Medicinal Products = Juhana E. Idanpaan-Heikkila . . . . . . . . . . . . . . . . . . . . . . . . . . 4099
X-Ray Powder Diffractometry = Raj Suryanarayanan and Suneel Rastogi . . . . . . . . . . . . . . . 4103
Zeta Potential = Luk Chiu Li and Youqin Tian . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4117
Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . I1
Topical Table of Contents

Analytical Methods

Chromatographic Methods
Chromatographic Methods of Analysis: Gas Chromatography = Isadore Kanfer,
Roderick B. Walker, and Michael F. Skinner . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 463
Chromatographic Methods of Analysis: High Performance Liquid Chromatography =
R. Raghavan and Jose C. Joseph . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 526
Chromatographic Methods of Analysis: Thin Layer Chromatography = Joseph Sherma . . . . . . . . 538

Electroanalytical Methods
Electroanalytical Methods of Analysis: Polarography and Voltammetry =
A. David Woolfson . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1490
Electroanalytical Methods of Analysis: Potentiometry = Karel Vytras . . . . . . . . . . . . . . . . . . . 1502
Electrochemical Detection for Pharmaceutical Analysis = Craig E. Lunte and
Damon Osbourn . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1516

Miscellaneous Methods
Analytical Procedures: Validation = Joachim Ermer and John S. Landy . . . . . . . . . . . . . . . . . 92
Biologic Fluids: Analysis = Stephen G. Schulman, Judith A. Davis, and
Gayle A. Brazeau . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 194
Chiroptical Analytical Methods = Neil Purdie . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 445
Good Laboratory Practices (GLPs): An Overview = Nigel J. Dent . . . . . . . . . . . . . . . . . . . . . 1931
Headspace Oxygen Analysis in Pharmaceutical Products = Allen C. Templeton and
Robert A. Reed . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1967
Drug Delivery Systems: Neutron Scattering Studies = M. Jayne Lawrence and
David J. Barlow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1049
Laboratory Information Management System (LIMS) = Siri H. Segalstad . . . . . . . . . . . . . . . . 2164
Metabolite Identification in Drug Discovery = Kathleen A. Cox, Nigel Clarke, and
Diane Rindgen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2262
Titrimetry = Vesa Virtanen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3752
Trace Level Impurity Analysis = Daniel L. Norwood, Fenghe Qiu, and James O. Mullis . . . . . . . 3797

Radiochemical Methods
Enzyme Immunoassay and Related Bioanalytical Methods =
John W.A. Findlay and Ira Das . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1566
Immunoassay = Stephen G. Schulman and G.J.P.J. Beernink . . . . . . . . . . . . . . . . . . . . . . . . 2048
Radiochemical Methods of Analysis = R. Raghavan and Jose C. Joseph . . . . . . . . . . . . . . . . . 3082
Radiolabeling of Pharmaceutical Aerosols and Gamma Scintigraphic Imaging for
Lung Deposition = Hak-Kim Chan, Stefan Eberl, and William Glover . . . . . . . . . . . . . . 3094

xxxi
 xxxii

Analytical Methods (cont’d. )

Spectroscopic Methods
Flame Photometry = Thomas M. Nowak . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1759
Solid-State NMR in the Characterization of Pharmaceutical Formulations =
Eric J. Munson and Joseph W. Lubach . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3297
Spectroscopic Methods of Analysis: Atomic Absorption and Emission Spectrophotometry =
John P. Oberdier . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3367
Spectroscopic Methods of Analysis: Diffuse Reflectance Spectroscopy =
Herbert Michael Heise . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3375
Spectroscopic Methods of Analysis: Fluorescence Spectroscopy =
Stephen G. Schulman and Jeffrey A. Hughes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3387
Spectroscopic Methods of Analysis: Infrared Spectroscopy = Marilyn D. Duerst . . . . . . . . . . . . 3405
Spectroscopic Methods of Analysis: Mass Spectrometry = Mike S. Lee . . . . . . . . . . . . . . . . . . 3419
Spectroscopic Methods of Analysis: Near-Infrared Spectrometry = Emil W. Ciurczak . . . . . . . . . 3434
Spectroscopic Methods of Analysis: Nuclear Magnetic Resonance Spectroscopy =
Thomas M. O’Connell and Kevin L. Facchine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3440
Spectroscopic Methods of Analysis: Ultraviolet and Visible Spectrophotometry =
R. Raghavan and Jose C. Joseph . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3460
Tablet Evaluation Using Near-Infrared Spectroscopy = Christopher T. Rhodes and
Karen Morisseau . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3630

Thermal Methods
Calorimetry in Pharmaceutical Research and Development = Sophie-Dorothe´e Clas,
Chad R. Dalton, and Bruno C. Hancock . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 393
Thermal Analysis of Drugs and Drug Products = Danie`lle Giron . . . . . . . . . . . . . . . . . . . . . . 3726

Bioavailability and Bioequivalence

Absorption Enhancers = J. P. Remon . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Absorption of Drugs = Peter G. Welling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
Bioavailability of Drugs and Bioequivalence = James T. Dalton and Charles R. Yates . . . . . . . . 164
Biopharmaceutics = Leon Shargel and Andrew B.C. Yu . . . . . . . . . . . . . . . . . . . . . . . . . . . 208
Biotransformation of Drugs = Les F. Chasseaud and D. R. Hawkins . . . . . . . . . . . . . . . . . . . 310
Generic Drugs and Generic Equivalency = Arthur H. Kibbe . . . . . . . . . . . . . . . . . . . . . . . . . 1891
Salt Forms: Pharmaceutical Aspects = Owen I. Corrigan . . . . . . . . . . . . . . . . . . . . . . . . . . . 3177

Biotechnology, Biological Products, and Biological Applications

Biologic Fluids: Analysis = Stephen G. Schulman, Judith A. Davis, and
Gayle A. Brazeau . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 194
Biosynthesis of Drugs = Geoffrey A. Cordell and Kyung Hee Lee . . . . . . . . . . . . . . . . . . . . . 228
Biotechnology and Biological Preparations = Ronald P. Evens . . . . . . . . . . . . . . . . . . . . . . . 258
Biotechnology-Derived Drug Products: Formulation Development = Mary E.M. Cromwell . . . . . . 281
Biotechnology-Derived Drug Products: Stability Testing, Filling, and Packaging =
Mary E.M. Cromwell . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 302
Blood Substitutes: Fluorocarbon Approach = Jean G. Riess . . . . . . . . . . . . . . . . . . . . . . . . . 335
 xxxiii

Blood Substitutes: Hemoglobin-Based Oxygen Carriers = Deanna J. Nelson . . . . . . . . . . . . . . . 353

Colloids and Colloid Drug Delivery System = Diane J. Burgess . . . . . . . . . . . . . . . . . . . . . . . 636
DNA Probes for the Identification of Microbes = Wayne P. Olson . . . . . . . . . . . . . . . . . . . . . 929
Drug Delivery: Monoclonal Antibodies = John B. Cannon, Ho-Wah Hui, and
Pramod K. Gupta . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1132
Drug Delivery: Monoclonal Antibodies in Imaging and Therapy = Ban-An Khaw . . . . . . . . . . . . 1149
Enzyme Immunoassay and Related Bioanalytical Methods = John W.A. Findlay and
Ira Das . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1566
Fractal Geometry in Pharmaceutical and Biological Applications = P. Tang,
Hak-Kim Chan, and Judy A. Raper . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1791
Genetic Aspects of Drug Development = Werner Kalow . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1897
Immunoassay = Stephen G. Schulman and G.J.P.J. Beernink . . . . . . . . . . . . . . . . . . . . . . . . 2048
Lead Optimization in Pharmaceutical Development: Molecular and Cellular Approaches =
C. K. Atterwill and M. G. Wing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2192
Peptides and Proteins: Buccal Absorption = Ashim K. Mitra, Hemant H. Alur, and
Thomas P. Johnston . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2664
Peptides and Proteins: Nasal Absorption = Takahiro Morita and Hiroshi Yamahara . . . . . . . . . 2678
Peptides and Proteins: Non-Invasive Delivery = Michael R. DeFelippis,
Muppalla Sukumar, and Natarajan Rajagopalan . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2692
Peptides and Proteins: Oral Absorption = Eugene J. McNally and Jung Y. Park . . . . . . . . . . . . 2713
Peptides and Proteins: Pulmonary Absorption = Igor Gonda . . . . . . . . . . . . . . . . . . . . . . . . . 2731
Peptides and Proteins: Transdermal Absorption = Richard H. Guy,
M. Begon~a Delgado-Charro, and Diego Marro . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2741
Pharmacogenomics and Genomic Technologies = Daniel A. Brazeau . . . . . . . . . . . . . . . . . . . 2794
Plants as Drugs = Christine K. O’Neil and Charles W. Fetrow . . . . . . . . . . . . . . . . . . . . . . . 2901
Proteomics: Pharmaceutical Applications = A. Ian Smith . . . . . . . . . . . . . . . . . . . . . . . . . . . 3041
Vaccines and Other Immunological Products = Suresh K. Mittal, Harm HogenEsch, and
Kinam Park . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3908

Chemical Properties Associated with Pharmaceutical Systems

Autoxidation and Antioxidants = John M. Pezzuto and Eun Jung Park . . . . . . . . . . . . . . . . . . 139
Biosynthesis of Drugs = Geoffrey A. Cordell and Kyung Hee Lee . . . . . . . . . . . . . . . . . . . . . 228
Biotechnology and Biological Preparations = Ronald P. Evens . . . . . . . . . . . . . . . . . . . . . . . 258
Corrosion of Pharmaceutical Equipment = Paul K. Whitcraft . . . . . . . . . . . . . . . . . . . . . . . . 782
Dendrimers = Antony D’Emanuele, David Attwood, and Ragheb Abu-Rmaileh . . . . . . . . . . . . 872
Drug Design: Basic Principles and Applications = Jacques H. Poupaert . . . . . . . . . . . . . . . . . . 1362
Drug Master Files = C. Jeanne Taborsky and Brian James Reamer . . . . . . . . . . . . . . . . . . . 1401
Gelatin-Containing Formulations: Changes in Dissolution Characteristics =
Saranjit Singh and Sariputta P. Pakhale . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1861
Hydrolysis of Drugs = Jason M. LePree and Kenneth A. Connors . . . . . . . . . . . . . . . . . . . . . 2040
Isomerism = Thomas N. Riley, Jack DeRuiter, William R. Ravis, and C. Randall Clark . . . . . . 2142
Lead Optimization in Pharmaceutical Development: Molecular and Cellular Approaches =
C. K. Atterwill and M. G. Wing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2192
Photodecomposition of Drugs = Hanne Hjorth Tonnesen . . . . . . . . . . . . . . . . . . . . . . . . . . . 2859
Process Chemistry in the Pharmaceutical Industry = Kumar G. Gadamasetti and
Ambarish K. Singh . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2993
Salt Forms: Pharmaceutical Aspects = Owen I. Corrigan . . . . . . . . . . . . . . . . . . . . . . . . . . . 3177
 xxxiv

Clinical Aspects of Drug Development and Use

Adverse Drug Reactions = Therese I. Poirier and Robert L. Maher, Jr. . . . . . . . . . . . . . . . . . . 46
Advertising and Promotion of Prescription and Over-the-Counter Drug Products =
Wayne L. Pines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
Alternative Medicines = Kristi L. Lenz . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66
Clinical Data Management Systems = Samuel V. Givens, Debra Barnes,
Victoria Imber, and Barbara Perry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 551
Clinical Evaluation of Drugs = Lynda Sutton, Allen Cato, and Allen Cato III . . . . . . . . . . . . . 560
Clinical Pharmacokinetics and Pharmacodynamics = Leslie Z. Benet and
Laviero Mancinelli . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 572
Clinical Supplies Manufacture: GMP Considerations = David L. Chesney and
Mark L. Balboni . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 591
Dosage Regimens and Dose-Response = Chyung S. Cook and Aziz Karim . . . . . . . . . . . . . . . . 1012
Drug Abuse = Richard B. Seymour and David E. Smith . . . . . . . . . . . . . . . . . . . . . . . . . . . 1038
Drug Delivery: Monoclonal Antibodies in Imaging and Therapy = Ban-An Khaw . . . . . . . . . . . . 1149
Drug Delivery: Tumor-Targeted Systems = Yu Li and Chao-Pin Lee . . . . . . . . . . . . . . . . . . . 1326
Good Clinical Practices (GCPs): An Overview = Richard A. Guarino . . . . . . . . . . . . . . . . . . . 1925
Medication Errors = Diane D. Cousins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2243
Orphan Drugs = Carolyn H. Asbury . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2468
Vaccines and Other Immunological Products = Suresh K. Mittal,
Harm HogenEsch, and Kinam Park . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3908

Computer Use in Pharmaceutical Technology

Computer Systems Validation = Orlando Lopez . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 707
Computer-Assisted Drug Design = J. Phillip Bowen and Michael Cory . . . . . . . . . . . . . . . . . . 714
Computers in Pharmaceutical Technology = Onkaram Basavapathruni . . . . . . . . . . . . . . . . . . 732
Neural Computing and Formulation Optimization = Elizabeth Colbourn and
Raymond C. Rowe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2399
Paperless Documentation Systems = Ellen M. Williams and Michael McKenna . . . . . . . . . . . . 2551
Virtual Screening = Tı´mea Polgár and György M. Keserü . . . . . . . . . . . . . . . . . . . . . . . . . . 4013

Dispersed Systems
Emulsions and Microemulsions = Gillian M. Eccleston . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1548
Gels and Jellies = Clyde M. Ofner III and Cathy M. Klech-Gelotte . . . . . . . . . . . . . . . . . . . 1875
Surfactants in Pharmaceutical Products and Systems = Owen I. Corrigan and
Anne Marie Healy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3583
Suspensions = Robert A. Nash . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3597
Zeta Potential = Luk Chiu Li and Youqin Tian . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4117

Drug Delivery
Absorption of Drugs = Peter G. Welling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
Amorphous Pharmaceutical Systems = Bruno C. Hancock . . . . . . . . . . . . . . . . . . . . . . . . . . 83
Bioavailability of Drugs and Bioequivalence = James T. Dalton and Charles R. Yates . . . . . . . . 164
 xxxv

Biodegradable Polymers as Drug Carriers = Peter Markland and Victor C. Yang . . . . . . . . . . . 176
Biopharmaceutics = Leon Shargel and Andrew B.C. Yu . . . . . . . . . . . . . . . . . . . . . . . . . . . 208
Dendrimers = Antony D’Emanuele, David Attwood, and Ragheb Abu-Rmaileh . . . . . . . . . . . . 872
Dosage Regimens and Dose-Response = Chyung S. Cook and Aziz Karim . . . . . . . . . . . . . . . . 1012
Drug Delivery Systems: Neutron Scattering Studies = M. Jayne Lawrence and
David J. Barlow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1049
Drug Delivery: Buccal Route = James C. McElnay and Carmel M. Hughes . . . . . . . . . . . . . . . 1071
Drug Delivery: Controlled Release = Yie W. Chien and Senshang Lin . . . . . . . . . . . . . . . . . . . 1082
Drug Delivery: Fast-Dissolve Systems = Vikas Agarwal, Bhavesh H. Kothari,
Derek V. Moe, and Rajendra K. Khankari . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1104
Drug Delivery: Liquid Crystals in = Christel C. Mueller-Goymann . . . . . . . . . . . . . . . . . . . . 1115
Drug Delivery: Monoclonal Antibodies = John B. Cannon, Ho-Wah Hui, and
Pramod K. Gupta . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1132
Drug Delivery: Monoclonal Antibodies in Imaging and Therapy = Ban-An Khaw . . . . . . . . . . . . 1149
Drug Delivery: Mucoadhesive Hydrogels = Hans E. Junginger, J. Coos Verhoef, and
Maya Thanou . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1169
Drug Delivery: Nanoparticles = Elias Fattal and Christine Vauthier . . . . . . . . . . . . . . . . . . . 1183
Drug Delivery: Nasal Route = Rene´ Bommer . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . 1201
Drug Delivery: Needle-Free Systems = Toby King . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1209
Drug Delivery: Ophthalmic Route = Masood Chowhan, Alan L. Weiner, and
Haresh Bhagat . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1220
Drug Delivery: Oral Colon-Specific = Vincent H.L. Lee and Suman K. Mukherjee . . . . . . . . . . 1228
Drug Delivery: Oral Route = Brahma N. Singh and Kwon H. Kim . . . . . . . . . . . . . . . . . . . . 1242
Drug Delivery: Parenteral Route = Michael J. Akers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1266
Drug Delivery: Pulmonary Delivery = Michael T. Newhouse . . . . . . . . . . . . . . . . . . . . . . . . . 1279
Drug Delivery: Pulsatile Systems = Till Bussemer and Roland A. Bodmeier . . . . . . . . . . . . . . . 1287
Drug Delivery: Rectal Route = J. Howard Rytting and Joseph A. Fix . . . . . . . . . . . . . . . . . . . 1298
Drug Delivery: Topical and Transdermal Routes = Kenneth A. Walters . . . . . . . . . . . . . . . . . . 1311
Drug Delivery: Tumor-Targeted Systems = Yu Li and Chao-Pin Lee . . . . . . . . . . . . . . . . . . . 1326
Drug Delivery: Vaginal Route = Yie W. Chien and Chi H. Lee . . . . . . . . . . . . . . . . . . . . . . . 1339
Polymeric Delivery Systems for Poorly Soluble Drugs = Kang Moo Huh,
Sang Cheon Lee, Tooru Ooya, and Kinam Park . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2913

Dosage Forms
Capsules, Hard = Brian E. Jones . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 406
Capsules, Soft = David H. Bergstrom, Stephen Tindal, and Wenbin Dang . . . . . . . . . . . . . . . 419
Colloids and Colloid Drug Delivery System = Diane J. Burgess . . . . . . . . . . . . . . . . . . . . . . . 636
Coprecipitates and Melts = Madhu K. Vadnere . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 774
Dosage Form Design: A Physicochemical Approach = Michael B. Maurin and
Anwar A. Hussain . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 939
Dosage Forms and Basic Preparations: History = Robert A. Buerki and
Gregory J. Higby . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 948
Dosage Forms: Lipid Excipients = Alan L. Weiner . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 975
Dosage Forms: Non-Parenterals = Shailesh K. Singh and Venkatesh Naini . . . . . . . . . . . . . . . 988
Dosage Forms: Parenterals = Gayle A. Brazeau, Adam Persky, and Jintana Napaporn . . . . . . . 1001
Drug Delivery: Needle-Free Systems = Toby King . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1209
Drug Delivery: Pulsatile Systems = Till Bussemer and Roland A. Bodmeier . . . . . . . . . . . . . . . 1287
 xxxvi

Drug Delivery (cont’d.)

Dosage Forms (cont’d.)

Drug Delivery: Tumor-Targeted Systems = Yu Li and Chao-Pin Lee . . . . . . . . . . . . . . . . . . . 1326
Effervescent Pharmaceuticals = Nils-Olof Lindberg and Henri Hansson . . . . . . . . . . . . . . . . . 1454
Emulsions and Microemulsions = Gillian M. Eccleston . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1548
Gastro-Retentive Systems = Amnon Hoffman and Bashir A. Qadri . . . . . . . . . . . . . . . . . . . 1850
Geriatric Dosing and Dosage Forms = Cheryl A. Wiens and Carol A. Borynec . . . . . . . . . . . . . 1905
Hydrogels = Marı´a Dolores Blanco, Rosa Maria Olmo, and José Marı´a Teijón . . . . . . . . . . . . 2021
Liquid Oral Preparations = Jagdish Parasrampuria and Stephen William Pitt . . . . . . . . . . . . . 2216
Lozenges = Robert W. Mendes and Hridaya Bhargava . . . . . . . . . . . . . . . . . . . . . . . . . . . 2231
Metered Dose Inhalers = Sandy J.M. Munro and Alan L. Cripps . . . . . . . . . . . . . . . . . . . . . 2269
Pediatric Dosing and Dosage Forms = Rosalie Sagraves . . . . . . . . . . . . . . . . . . . . . . . . . . . 2629
Powders as Dosage Forms = Jean-Marc Aiache and Erick Beyssac . . . . . . . . . . . . . . . . . . . . 2971
Suspensions = Robert A. Nash . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3597
Ultrasonic Nebulizers = Kevin M.G. Taylor and Orla McCallion . . . . . . . . . . . . . . . . . . . . . 3854
Veterinary Dosage Forms = J. Desmond Baggot . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3941

Dosage Forms, Miscellaneous

Alternative Medicines = Kristi L. Lenz . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66
Cosmetics and Their Relation to Drugs = Martin M. Rieger . . . . . . . . . . . . . . . . . . . . . . . . . 798
Non-Prescription Drugs = Sujit S. Sansgiry, Lynn Simpson, Gauri Shringarpure, and
Amit Kulkarni . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2413
Nutraceutical Supplements = G. Brian Lockwood . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2431

Drug Absorption, Distribution, Metabolism, and Elimination

Bioavailability of Drugs and Bioequivalence = James T. Dalton and Charles R. Yates . . . . . . . . 164
Biotransformation of Drugs = Les F. Chasseaud and D. R. Hawkins . . . . . . . . . . . . . . . . . . . 310
Clinical Pharmacokinetics and Pharmacodynamics = Leslie Z. Benet and
Laviero Mancinelli . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 572
Drug Interactions = Daniel A. Hussar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1392
Metabolite Identification in Drug Discovery = Kathleen A. Cox,
Nigel Clarke, and Diane Rindgen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2262
Prodrug Design = Bernard Testa and Joachim Mayer . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3008

Routes of Administration and Physiological Factors

Drug Delivery: Buccal Route = James C. McElnay and Carmel M. Hughes . . . . . . . . . . . . . . . 1071
Drug Delivery: Mucoadhesive Hydrogels = Hans E. Junginger, J. Coos Verhoef, and
Maya Thanou . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1169
Drug Delivery: Nasal Route = Rene´ Bommer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1201
Drug Delivery: Ophthalmic Route = Masood Chowhan, Alan L. Weiner, and
Haresh Bhagat . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1220
Drug Delivery: Oral Colon-Specific = Vincent H.L. Lee and
Suman K. Mukherjee . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1228
Drug Delivery: Oral Route = Brahma N. Singh and Kwon H. Kim . . . . . . . . . . . . . . . . . . . . 1242
 xxxvii

Drug Delivery: Parenteral Route = Michael J. Akers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1266

Drug Delivery: Pulmonary Delivery = Michael T. Newhouse . . . . . . . . . . . . . . . . . . . . . . . . . 1279
Drug Delivery: Rectal Route = J. Howard Rytting and Joseph A. Fix . . . . . . . . . . . . . . . . . . . 1298
Drug Delivery: Topical and Transdermal Routes = Kenneth A. Walters . . . . . . . . . . . . . . . . . . 1311
Drug Delivery: Vaginal Route = Yie W. Chien and Chi H. Lee . . . . . . . . . . . . . . . . . . . . . . . 1339
Otic Preparations = William H. Slattery III . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2475
Peptides and Proteins: Buccal Absorption = Ashim K. Mitra,
Hemant H. Alur, and Thomas P. Johnston . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2664
Peptides and Proteins: Nasal Absorption = Takahiro Morita and Hiroshi Yamahara . . . . . . . . . 2678
Peptides and Proteins: Non-Invasive Delivery = Michael R. DeFelippis,
Muppalla Sukumar, and Natarajan Rajagopalan . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2692
Peptides and Proteins: Oral Absorption = Eugene J. McNally and Jung Y. Park . . . . . . . . . . . . 2713
Peptides and Proteins: Pulmonary Absorption = Igor Gonda . . . . . . . . . . . . . . . . . . . . . . . . . 2731
Peptides and Proteins: Transdermal Absorption = Richard H. Guy,
M. Begon ~a Delgado-Charro, and Diego Marro . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2741
Physiological Factors Affecting Oral Drug Delivery = Clive G. Wilson,
Bridget O’Mahony, and Blythe Lindsay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2866
Transdermal Delivery: Anatomical Site Influence = Nora Y.K. Chew,
Nina F. Wilkins, and Barrie C. Finnin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3814
Transdermal Delivery: Technologies = S. Kevin Li . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3843
Transdermal Delivery: Sonophoresis = Samir S. Mitragotri, Hua Tang,
E. Daniel Blankschtein, and Robert Langer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3828

Drug Development
Animals in Drug Development = Farrel L. Fort . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114
Calorimetry in Pharmaceutical Research and Development = Sophie-Dorothe´e Clas,
Chad R. Dalton, and Bruno C. Hancock . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 393
Clinical Evaluation of Drugs = Lynda Sutton, Allen Cato, and Allen Cato III . . . . . . . . . . . . . 560
Drug Development Management = James E. Tingstad . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1370
Expert Systems in Pharmaceutical Product Development = Raymond C. Rowe and
Ronald J. Roberts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1663
Genetic Aspects of Drug Development = Werner Kalow . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1897
In Vitro–In Vivo Correlation = J.-M. Cardot and Erick Beyssac . . . . . . . . . . . . . . . . . . . . . . 2062
Lead Optimization in Pharmaceutical Development: Molecular and Cellular Approaches =
C. K. Atterwill and M. G. Wing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2192
Outsourcing = Duane B. Lakings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2486
Patents: International Perspective = Stuart R. Suter and Peter J. Giddings . . . . . . . . . . . . . . . 2604
Patents: United States Perspective = Lorie Ann Morgan and Jeffrey Tidwell . . . . . . . . . . . . . 2616
Pharmacokinetic/Pharmacodynamic Modeling and Simulations in Drug Development =
Dawei Xuan and Sally Y. Choe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2802
Pharmacokinetics: Effects of Food and Fasting = Rajesh Krishna and
Bradford K. Jensen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2816
Prodrug Design = Bernard Testa and Joachim Mayer . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3008
RNAi in Drug Development = Dmitry Samarsky, Shelley Hough, Adam Harris,
Eugene Carstea, and Peter Welch . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3147
Veterinary Pharmaceuticals: Factors Influencing Their Development and Use =
Marilyn N. Martinez, Laura Hungerford, and Mark G. Papich . . . . . . . . . . . . . . . . . . . . 3978
 xxxviii

Drug Discovery
Computer-Assisted Drug Design = J. Phillip Bowen and Michael Cory . . . . . . . . . . . . . . . . . . 714
Drug Design: Basic Principles and Applications = Jacques H. Poupaert . . . . . . . . . . . . . . . . . . 1362
Economic Characteristics of the R&D Intensive Pharmaceutical Industry =
Douglas L. Cocks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1450
Metabolite Identification in Drug Discovery = Kathleen A. Cox, Nigel Clarke, and
Diane Rindgen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2262
Patents: International Perspective = Stuart R. Suter and Peter J. Giddings . . . . . . . . . . . . . . . 2604
Patents: United States Perspective = Lorie Ann Morgan and Jeffrey Tidwell . . . . . . . . . . . . . 2616
Receptors for Drugs: Discovery in the Post-Genomic Era = Jeffrey M. Herz and
William J. Thomsen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3108
Virtual Screening = Tı´mea Polgár and György M. Keserü . . . . . . . . . . . . . . . . . . . . . . . . . . 4013

Drug Dissolution and In Vivo/In Vitro Correlations

Dissolution and Dissolution Testing = A. Mark Dyas and Utpal U. Shah . . . . . . . . . . . . . . . . . 908
Gelatin-Containing Formulations: Changes in Dissolution Characteristics =
Saranjit Singh and Sariputta P. Pakhale . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1861
In Vitro–In Vivo Correlation = J.-M. Cardot and Erick Beyssac . . . . . . . . . . . . . . . . . . . . . . 2062

Equipment Used in Pharmaceutical Development and Manufacturing

Continuous Processing of Pharmaceuticals = J. P. Remon and C. Vervaet . . . . . . . . . . . . . . . . 743
Corrosion of Pharmaceutical Equipment = Paul K. Whitcraft . . . . . . . . . . . . . . . . . . . . . . . . 782
Electrical Power Systems for Pharmaceutical Equipment = Joseph Maida . . . . . . . . . . . . . . . . 1482
Equipment Cleaning = Destin A. LeBlanc . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1580
Extrusion and Extruders = J. M. Newton . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1712
Fluid Bed Processes for Forming Functional Particles = Yoshinobu Fukumori and
Hideki Ichikawa . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1773
Homogenization and Homogenizers = Shailesh K. Singh and Venkatesh Naini . . . . . . . . . . . . . 1996
Isolators for Pharmaceutical Application = Gordon J. Farquharson . . . . . . . . . . . . . . . . . . . . 2133
Laminar Airflow Equipment: Applications and Operation = Gregory F. Peters . . . . . . . . . . . . . . 2171
Materials of Construction for Pharmaceutical Equipment = Michelle M. Gonzalez . . . . . . . . . . . 2237
Roller Compaction Technology for the Pharmaceutical Industry =
Ronald W. Miller and Paul J. Sheskey . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3159
Tablet Compression: Machine Theory, Design, and Process Troubleshooting =
Michael J. Bogda . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3611
Tablet Press Instrumentation = Michael Levin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3684
Tooling for Tableting = Annette Bauer-Brandl . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3782

Excipient Use and Testing in Drug Development

Dosage Forms: Lipid Excipients = Alan L. Weiner . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 975
Excipients for Pharmaceutical Dosage Forms = Patrick J. Crowley and Luigi G. Martini . . . . . . 1609
Excipients: Parenteral Dosage Forms and Their Role = Sandeep Nema,
Ron J. Brendel, and Richard Washkuhn . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1622
 xxxix

Excipients: Powders and Solid Dosage Forms = Hak-Kim Chan and Nora Y.K. Chew . . . . . . . . 1646
Excipients: Safety Testing = Marshall Steinberg and Florence K. Kinoshita . . . . . . . . . . . . . . 1656
Pharmaceutical Excipient Testing: Regulatory and Preclinical Perspective = Paul Baldrick . . . . . 2771
Solubilizing Excipients in Pharmaceutical Formulations = Robert G. Strickley . . . . . . . . . . . . . . 3334
Starches and Starch Derivatives = Ann W. Newman, Ronald L. Mueller,
Imre M. Vitez, and Chris C. Kiesnowski . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3476
Super Disintegrants: Characterization and Function = Larry L. Augsburger,
Albert W. Brzeczko, Umang Shah, and Huijeong Ashley Hahm . . . . . . . . . . . . . . . . . . . . 3553
Waxes = Roland A. Bodmeier . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4066

Formulation Aspects of Pharmaceutical Technology

General Formulation Aspects

Biopharmaceutics = Leon Shargel and Andrew B.C. Yu . . . . . . . . . . . . . . . . . . . . . . . . . . . 208
Biotechnology-Derived Drug Products: Formulation Development = Mary E.M. Cromwell . . . . . . 281
Coloring Agents for Use in Pharmaceuticals = David R. Schoneker . . . . . . . . . . . . . . . . . . . . 648
Dental Products = Sebastian G. Ciancio . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 891
Dosage Form Design: A Physicochemical Approach = Michael B. Maurin and
Anwar A. Hussain . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 939
Excipients for Pharmaceutical Dosage Forms = Patrick J. Crowley and Luigi G. Martini . . . . . . 1609
Expert Systems in Pharmaceutical Product Development = Raymond C. Rowe and
Ronald J. Roberts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1663
Flavors and Flavor Modifiers = Thomas L. Reiland and John M. Lipari . . . . . . . . . . . . . . . . . 1763
Fractal Geometry in Pharmaceutical and Biological Applications = P. Tang,
Hak-Kim Chan, and Judy A. Raper . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1791
Neural Computing and Formulation Optimization = Elizabeth Colbourn and
Raymond C. Rowe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2399
Optimization Methods = Gareth A. Lewis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2452
Packaging Materials: Glass = Claudia C. Okeke and Desmond G. Hunt . . . . . . . . . . . . . . . . . 2508
Salt Forms: Pharmaceutical Aspects = Owen I. Corrigan . . . . . . . . . . . . . . . . . . . . . . . . . . . 3177
Solid-State NMR in the Characterization of Pharmaceutical Formulations =
Eric J. Munson and Joseph W. Lubach . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3297
Supercritical Fluid Technology in Pharmaceutical Research = Aaron S. Mayo and
Uday B. Kompella . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3568
Surfactants in Pharmaceutical Products and Systems = Owen I. Corrigan and
Anne Marie Healy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3583
Tonicity = Jaymin C. Shah . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3768
Veterinary Pharmaceuticals: Factors Influencing Their Development and Use =
Marilyn N. Martinez, Laura Hungerford, and Mark G. Papich . . . . . . . . . . . . . . . . . . . . 3978
Waxes = Roland A. Bodmeier . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4066

Formulation Aspects Relevant to Liquids and Semi-Solids

Buffers, Buffering Agents, and Ionic Equilibria = Harry G. Brittain . . . . . . . . . . . . . . . . . . . . 385
Coacervation and Phase Separation = Bruno Gander, Maria Jose Blanco-Prı´eto,
C. Thomasin, Ch. Wandrey, and D. Hunkeler . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 600
Complexation: Cyclodextrins = Gerold Mosher and Diane O. Thompson . . . . . . . . . . . . . . . . 671
Complexation: Non-Cyclodextrins = Galina N. Kalinkova . . . . . . . . . . . . . . . . . . . . . . . . . . 697
 xl

Formulation Aspects of Pharmaceutical Technology (cont’d.)

Formulation Aspects Relevant to Liquids and Semi-Solids (cont’d.)

Cosolvents and Cosolvency = Joseph T. Rubino . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 806
Dosage Forms: Lipid Excipients = Alan L. Weiner . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 975
Drug Delivery: Liquid Crystals in = Christel C. Mueller-Goymann . . . . . . . . . . . . . . . . . . . . 1115
Excipients: Parenteral Dosage Forms and Their Role = Sandeep Nema,
Ron J. Brendel, and Richard Washkuhn . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1622
Gels and Jellies = Clyde M. Ofner III and Cathy M. Klech-Gelotte . . . . . . . . . . . . . . . . . . . 1875
Inhalation: Liquids = Michael E. Placke, Jeffrey Ding, and William C. Zimlich, Jr. . . . . . . . . . 2092
Iontophoresis = J. Bradley Phipps, Erik R. Scott, J. Richard Gyory, and
Rama V. Padmanabhan . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2119
Rheology of Pharmaceutical Systems = Fridrun Podczeck . . . . . . . . . . . . . . . . . . . . . . . . . . 3128
Solubilization of Drugs in Aqueous Media = Paul B. Myrdal and
Samuel H. Yalkowsky . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3311
Solubilizing Excipients in Pharmaceutical Formulations = Robert G. Strickley . . . . . . . . . . . . . . 3334

Formulation Aspects Relevant to Particles and Solids

Adsorption at Solid Surfaces: Pharmaceutical Applications = Hong Wen . . . . . . . . . . . . . . . . . 34
Amorphous Pharmaceutical Systems = Bruno C. Hancock . . . . . . . . . . . . . . . . . . . . . . .. . . 83
Cocrystals: Design, Properties and Formation Mechanisms = Naı´r Rodrı´guez-Hornedo,
Sarah J. Nehm, and Adivaraha Jayasankar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 615
Cooling Processes and Congealing = Richard Turton and Xiu Xiu Cheng . . . . . . . . . . . . . . . . 761
Coprecipitates and Melts = Madhu K. Vadnere . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 774
Crystal Habit Changes and Dosage Form Performance = A. K. Tiwary . . . . . . . . . . . . . . . . . . 820
Crystallization: General Principles and Significance on Product Development =
Naı´r Rodrı´guez-Hornedo, Ron C. Kelly, Brent D. Sinclair, and Jonathan M. Miller . . . . . . . 834
Dissolution and Dissolution Testing = A. Mark Dyas and Utpal U. Shah . . . . . . . . . . . . . . . . . 908
Drug Delivery: Controlled Release = Yie W. Chien and Senshang Lin . . . . . . . . . . . . . . . . . . . 1082
Drug Delivery: Fast-Dissolve Systems = Vikas Agarwal, Bhavesh H. Kothari,
Derek V. Moe, and Rajendra K. Khankari . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1104
Dry Powder Aerosols: Emerging Technologies = Hak-Kim Chan and Nora Y.K. Chew . . . . . . . . 1428
Excipients: Powders and Solid Dosage Forms = Hak-Kim Chan and Nora Y.K. Chew . . . . . . . . 1646
Film Coating of Oral Solid Dosage Forms = Linda A. Felton . . . . . . . . . . . . . . . . . . . . . . . . 1729
Gelatin-Containing Formulations: Changes in Dissolution Characteristics =
Saranjit Singh and Sariputta P. Pakhale . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1861
Hot-Melt Extrusion Technology = Jim W. McGinity, Michael A. Repka,
John J. Koleng, Jr., and Feng Zhang . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2004
Inhalation: Dry Powder = Lynn Van Campen and Geraldine Venthoye . . . . . . . . . . . . . . . . . 2077
Melt Processes for Oral Solid Dosage Forms = Paul W.S. Heng and Tin Wui Wong . . . . . . . . . . 2257
Microencapsulation Technology = Kinam Park and Yoon Yeo . . . . . . . . . . . . . . . . . . . . . . . . 2315
Microsphere Technology and Applications = Diane J. Burgess and Anthony J. Hickey . . . . . . . . 2328
Polymorphism: Pharmaceutical Aspects = Harry G. Brittain . . . . . . . . . . . . . . . . . . . . . . . . . 2935
Solids: Flow Properties = Stephen A. Howard . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3275
Starches and Starch Derivatives = Ann W. Newman, Ronald L. Mueller,
Imre M. Vitez, and Chris C. Kiesnowski . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3476
 xli

Super Disintegrants: Characterization and Function = Larry L. Augsburger,

Albert W. Brzeczko, Umang Shah, and Huijeong Ashley Hahm . . . . . . . . . . . . . . . . . . . . 3553
Tablet Formulation = Larry L. Augsburger and Mark J. Zellhofer . . . . . . . . . . . . . . . . . . . . 3641
Tablet Testing = Saeed A. Qureshi . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3707
X-Ray Powder Diffractometry = Raj Suryanarayanan and Suneel Rastogi . . . . . . . . . . . . . . . 4103
Zeta Potential = Luk Chiu Li and Youqin Tian . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4117

Genomics and Drug Discovery and Development

Genetic Aspects of Drug Development = Werner Kalow . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1897
Pharmacogenomics and Genomic Technologies = Daniel A. Brazeau . . . . . . . . . . . . . . . . . . . 2794
Proteomics: Pharmaceutical Applications = A. Ian Smith . . . . . . . . . . . . . . . . . . . . . . . . . . . 3041
Receptors for Drugs: Discovery in the Post-Genomic Era = Jeffrey M. Herz and
William J. Thomsen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3108
RNAi in Drug Development = Dmitry Samarsky, Shelley Hough, Adam Harris,
Eugene Carstea, and Peter Welch . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3147

Good Clinical, Laboratory, and Manufacturing Practices

Clinical Supplies Manufacture: GMP Considerations = David L. Chesney and
Mark L. Balboni . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 591
Food and Drug Administration: Role in Drug Regulation = Roger L. Williams . . . . . . . . . . . . . 1779
Good Clinical Practices (GCPs): An Overview = Richard A. Guarino . . . . . . . . . . . . . . . . . . . 1925
Good Laboratory Practices (GLPs): An Overview = Nigel J. Dent . . . . . . . . . . . . . . . . . . . . . 1931
Good Manufacturing Practices (GMPs): An Overview = Ira R. Berry . . . . . . . . . . . . . . . . . . . 1941

Management of Pharmaceutical Systems and Procedures

Clinical Data Management Systems = Samuel V. Givens, Debra Barnes,
Victoria Imber, and Barbara Perry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 551
Drug Development Management = James E. Tingstad . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1370
Expert Systems in Pharmaceutical Product Development = Raymond C. Rowe and
Ronald J. Roberts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1663
Laboratory Information Management System (LIMS) = Siri H. Segalstad . . . . . . . . . . . . . . . . 2164
Outsourcing = Duane B. Lakings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2486
Paperless Documentation Systems = Ellen M. Williams and Michael McKenna . . . . . . . . . . . . 2551
Project Management = Jerome J. Groen and Cara R. Frosch . . . . . . . . . . . . . . . . . . . . . . . . 3015
Quality Systems Management = Gary C. Harbour and Robert G. Kieffer . . . . . . . . . . . . . . . . 3075

Manufacture of Pharmaceutical Products

Capsules, Hard = Brian E. Jones . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 406
Capsules, Soft = David H. Bergstrom, Stephen Tindal, and Wenbin Dang . . . . . . . . . . . . . . . 419
Clinical Supplies Manufacture: GMP Considerations = David L. Chesney and
Mark L. Balboni . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 591
Continuous Processing of Pharmaceuticals = J. P. Remon and C. Vervaet . . . . . . . . . . . . . . . . 743
 xlii

Manufacture of Pharmaceutical Products (cont’d.)

Contract Manufacturing = Duncan E. McVean . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 750
Drug Master Files = C. Jeanne Taborsky and Brian James Reamer . . . . . . . . . . . . . . . . . . . 1401
Drying and Dryers = Anthony J. Hlinak and Bradley A. Clark . . . . . . . . . . . . . . . . . . . . . . 1435
Elastomeric Components for the Pharmaceutical Industry = Edward J. Smith . . . . . . . . . . . . . . 1466
Electrical Power Systems for Pharmaceutical Equipment = Joseph Maida . . . . . . . . . . . . . . . . 1482
Evaporation and Evaporators = David P. Kessler . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1600
Extrusion and Extruders = J. M. Newton . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1712
Film Coating of Oral Solid Dosage Forms = Linda A. Felton . . . . . . . . . . . . . . . . . . . . . . . . 1729
Fluid Bed Processes for Forming Functional Particles = Yoshinobu Fukumori and
Hideki Ichikawa . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1773
Freeze Drying, Scale-Up Considerations = Edward H. Trappler . . . . . . . . . . . . . . . . . . . . . . 1834
Good Manufacturing Practices (GMPs): An Overview = Ira R. Berry . . . . . . . . . . . . . . . . . . . 1941
Handling Hazardous Chemicals and Pharmaceuticals = Antonio Conto . . . . . . . . . . . . . . . . . . 1948
Homogenization and Homogenizers = Shailesh K. Singh and Venkatesh Naini . . . . . . . . . . . . . 1996
Mixing and Segregation in Tumbling Blenders = Troy Shinbrot and Fernando J. Muzzio . . . . . . . 2352
Packaging Materials: Glass = Claudia C. Okeke and Desmond G. Hunt . . . . . . . . . . . . . . . . . 2508
Pilot Plant Design = Mickey L. Wells, Samuel B. Balik, Ralph B. Caricofe,
Charles W. Crew, Ronald A. Sanftleben, and A. Wayne Wood . . . . . . . . . . . . . . . . . . . . 2875
Pilot Plant Operation = Mickey L. Wells, A. Wayne Wood, Samuel B. Balik,
Ralph B. Caricofe, Ronald A. Sanftleben, and Charles W. Crew . . . . . . . . . . . . . . . . . . . 2886
Process Chemistry in the Pharmaceutical Industry = Kumar G. Gadamasetti and
Ambarish K. Singh . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 2993
Roller Compaction Technology for the Pharmaceutical Industry =
Ronald W. Miller and Paul J. Sheskey . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3159
Scale-Up and Post Approval Changes (SUPAC) = Jerome P. Skelly . . . . . . . . . . . . . . . . . . . . 3188
Scale-Up of Solid Dosage Forms = Colleen E. Ruegger, Alan Royce, Matthew J. Mollan, Jr.,
Robert Wagner, Stephen Valazza, and Mark Mecadon . . . . . . . . . . . . . . . . . . . . . . . . . . 3193
Tablet Manufacture = Norman Anthony Armstrong . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3653
Tablet Manufacture by Direct Compression = Norman Anthony Armstrong . . . . . . . . . . . . . . . 3673
Tablet Press Instrumentation = Michael Levin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3684
Technology Transfer Considerations for Pharmaceuticals = Ira R. Berry . . . . . . . . . . . . . . . . . 3717
Unit Processes in Pharmacy: Fundamentals = Anthony J. Hickey and
David Ganderton . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3862
Unit Processes in Pharmacy: Operations = Anthony J. Hickey and David Ganderton . . . . . . . . 3879

Oral Delivery of Drugs

Drug Delivery: Buccal Route = James C. McElnay and Carmel M. Hughes . . . . . . . . . . . . . . . 1071
Drug Delivery: Oral Colon-Specific = Vincent H.L. Lee and Suman K. Mukherjee . . . . . . . . . . 1228
Drug Delivery: Oral Route = Brahma N. Singh and Kwon H. Kim . . . . . . . . . . . . . . . . . . . . 1242
Film Coating of Oral Solid Dosage Forms = Linda A. Felton . . . . . . . . . . . . . . . . . . . . . . . . 1729
Gastro-Retentive Systems = Amnon Hoffman and Bashir A. Qadri . . . . . . . . . . . . . . . . . . . 1850
Liquid Oral Preparations = Jagdish Parasrampuria and Stephen William Pitt . . . . . . . . . . . . . 2216
Lozenges = Robert W. Mendes and Hridaya Bhargava . . . . . . . . . . . . . . . . . . . . . . . . . . . 2231
 xliii

Peptides and Proteins: Oral Absorption = Eugene J. McNally and Jung Y. Park . . . . . . . . . . . . 2713
Physiological Factors Affecting Oral Drug Delivery = Clive G. Wilson,
Bridget O’Mahony, and Blythe Lindsay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2866

Organizations Involved in Pharmaceuticals

European Agency for the Evaluation of Medicinal Products (EMEA) = David M. Jacobs . . . . . . 1593
Food and Drug Administration: Role in Drug Regulation = Roger L. Williams . . . . . . . . . . . . . 1779
Harmonization of Pharmacopeial Standards = Lee T. Grady and Jerome A. Halperin . . . . . . . . 1955
Health Care Systems: Outside the United States = Albert I. Wertheimer and
Sheldon X. Kong . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1977
Health Care Systems: Within the United States = Henri R. Manasse, Jr. . . . . . . . . . . . . . . . . . 1985
Pharmacopeial Standards: European Pharmacopeia = Agnès Artiges . . . . . . . . . . . . . . . . . . . . 2829
Pharmacopeial Standards: Japanese Pharmacopeia = Mitsuru Uchiyama . . . . . . . . . . . . . . . . . 2836
Pharmacopeial Standards: United States Pharmacopeia and National Formulary =
Lee T. Grady . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2841
World Health Organization (WHO): Global Harmonization of Requirements for Medicinal
Products = Juhana E. Idanpaan-Heikkila . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4099

Packaging of Pharmaceuticals
Blow-Fill-Seal: Advanced Aseptic Processing = Deborah J. Jones . . . . . . . . . . . . . . . . . . . . . . 378
Elastomeric Components for the Pharmaceutical Industry = Edward J. Smith . . . . . . . . . . . . . . 1466
Extractables and Leachables in Drugs and Packaging = Daniel L. Norwood,
Alice T. Granger, and Diane M. Paskiet . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1693
Packaging Materials: Glass = Claudia C. Okeke and Desmond G. Hunt . . . . . . . . . . . . . . . . . 2508
Packaging Systems: Compendial Requirements = Claudia C. Okeke, Desmond G. Hunt,
Nicholas Mohr, Thomas Medwick, Eric B. Sheinin, and Roger L. Williams . . . . . . . . . . . . 2526
Water Sorption of Drugs and Dosage Forms = Mark J. Kontny and James J. Conners . . . . . . . . 4049

Parenteral Delivery of Drugs

Dosage Forms: Parenterals = Gayle A. Brazeau, Adam Persky, and Jintana Napaporn . . . . . . . 1001
Drug Delivery: Needle-Free Systems = Toby King . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1209
Drug Delivery: Parenteral Route = Michael J. Akers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1266
Pyrogens and Endotoxin Detection = James F. Cooper . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3052
Sterilization: Dry Heat = Andrea Chieppo and Thomas Kupiec . . . . . . . . . . . . . . . . . . . . . . 3512
Sterilization: Ethylene Oxide = Terezinha de Jesus Andreo Pinto . . . . . . . . . . . . . . . . . . . . . 3519
Sterilization: Moist Heat = Dario Pistolesi . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3529
Sterilization: Radiation = Stephen G. Schulman and Phillip M. Achey . . . . . . . . . . . . . . . . . . 3540
Viral Inactivation Issues in Aseptically Processed Parenterals = J. Hotta, A. Klos,
S. Petteway, Jr., and D. Pifat . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3997

Particle Size Characterization, Modification and Significance

Colloids and Colloid Drug Delivery System = Diane J. Burgess . . . . . . . . . . . . . . . . . . . . . . . 636
Crystal Habit Changes and Dosage Form Performance = A. K. Tiwary . . . . . . . . . . . . . . . . . . 820
 xliv

Particle Size Characterization, Modification and Significance (cont’d.)

Crystallization: General Principles and Significance on Product Development =
Naı´r Rodrı´guez-Hornedo, Ron C. Kelly, Brent D. Sinclair, and Jonathan M. Miller . . . . . . . 834
Crystallization: Particle Size Control = Richard D. Braatz, Mitsuko Fujiwara,
Thomas J. Wubben, and Effendi Rusli . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 858
Drug Delivery: Nanoparticles = Elias Fattal and Christine Vauthier . . . . . . . . . . . . . . . . . . . 1183
Dry Powder Aerosols: Emerging Technologies = Hak-Kim Chan and Nora Y.K. Chew . . . . . . . . 1428
Microsphere Technology and Applications = Diane J. Burgess and Anthony J. Hickey . . . . . . . . 2328
Milling of Active Pharmaceutical Ingredients = Elizabeth S. Fisher . . . . . . . . . . . . . . . . . . . . 2339
Nanoparticle Engineering = Robert O. Williams III and Jason M. Vaughn . . . . . . . . . . . . . . . . 2384
Particle Engineering = Miriam K. Franchini . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2567
Particle-Size Characterization = Brian H. Kaye . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2582
Pelletization Techniques = Isaac Ghebre-Sellassie and Axel Knoch . . . . . . . . . . . . . . . . . . . . 2651
Secondary Electron Microscopy in Pharmaceutical Technology = Peter C. Schmidt . . . . . . . . . . 3217
Supercritical Fluid Technology in Pharmaceutical Research = Aaron S. Mayo and
Uday B. Kompella . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3568
X-Ray Powder Diffractometry = Raj Suryanarayanan and Suneel Rastogi . . . . . . . . . . . . . . . 4103

Peptides and Proteins in Pharmaceuticals

Genetic Aspects of Drug Development = Werner Kalow . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1897
Peptides and Proteins: Buccal Absorption = Ashim K. Mitra,
Hemant H. Alur, and Thomas P. Johnston . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2664
Peptides and Proteins: Nasal Absorption = Takahiro Morita and
Hiroshi Yamahara . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2678
Peptides and Proteins: Non-Invasive Delivery = Michael R. DeFelippis,
Muppalla Sukumar, and Natarajan Rajagopalan . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2692
Peptides and Proteins: Oral Absorption = Eugene J. McNally and Jung Y. Park . . . . . . . . . . . . 2713
Peptides and Proteins: Pulmonary Absorption = Igor Gonda . . . . . . . . . . . . . . . . . . . . . . . . . 2731
Peptides and Proteins: Transdermal Absorption = Richard H. Guy,
M. Begon ~a Delgado-Charro, and Diego Marro . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2741
Pharmacogenomics and Genomic Technologies = Daniel A. Brazeau . . . . . . . . . . . . . . . . . . . 2794
Protein Binding of Drugs = Sylvie Laganière and Iain J. McGilveray . . . . . . . . . . . . . . . . . . 3027
Proteomics: Pharmaceutical Applications = A. Ian Smith . . . . . . . . . . . . . . . . . . . . . . . . . . . 3041
Receptors for Drugs: Discovery in the Post-Genomic Era = Jeffrey M. Herz and
William J. Thomsen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3108
RNAi in Drug Development = Dmitry Samarsky, Shelley Hough, Adam Harris,
Eugene Carstea, and Peter Welch . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3147

Pharmacokinetics and Pharmacodynamics

Biotransformation of Drugs = Les F. Chasseaud and D. R. Hawkins . . . . . . . . . . . . . . . . . . . 310
Clinical Pharmacokinetics and Pharmacodynamics = Leslie Z. Benet and
Laviero Mancinelli . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 572
Geriatric Dosing and Dosage Forms = Cheryl A. Wiens and Carol A. Borynec . . . . . . . . . . . . . 1905
Pediatric Dosing and Dosage Forms = Rosalie Sagraves . . . . . . . . . . . . . . . . . . . . . . . . . . . 2629
 xlv

Pharmacokinetic/Pharmacodynamic Modeling and Simulations in Drug Development =

Dawei Xuan and Sally Y. Choe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2802
Pharmacokinetics: Effects of Food and Fasting = Rajesh Krishna and Bradford K. Jensen . . . . . 2816
Population Pharmacokinetics = Ene I. Ette, Alaa M. Ahmad, and Paul J. Williams . . . . . . . . . 2946
Protein Binding of Drugs = Sylvie Laganière and Iain J. McGilveray . . . . . . . . . . . . . . . . . . 3027

Physical Properties Associated with Pharmaceutical Systems

Buffers, Buffering Agents, and Ionic Equilibria = Harry G. Brittain . . . . . . . . . . . . . . . . . . . . 385
Coacervation and Phase Separation = Bruno Gander, Maria Jose Blanco-Prı´eto,
C. Thomasin, Ch. Wandrey, and D. Hunkeler . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 600
Cocrystals: Design, Properties and Formation Mechanisms = Naı´r Rodrı´guez-Hornedo,
Sarah J. Nehm, and Adivaraha Jayasankar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 615
Complexation: Cyclodextrins = Gerold Mosher and Diane O. Thompson . . . . . . . . . . . . . . . . 671
Complexation: Non-Cyclodextrins = Galina N. Kalinkova . . . . . . . . . . . . . . . . . . . . . . . . . . 697
Cooling Processes and Congealing = Richard Turton and Xiu Xiu Cheng . . . . . . . . . . . . . . . . 761
Coprecipitates and Melts = Madhu K. Vadnere . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 774
Cosolvents and Cosolvency = Joseph T. Rubino . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 806
Crystal Habit Changes and Dosage Form Performance = A. K. Tiwary . . . . . . . . . . . . . . . . . . 820
Crystallization: General Principles and Significance on Product Development =
Naı´r Rodrı´guez-Hornedo, Ron C. Kelly, Brent D. Sinclair, and Jonathan M. Miller . . . . . . . 834
Crystallization: Particle Size Control = Richard D. Braatz, Mitsuko Fujiwara,
Thomas J. Wubben, and Effendi Rusli . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 858
Dissolution and Dissolution Testing = A. Mark Dyas and Utpal U. Shah . . . . . . . . . . . . . . . . . 908
Dosage Form Design: A Physicochemical Approach = Michael B. Maurin and
Anwar A. Hussain . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 939
Drug Delivery: Controlled Release = Yie W. Chien and Senshang Lin . . . . . . . . . . . . . . . . . . . 1082
Drug Delivery: Fast-Dissolve Systems = Vikas Agarwal, Bhavesh H. Kothari,
Derek V. Moe, and Rajendra K. Khankari . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1104
Drug Delivery: Liquid Crystals in = Christel C. Mueller-Goymann . . . . . . . . . . . . . . . . . . . . 1115
Effervescent Pharmaceuticals = Nils-Olof Lindberg and Henri Hansson . . . . . . . . . . . . . . . . . 1454
Electrostatic Charge in Pharmaceutical Systems = Philip Chi Lip Kwok and
Hak-Kim Chan . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1535
Fractal Geometry in Pharmaceutical and Biological Applications = P. Tang,
Hak-Kim Chan, and Judy A. Raper . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1791
Freeze Drying = Michael J. Pikal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1807
Gastro-Retentive Systems = Amnon Hoffman and Bashir A. Qadri . . . . . . . . . . . . . . . . . . . 1850
Particle-Size Characterization = Brian H. Kaye . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2582
Partition Coefficients = Eric J. Lien and Steven Shijun Ren . . . . . . . . . . . . . . . . . . . . . . . . 2595
Polymorphism: Pharmaceutical Aspects = Harry G. Brittain . . . . . . . . . . . . . . . . . . . . . . . . . 2935
Rheology of Pharmaceutical Systems = Fridrun Podczeck . . . . . . . . . . . . . . . . . . . . . . . . . . 3128
Solids: Flow Properties = Stephen A. Howard . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3275
Supercritical Fluid Technology in Pharmaceutical Research = Aaron S. Mayo and
Uday B. Kompella . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3568
Tonicity = Jaymin C. Shah . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3768
X-Ray Powder Diffractometry = Raj Suryanarayanan and Suneel Rastogi . . . . . . . . . . . . . . . 4103
Zeta Potential = Luk Chiu Li and Youqin Tian . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4117
 xlvi

Polymers used in Pharmaceutical Systems and Technology

Bioabsorbable Polymers = Shalaby W. Shalaby . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155
Biodegradable Polymers as Drug Carriers = Peter Markland and Victor C. Yang . . . . . . . . . . . 176
Dendrimers = Antony D’Emanuele, David Attwood, and Ragheb Abu-Rmaileh . . . . . . . . . . . . 872
Drug Delivery: Mucoadhesive Hydrogels = Hans E. Junginger, J. Coos Verhoef, and
Maya Thanou . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1169
Elastomeric Components for the Pharmaceutical Industry = Edward J. Smith . . . . . . . . . . . . . . 1466
Gels and Jellies = Clyde M. Ofner III and Cathy M. Klech-Gelotte . . . . . . . . . . . . . . . . . . . 1875
Hydrogels = Marı´a Dolores Blanco, Rosa Maria Olmo, and José Marı´a Teijón . . . . . . . . . . . . 2021
Polymeric Delivery Systems for Poorly Soluble Drugs = Kang Moo Huh,
Sang Cheon Lee, Tooru Ooya, and Kinam Park . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2913
Polymers in Transdermal Delivery Systems = Fumio Kamiyama and Ying-shu Quan . . . . . . . . . 2925

Pulmonary Delivery of Pharmaceuticals

Drug Delivery: Pulmonary Delivery = Michael T. Newhouse . . . . . . . . . . . . . . . . . . . . . . . . . 1279
Dry Powder Aerosols: Emerging Technologies = Hak-Kim Chan and Nora Y. K. Chew . . . . . . . . 1428
Inhalation: Dry Powder = Lynn Van Campen and Geraldine Venthoye . . . . . . . . . . . . . . . . . 2077
Inhalation: Liquids = Michael E. Placke, Jeffrey Ding, and William C. Zimlich, Jr. . . . . . . . . . 2092
Metered Dose Inhalers = Sandy J.M. Munro and Alan L. Cripps . . . . . . . . . . . . . . . . . . . . . 2269
Peptides and Proteins: Pulmonary Absorption = Igor Gonda . . . . . . . . . . . . . . . . . . . . . . . . . 2731
Radiolabeling of Pharmaceutical Aerosols and Gamma Scintigraphic Imaging for
Lung Deposition = Hak-Kim Chan, Stefan Eberl, and William Glover . . . . . . . . . . . . . . 3094
Ultrasonic Nebulizers = Kevin M.G. Taylor and Orla McCallion . . . . . . . . . . . . . . . . . . . . . 3854

Quality Assurance and Control

Pharmaceutical Quality Assurance Microbiology Laboratories = Anthony M. Cundell . . . . . . . . 2783
Quality Assurance of Pharmaceuticals = Barbara Immel . . . . . . . . . . . . . . . . . . . . . . . . . . . 3064
Quality Systems Management = Gary C. Harbour and Robert G. Kieffer . . . . . . . . . . . . . . . . 3075

Regulatory, Legal, and Compendial Aspects and Requirements

21 CFR Part 11 Revisited = Thomas Linz and Sigrun Seeger . . . . . . . . . . . . . . . . . . . . . . . . 1
Adverse Drug Reactions = Therese I. Poirier and Robert L. Maher, Jr. . . . . . . . . . . . . . . . . . . 46
Advertising and Promotion of Prescription and Over-the-Counter Drug Products =
Wayne L. Pines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
Alternative Medicines = Kristi L. Lenz . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66
Carcinogenicity Testing: Past, Present, and Future = Koen Van Deun . . . . . . . . . . . . . . . . . . . 431
Clinical Data Management Systems = Samuel V. Givens, Debra Barnes,
Victoria Imber, and Barbara Perry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 551
Clinical Evaluation of Drugs = Lynda Sutton, Allen Cato, and Allen Cato III . . . . . . . . . . . . . 560
Coloring Agents for Use in Pharmaceuticals = David R. Schoneker . . . . . . . . . . . . . . . . . . . . 648
Contract Manufacturing = Duncan E. McVean . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 750
Cosmetics and Their Relation to Drugs = Martin M. Rieger . . . . . . . . . . . . . . . . . . . . . . . . . 798
Drug Information Systems = Linda Lisgarten and Michelle Wake . . . . . . . . . . . . . . . . . . . . . 1385
 xlvii

Drug Master Files = C. Jeanne Taborsky and Brian James Reamer . . . . . . . . . . . . . . . . . . . 1401
Drug Safety Evaluation = Farrel L. Fort . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1406
Equipment Cleaning = Destin A. LeBlanc . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1580
European Agency for the Evaluation of Medicinal Products (EMEA) = David M. Jacobs . . . . . . 1593
Excipients: Safety Testing = Marshall Steinberg and Florence K. Kinoshita . . . . . . . . . . . . . . 1656
Expiration Dating = Leslie C. Hawley and Mark D. VanArendonk . . . . . . . . . . . . . . . . . . . . 1685
Food and Drug Administration: Role in Drug Regulation = Roger L. Williams . . . . . . . . . . . . . 1779
Generic Drugs and Generic Equivalency = Arthur H. Kibbe . . . . . . . . . . . . . . . . . . . . . . . . . 1891
Harmonization of Pharmacopeial Standards = Lee T. Grady and Jerome A. Halperin . . . . . . . . 1955
Non-Prescription Drugs = Sujit S. Sansgiry, Lynn Simpson, Gauri Shringarpure, and
Amit Kulkarni . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2413
Nutraceutical Supplements = G. Brian Lockwood . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2431
Orphan Drugs = Carolyn H. Asbury . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2468
Packaging Systems: Compendial Requirements = Claudia C. Okeke, Desmond G. Hunt,
Nicholas Mohr, Thomas Medwick, Eric B. Sheinin, and Roger L. Williams . . . . . . . . . . . . 2526
Paperless Documentation Systems = Ellen M. Williams and Michael McKenna . . . . . . . . . . . . 2551
Pharmaceutical Excipient Testing: Regulatory and Preclinical Perspective = Paul Baldrick . . . . . 2771
Pharmacopeial Standards: European Pharmacopeia = Agnès Artiges . . . . . . . . . . . . . . . . . . . . 2829
Pharmacopeial Standards: Japanese Pharmacopeia = Mitsuru Uchiyama . . . . . . . . . . . . . . . . . 2836
Pharmacopeial Standards: United States Pharmacopeia and National Formulary =
Lee T. Grady . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2841
Scale-Up and Post Approval Changes (SUPAC) = Jerome P. Skelly . . . . . . . . . . . . . . . . . . . . 3188

Safety Considerations of Drugs and Pharmaceutical Products

Absorption Enhancers = J. P. Remon . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Adsorption at Solid Surfaces: Pharmaceutical Applications = Hong Wen . . . . . . . . . . . . . . . . . 34
Animals in Drug Development = Farrel L. Fort . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114
Autoxidation and Antioxidants = John M. Pezzuto and Eun Jung Park . . . . . . . . . . . . . . . . . . 139
Carcinogenicity Testing: Past, Present, and Future = Koen Van Deun . . . . . . . . . . . . . . . . . . . 431
Coloring Agents for Use in Pharmaceuticals = David R. Schoneker . . . . . . . . . . . . . . . . . . . . 648
Drug Abuse = Richard B. Seymour and David E. Smith . . . . . . . . . . . . . . . . . . . . . . . . . . . 1038
Drug Design: Basic Principles and Applications = Jacques H. Poupaert . . . . . . . . . . . . . . . . . . 1362
Drug Information Systems = Linda Lisgarten and Michelle Wake . . . . . . . . . . . . . . . . . . . . . 1385
Drug Interactions = Daniel A. Hussar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1392
Drug Safety Evaluation = Farrel L. Fort . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1406
Excipients: Safety Testing = Marshall Steinberg and Florence K. Kinoshita . . . . . . . . . . . . . . 1656
Expiration Dating = Leslie C. Hawley and Mark D. VanArendonk . . . . . . . . . . . . . . . . . . . . 1685
Extractables and Leachables in Drugs and Packaging = Daniel L. Norwood,
Alice T. Granger, and Diane M. Paskiet . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1693
Handling Hazardous Chemicals and Pharmaceuticals = Antonio Conto . . . . . . . . . . . . . . . . . . 1948
Isolators for Pharmaceutical Application = Gordon J. Farquharson . . . . . . . . . . . . . . . . . . . . 2133
Medication Errors = Diane D. Cousins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2243
Microbial Control of Pharmaceuticals = Nigel A. Halls . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2286
Non-Prescription Drugs = Sujit S. Sansgiry, Lynn Simpson, Gauri Shringarpure, and
Amit Kulkarni . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2413
 xlviii

Safety Considerations of Drugs and Pharmaceutical Products (cont’d.)

Pharmaceutical Excipient Testing: Regulatory and Preclinical Perspective =
Paul Baldrick . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2771
Plants as Drugs = Christine K. O’Neil and Charles W. Fetrow . . . . . . . . . . . . . . . . . . . . . . . 2901
Pyrogens and Endotoxin Detection = James F. Cooper . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3052
Trace Level Impurity Analysis = Daniel L. Norwood, Fenghe Qiu, and James O. Mullis . . . . . . . 3797

Scale-Up and Pilot Plant Design and Operation

Freeze Drying, Scale-Up Considerations = Edward H. Trappler . . . . . . . . . . . . . . . . . . . . . . 1834
Pilot Plant Design = Mickey L. Wells, Samuel B. Balik, Ralph B. Caricofe,
Charles W. Crew, Ronald A. Sanftleben, and A. Wayne Wood . . . . . . . . . . . . . . . . . . . . 2875
Pilot Plant Operation = Mickey L. Wells, A. Wayne Wood, Samuel B. Balik,
Ralph B. Caricofe, Ronald A. Sanftleben, and Charles W. Crew . . . . . . . . . . . . . . . . . . . 2886
Scale-Up and Post Approval Changes (SUPAC) = Jerome P. Skelly . . . . . . . . . . . . . . . . . . . . 3188
Scale-Up of Solid Dosage Forms = Colleen E. Ruegger, Alan Royce, Matthew J. Mollan, Jr.,
Robert Wagner, Stephen Valazza, and Mark Mecadon . . . . . . . . . . . . . . . . . . . . . . . . . . 3193
Technology Transfer Considerations for Pharmaceuticals = Ira R. Berry . . . . . . . . . . . . . . . . . 3717
Wet Granulation: End-Point Determination and Scale-Up = Michael Levin . . . . . . . . . . . . . . . 4078

Semi-Solids as Pharmaceuticals
Capsules, Soft = David H. Bergstrom, Stephen Tindal, and Wenbin Dang . . . . . . . . . . . . . . . 419
Rheology of Pharmaceutical Systems = Fridrun Podczeck . . . . . . . . . . . . . . . . . . . . . . . . . . 3128
Semisolid Preparations = Guru Betageri and Sunil Prabhu . . . . . . . . . . . . . . . . . . . . . . . . . 3257
Waxes = Roland A. Bodmeier . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4066

Solids and Powders as Pharmaceuticals

Adsorption at Solid Surfaces: Pharmaceutical Applications = Hong Wen . . . . . . . . . . . . . . . . . 34
Amorphous Pharmaceutical Systems = Bruno C. Hancock . . . . . . . . . . . . . . . . . . . . . . . . . . 83
Capsules, Hard = Brian E. Jones . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 406
Cocrystals: Design, Properties and Formation Mechanisms = Naı´r Rodrı´guez-Hornedo,
Sarah J. Nehm, and Adivaraha Jayasankar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 615
Crystal Habit Changes and Dosage Form Performance = A. K. Tiwary . . . . . . . . . . . . . . . . . . 820
Effervescent Pharmaceuticals = Nils-Olof Lindberg and Henri Hansson . . . . . . . . . . . . . . . . . 1454
Powder Sampling = Helena J. Venables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2959
Powders as Dosage Forms = Jean-Marc Aiache and Erick Beyssac . . . . . . . . . . . . . . . . . . . . 2971
Scale-Up of Solid Dosage Forms = Colleen E. Ruegger, Alan Royce, Matthew J. Mollan, Jr.,
Robert Wagner, Stephen Valazza, and Mark Mecadon . . . . . . . . . . . . . . . . . . . . . . . . . . 3193
Solids: Flow Properties = Stephen A. Howard . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3275
Tablet Formulation = Larry L. Augsburger and Mark J. Zellhofer . . . . . . . . . . . . . . . . . . . . 3641
Tablet Manufacture = Norman Anthony Armstrong . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3653
 xlix

Solubilization and Solubilized Systems

Cosolvents and Cosolvency = Joseph T. Rubino . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 806
Polymeric Delivery Systems for Poorly Soluble Drugs = Kang Moo Huh, Sang Cheon Lee,
Tooru Ooya, and Kinam Park . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2913
Solubilization of Drugs in Aqueous Media = Paul B. Myrdal and Samuel H. Yalkowsky . . . . . . . 3311
Solubilizing Excipients in Pharmaceutical Formulations = Robert G. Strickley . . . . . . . . . . . . . . 3334
Surfactants in Pharmaceutical Products and Systems = Owen I. Corrigan and
Anne Marie Healy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3583

Stability and Preservation of Drugs and Drug Products

Autoxidation and Antioxidants = John M. Pezzuto and Eun Jung Park . . . . . . . . . . . . . . . . . . 139
Biotechnology-Derived Drug Products: Stability Testing, Filling, and Packaging =
Mary E.M. Cromwell . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 302
Expiration Dating = Leslie C. Hawley and Mark D. VanArendonk . . . . . . . . . . . . . . . . . . . . 1685
Headspace Oxygen Analysis in Pharmaceutical Products = Allen C. Templeton and
Robert A. Reed . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1967
Hydrolysis of Drugs = Jason M. LePree and Kenneth A. Connors . . . . . . . . . . . . . . . . . . . . . 2040
Microbial Control of Pharmaceuticals = Nigel A. Halls . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2286
Moisture in Pharmaceutical Products = R. Gary Hollenbeck . . . . . . . . . . . . . . . . . . . . . . . . 2368
Photodecomposition of Drugs = Hanne Hjorth Tonnesen . . . . . . . . . . . . . . . . . . . . . . . . . . . 2859
Preservation of Pharmaceutical Products = Peter Gilbert and David G. Allison . . . . . . . . . . . . 2983
Water Sorption of Drugs and Dosage Forms = Mark J. Kontny and James J. Conners . . . . . . . . 4049

Statistics
Statistical Methods = Charles Bon . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3483
Statistical Process Control and Process Capability = Thomas D. Murphy,
Shailesh K. Singh, and Merlin L. Utter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3499

Sterile Products and Microbiological Monitoring and Control

Aseptic Processing: Validation = James P. Agalloco and James E. Akers . . . . . . . . . . . . . . . . 127
Bio-Validation of Steam Sterilization = Nigel A. Halls . . . . . . . . . . . . . . . . . . . . . . . . . . . . 325
Blow-Fill-Seal: Advanced Aseptic Processing = Deborah J. Jones . . . . . . . . . . . . . . . . . . . . . . 378
DNA Probes for the Identification of Microbes = Wayne P. Olson . . . . . . . . . . . . . . . . . . . . . 929
Dressings in Wound Management = Sarah M.E. Cockbill . . . . . . . . . . . . . . . . . . . . . . . . . . 1023
Filters and Filtration = Maik W. Jornitz . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1748
Isolators for Pharmaceutical Application = Gordon J. Farquharson . . . . . . . . . . . . . . . . . . . . 2133
Laminar Airflow Equipment: Applications and Operation = Gregory F. Peters . . . . . . . . . . . . . . 2171
Microbiologic Monitoring of Controlled Processes = Gregory F. Peters and
Marghi R. McKeon . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2298
Pharmaceutical Quality Assurance Microbiology Laboratories = Anthony M. Cundell . . . . . . . . 2783
Pyrogens and Endotoxin Detection = James F. Cooper . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3052
Sterilization: Dry Heat = Andrea Chieppo and Thomas Kupiec . . . . . . . . . . . . . . . . . . . . . . 3512
Sterilization: Ethylene Oxide = Terezinha de Jesus Andreo Pinto . . . . . . . . . . . . . . . . . . . . . 3519
 l

Sterile Products and Microbiological Monitoring and Control (cont’d.)

Sterilization: Moist Heat = Dario Pistolesi . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3529
Sterilization: Radiation = Stephen G. Schulman and Phillip M. Achey . . . . . . . . . . . . . . . . . . 3540
Viral Inactivation Issues in Aseptically Processed Parenterals = J. Hotta, A. Klos,
S. Petteway, Jr., and D. Pifat . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3997

Tableting of Pharmaceuticals
Starches and Starch Derivatives = Ann W. Newman, Ronald L. Mueller,
Imre M. Vitez, and Chris C. Kiesnowski . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3476
Super Disintegrants: Characterization and Function = Larry L. Augsburger, Albert W. Brzeczko,
Umang Shah, and Huijeong Ashley Hahm . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3553
Tablet Compression: Machine Theory, Design, and Process Troubleshooting =
Michael J. Bogda . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3611
Tablet Evaluation Using Near-Infrared Spectroscopy = Christopher T. Rhodes and
Karen Morisseau . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3630
Tablet Formulation = Larry L. Augsburger and Mark J. Zellhofer . . . . . . . . . . . . . . . . . . . . 3641
Tablet Manufacture = Norman Anthony Armstrong . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3653
Tablet Manufacture by Direct Compression = Norman Anthony Armstrong . . . . . . . . . . . . . . . 3673
Tablet Testing = Saeed A. Qureshi . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3707
Tooling for Tableting = Annette Bauer-Brandl . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3782
Wet Granulation: End-Point Determination and Scale-Up = Michael Levin . . . . . . . . . . . . . . . 4078

Topical and Transdermal Delivery and Application

Cosmetics and Their Relation to Drugs = Martin M. Rieger . . . . . . . . . . . . . . . . . . . . . . . . . 798
Dressings in Wound Management = Sarah M.E. Cockbill . . . . . . . . . . . . . . . . . . . . . . . . . . 1023
Drug Delivery: Topical and Transdermal Routes = Kenneth A. Walters . . . . . . . . . . . . . . . . . . 1311
Iontophoresis = J. Bradley Phipps, Erik R. Scott, J. Richard Gyory, and
Rama V. Padmanabhan . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2119
Peptides and Proteins: Transdermal Absorption = Richard H. Guy,
M. Begon ~a Delgado-Charro, and Diego Marro . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2741
Polymers in Transdermal Delivery Systems = Fumio Kamiyama and Ying-shu Quan . . . . . . . . . 2925
Transdermal Delivery: Anatomical Site Influence = Nora Y.K. Chew, Nina F. Wilkins, and
Barrie C. Finnin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3814
Transdermal Delivery: Technologies = S. Kevin Li . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3843
Transdermal Delivery: Sonophoresis = Samir S. Mitragotri, Hua Tang,
E. Daniel Blankschtein, and Robert Langer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3828

Unit Operations and Processing Techniques Employed in Pharmaceutical Development and

Manufacturing
Continuous Processing of Pharmaceuticals = J. P. Remon and C. Vervaet . . . . . . . . . . . . . . . . 743
Cooling Processes and Congealing = Richard Turton and Xiu Xiu Cheng . . . . . . . . . . . . . . . . 761
Drying and Dryers = Anthony J. Hlinak and Bradley A. Clark . . . . . . . . . . . . . . . . . . . . . . 1435
Evaporation and Evaporators = David P. Kessler . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1600
Extrusion and Extruders = J. M. Newton . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1712
 li

Filters and Filtration = Maik W. Jornitz . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1748

Fluid Bed Processes for Forming Functional Particles = Yoshinobu Fukumori and
Hideki Ichikawa . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1773
Freeze Drying = Michael J. Pikal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1807
Freeze Drying, Scale-Up Considerations = Edward H. Trappler . . . . . . . . . . . . . . . . . . . . . . 1834
Homogenization and Homogenizers = Shailesh K. Singh and Venkatesh Naini . . . . . . . . . . . . . 1996
Hot-Melt Extrusion Technology = Jim W. McGinity, Michael A. Repka,
John J. Koleng, Jr., and Feng Zhang . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2004
Melt Processes for Oral Solid Dosage Forms = Paul W.S. Heng and Tin Wui Wong . . . . . . . . . . 2257
Microencapsulation Technology = Kinam Park and Yoon Yeo . . . . . . . . . . . . . . . . . . . . . . . . 2315
Microsphere Technology and Applications = Diane J. Burgess and Anthony J. Hickey . . . . . . . . 2328
Milling of Active Pharmaceutical Ingredients = Elizabeth S. Fisher . . . . . . . . . . . . . . . . . . . . 2339
Mixing and Segregation in Tumbling Blenders = Troy Shinbrot and Fernando J. Muzzio . . . . . . . 2352
Nanoparticle Engineering = Robert O. Williams III and Jason M. Vaughn . . . . . . . . . . . . . . . . 2384
Optimization Methods = Gareth A. Lewis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2452
Particle Engineering = Miriam K. Franchini . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2567
Pelletization Techniques = Isaac Ghebre-Sellassie and Axel Knoch . . . . . . . . . . . . . . . . . . . . 2651
Roller Compaction Technology for the Pharmaceutical Industry =
Ronald W. Miller and Paul J. Sheskey . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3159
Unit Processes in Pharmacy: Fundamentals = Anthony J. Hickey and David Ganderton . . . . . . . 3862
Unit Processes in Pharmacy: Operations = Anthony J. Hickey and David Ganderton . . . . . . . . 3879

Validation of Pharmaceutical Processes and Techniques

Analytical Procedures: Validation = Joachim Ermer and John S. Landy . . . . . . . . . . . . . . . . . 92
Aseptic Processing: Validation = James P. Agalloco and James E. Akers . . . . . . . . . . . . . . . . 127
Bio-Validation of Steam Sterilization = Nigel A. Halls . . . . . . . . . . . . . . . . . . . . . . . . . . . . 325
Computer Systems Validation = Orlando Lopez . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 707
Equipment Cleaning = Destin A. LeBlanc . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1580
Filters and Filtration = Maik W. Jornitz . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1748
Validation of Pharmaceutical Processes = Robert A. Nash . . . . . . . . . . . . . . . . . . . . . . . . . . 3928

Water in Pharmaceutical Manufacturing and Products

Moisture in Pharmaceutical Products = R. Gary Hollenbeck . . . . . . . . . . . . . . . . . . . . . . . . 2368
Water for Pharmaceuticals = Leonid Shnayder . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4039
Water Sorption of Drugs and Dosage Forms = Mark J. Kontny and James J. Conners . . . . . . . . 4049
Wet Granulation: End-Point Determination and Scale-Up = Michael Levin . . . . . . . . . . . . . . . 4078
Preface

The introductory paragraph of the preface to both the first and second editions of this
encyclopedia, published in 1988 and 2002, respectively, notes that pharmaceutical science
and technology have progressed enormously in recent years, and that significant advances
in therapeutics and an understanding of the need to optimize drug delivery in the body
have brought about an increased awareness of the valuable role played by the dosage form
in therapy. In turn, this has resulted in an increased sophistication and level of expertise in
the design, development, manufacture, testing and regulation of drugs and dosage forms.
This statement is as true today as it was back in 1988 and 2002—and perhaps more so,
given the increasing emphasis being placed on the discovery, development, and use of large
molecular entities as therapeutic and diagnostic agents. The pace at which these advances
are being made is reflected in the fact that, after only four years, it has been felt necessary
to publish this, the third edition, of the Encyclopedia of Pharmaceutical Technology. As
with the second edition, the third edition is available in print and also online.
The third edition continues the focus on the discovery, development, design, manufac-
ture, testing, regulation, and commercialization of drugs and dosage forms. Areas of
emphasis include pharmaceutics, pharmacokinetics, analytical chemistry, quality assur-
ance, drug safety, and manufacturing processes. Both more traditional and newer technol-
ogies and processes are included, with an increased emphasis on biotechnology and large
molecule development. Current trends relating to solid state aspects of drug entities are
also included, reflecting again the advances made in this area.
While of primary interest to pharmaceutical scientists and management in the pharma-
ceutical and related industries, including regulatory agencies, the encyclopedia will be of
value to those in academia undertaking pharmaceutical research and those responsible
for the education and training in pharmaceutical science and technology of graduate
and undergraduate students.
The print version of the third edition consists of six volumes totaling about 4400 pages,
an approximately 45% increase in content compared to the second edition. The number of
articles has increased to almost 300 titles arranged alphabetically by subject and the num-
ber of contributors has risen by over 50% to in excess of 500 individuals. The new edition
now contains a Topical Table of Contents, whose purpose is to group article titles
into categories and subcategories, thereby making the reader aware of other related and
relevant articles.
As was the case with the second edition, the online version includes everything in the
print version and also offers the convenience of a keyword search engine as well as the
inclusion of color illustrations. New and revise articles will be digitally posted quarterly
and available to all subscribers of the electronic version.
Preparation of the third edition began under the auspices of Marcel Dekker, Inc. before
it became part of Informa Healthcare USA, Inc. last year. I would be remiss therefore if I
did not acknowledge the fantastic support received from Carolyn Hall, Managing Editor
of the Encyclopedia Department of Marcel Dekker, Inc. prior to the merger. At the same
time, it is a pleasure to acknowledge to contributions to the development of this third
edition from staff in The Encyclopedia Group of Informa Healthcare USA, Inc., in parti-
cular Louisa Lam, Claire Miller, and Yvonne Honigsberg.
I must note the absence of Jim Boylan’s name as an editor on the third edition. Jim and
I worked closely in partnership as co-editors on both the first and second editions. After
17 years commitment to the encyclopedia Jim decided to relinquish his role as co-editor,

liii
 liv

a move that both I and the publisher greatly regretted. And so, it is fitting that this new
edition, which relies in large part on Jim’s past contributions, is dedicated to him.
Finally, you the readers are to be thanked for your support and comments. I trust you
will find that the third edition continues the high standards set by the previous editions. As
always, I welcome your comments and suggestions for new titles.

James Swarbrick, Editor

Pinehurst, NC
About the Editor
JAMES SWARBRICK is President of PharmaceuTech, Inc., a consulting company located in Pinehurst, North
Carolina. During his more than 40 year career in the area of pharmaceutical science he has had both extensive
academic and industrial working experience. He has also served on a number of regulatory and compendial
committees, as well as serving as a consultant to pharmaceutical companies and organizations in the Americas,
Europe and Australia. From 1993 through early 2006 he was VP for R&D and then VP for Scientific Affairs at
AAIPharma in Wilmington, North Carolina. Prior to joining AAIPharma, he was, for 12 years, Professor and
Chairman of the Division of Pharmaceutics at the University of North Carolina at Chapel Hill School of Pharmacy.
Other academic positions held include Professorships at the University of Connecticut, the University of Sydney in
Australia, the University of Southern California, and Dean of the School of Pharmacy at the University of London.
He also served as Director of Product Development at the Sterling-Winthrop Research Institute, Rensselaer, NY,
a guest scientist at Astra Laboratories in Sweden, and Visiting Professor of Pharmaceutics at Shanghai Medical
University in China. Other appointments include chairman of the Joint USP-NF Panel on Dissolution and
Disintegration Testing, and a four-year term as a member (and then chairman) of the Generic Drugs Advisory
Committee (now Advisory Committee for Pharmaceutical Science) of the Food and Drug Administration (FDA).
He currently serves as chairman of the Pharmaceutics Advisory Committee of the Pharmaceutical Research and
Manufacturers of America (PhRMA) Foundation and is a member of that organization’s Scientific Advisory
Committee. He is a Fellow of the American Association of Pharmaceutical Scientists, the Academy of
Pharmaceutical Sciences, the Royal Society of Chemistry, and the Royal Pharmaceutical Society of Great Britain.
Dr. Swarbrick received the B.Pharm. degree (1960), the Ph.D. degree (1964) in pharmaceutics and the D.Sc.
degree (1972) in surface and physical chemistry from London University, England.

lv
 Encyclopedia of

Volume 6
Third Edition

Statistical–Zeta
Pages 3483 through 4128
Pharmaceutical Technology

Water–Zeta Veterinary– Unit–Validation Trace–Ultrasonic Technology– Tablet–Tablet Super– Statistical–

Virtual Tooling Suspensions Sterilization
 Sterilization
Statistical–
Statistical Methods
Charles Bon
AAI Pharma, Inc., Wilmington, North Carolina, U.S.A.

INTRODUCTION the extract is reconstituted in 100 ml of mobile phase,

an acetonitrile/methanol/buffer solution, and 10 ml is
Death and taxes, as the old adage goes, are the only injected onto a high-pressure liquid chromatogaph
certainties in life. Although this is an overstatement, (HPLC) equipped with a 3 m, C8 column. The column
it does emphasize the uncertain world in which the effluent is monitored for ultraviolet absorbance at
pharmaceutical scientist lives and works. Faced with 280 nm, and the drug and internal standard peak
‘‘estimates’’ of product characteristics such as potency, area, or height, responses are determined. The drug-
content uniformity, impurity levels, and dissolution to-internal peak response ratio is calculated, and an
performance, the scientist must make important go/ estimate of the concentration of drug in each specimen
no-go decisions. These estimated values, as is true for is obtained through interpolation of the ratio on a
most measured quantities, inherently vary from the calibration curve. The calibration curve is constructed
true values on which correct decisions depend. Statistical from the peak response ratios of extracted calibrator
methods provide tools that enable the pharmaceutical specimens containing known amounts of the drug in
scientist to act decisively in an uncertain world. An interference-free blood.
understanding of basic statistical methods is important There are several obvious sources where error can
for all who work in the pharmaceutical field. contribute to the uncertainty in the concentration
estimate. If, for example, 1.0 ml of the specimen is
mistakenly transferred instead of 0.5 ml, but the correct
SOME BASIC CONCEPTS volume was used for preparing the calibrators, system-
atic error would have occurred. Systematic errors lead
In the developmental process for many drugs, the drug to a constant, predictable uncertainty in the estimate.
product is administered under controlled conditions to To deal with a systematic error, we must recognize that
healthy, normal individuals or to the targeted patient it has occurred and then correct for the mistake that led
population. This is done to characterize the rate and to its occurrence. Statistical methods do not address
extent of absorption, the bioavailability, of the active systematic errors. If, however, the correct 0.5 ml-
drug contained in the product. The bioavailability is volume of specimen and 100 ml volume of internal stan-
estimated from the measured concentrations of the dard were used, we still would have errors affecting the
drug that appear in serial blood specimens collected concentration estimate. These errors would be random
over a period of time after product administration. errors. Random errors are positive and negative devia-
Basic statistical principles govern the behavior of the tions that inherently occur in any attempt to exactly
typical bioanalytical procedure used to measure these measure a quantity, in our case, specific volumes of
concentrations in the collected blood specimens. the specimen and internal standard. We might transfer
The first step of a typical procedure might involve 0.498 ml of a specimen owing to an unrecognized air
the transfer of 0.5 ml of the specimen into a screw- bubble interfering with our reading of the meniscus
cap culture tube using a disposable serological pipette. in the transfer pipette. Or, perhaps, 0.502 ml is trans-
Next, 100 ml of internal standard, a chemical structur- ferred because of some of the specimen adhering
ally similar to the drug of interest, is added. A small to the outside of the pipette. Statistical methods are
volume, 1.0 ml, of a buffer solution at an appropriate our tools for dealing with uncertainty resulting from
pH level is added to decrease the aqueous solubility random error (chance).
of the drug and internal standard. After thorough mix- After the transfer of the specimen and the addition
ing, 7 ml of diethyl ether is added to the tube, and it is of internal standard, the drug-to-internal ratio
capped and shaken to extract the drug and internal becomes a fixed quantity. Any additional random or
standard into the organic ether phase. The tube is cen- systematic volume errors should not affect the concen-
trifuged to obtain a clean separation of the aqueous tration estimate. Upon injection of the reconstituted
and the organic phases. A 5 ml portion of the organic extract onto the HPLC, however, random errors will
phase is transferred to a clean culture tube and iseva- occur that do affect the estimate. These are the result
porated to dryness under nitrogen. The residual of of chance deviations in the partitioning of the drug
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-100000437
Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved. 3483
 3484 Statistical Methods
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Statistical–

and internal standard between themobile phase and the desirable property for any statistic. Some statistics only
column. Random fluctuations in the UV detection sys- become unbiased when sample sizes are large. These
tem will also affect the concentration estimate. The statistics are said to be asymptotically unbiased. Such
influences of random errors are statistically additive. statistics have their greatest utility when used with lar-
In our example, the random errors are independent ger samples, such as those consisting of 20–30 determi-
of each other in that the occurrence, sign (positive or nations rather than smaller numbers such as 2–10.
negative), and magnitude of each are unrelated to the Reproducibility, or precision, of a method relates to
occurrence, sign, and magnitude of the others. This is how individual estimates fluctuate around the average
in contrast to correlated errors, which are related to value. The magnitude of the fluctuation in the popu-
each other in some predictable way. lation is expressed by the parameter variance (s2).
Two important characteristics of any assay method Variance is the average of the squared deviations about
are its accuracy and its reproducibility. Accuracy is m for all values xi in the population: S(xi  m)2/N. An
how close an estimate is expected to be in relation to unbiased estimate of s2 is obtained from the deviation
the true value for the specimen. Reproducibility relates of each value (xi) around the mean (x
) for a sample
to how repeated estimates of the same specimen vary taken from the population: s2 ¼ S(xi  x
)2/
about their average value. Both accuracy and repro- (n  1) ¼ (Sxi2  (Sxi)2/n)/(n  1).
ducibility are usually defined for a given concentration The form of the equation not involving x
is a con-
and may differ between low and high specimen concen- venient calculating formula that avoids rounding pro-
trations. In a good assay method, these differences blems that can occur when individual values are very
should be inconsequential across the working range close to themean. It is common practice to report the
of the method. standard deviation s, which is the square root of the
The expected value of an estimate is the average of sample variance. The standard deviation is often nor-
an infinite number of determinations of the estimate. malized as the percent coefficient of variation CV%
These infinite determinations, taken as an aggregate, by dividing it by thesample mean and expressing the
make up the population of estimates. A population result as a percent. In the analyses of pharmaceutical
does not have to be infinite in size. Some examples of dosage units and in FDA regulations governing these
finite populations are the potencies for all tablets in a analyses, the term relative standard deviation (RSD)
given lot of a drug product or the sitting blood pres- is used for this calculated quantity instead of the
sures of all patients who use a certain antihypertensive term CV%.
medication in the coming year. The mean (m) of a Given two estimates of a statistic, one from a sam-
population is a parameter of the population. The ple of size n and the other from a sample of size 2n,
estimate of m, obtained from a single concentration one might expect that the estimate from the larger sam-
estimate for a specimen, varies from one determination ple would be more reliable than that from the smaller
to the next and is aptly referred to as a variable. If a sample. This is, in fact, supported by statistical theory.
variable conceptually takes on a continuum of values, If the variance in the population is s2, then the
as is the case for a concentration estimate, it is called variance of the sample mean for samples of size n is
a continuous variable. Variables that take on only s2/n. The square root of this is the standard error of
certain discrete values, such as the number of tablets the mean. Consistent with the variance of the sample
produced from 50 kg of active drug material, are mean being 1/n times that of a single determination
referred to as discrete variables. (s2), the standard deviation and the CV% of the
It is impossible to conduct an infinite number of sample mean are reduced by the square root of n. As
extractions of a speciman to determine the accuracy a direct consequence, an assay method that relies on
of a method. As a result, we estimate the accuracy of the mean of two independent
p concentration determina-
an assay by performing a finite number of extractions tions has a CV 1/ 2 that of the same method based on
(n) on the specimen. We report the accuracy as the a single determination. This provides an easy way to
mean (x
¼ Sxi/n, i ¼ 1, 2, . . . , n) of the multiple increase the precision (reduce variability) of a method.
determinations, expressed as a percent of the known An example of this is found in radioimmunoassay in
concentration. The finite group of determinations is a which it is common for a concentration estimate to
sample from the population, and its mean is referred be calculated from the mean response of two determi-
to as the sample mean. The sample mean is a statistic nations of a specimen.
that estimates the population parameter m. If we could As noted previously, fluctuations in concentration
obtain the means from an infinite number of same-size estimates about the true value arise from multiple,
samples, regardless of their size, then the mean of these independent, random errors. Each of the independent
infinite sample means would equal m. In statistical ter- errors (s2i ) is statistically additive, such that the total
minology, we say that the sample mean is an unbiased assay error s2 ¼ Ss2i . Similarly, the CV% of the assay
estimator of the population mean. Unbiasedness is a will be the square root of the sum of the squared CV%
Statistical Methods 3485

Sterilization
Statistical–
values for all independent sources of error in the the second, s, defines the spread of the distribution
method. It is interesting to examine the impact of this about its center. The distribution has some unique
on the determination of the important pharmacoki- properties. Its mean is the same as its median, the value
netic measure Cmax, the maximum concentration of a at which 50% of the population are below and the
drug after administration of a drug product. For many remaining 50% are above. The distribution is sym-
drugs, the biological variability, the degree to which metrical, its shape below its center is the mirror image
the true Cmax value varies during replicate administra- of its shape above its center. Its mode, the value that
tions of the product to the same individual, can have a occurs with the greatest frequency, also coincides with
CV of 25% or greater. If an assay method has a 10% the mean and median. Approximately 68% of the
CV and the biological variability for Cmax is 25%, then distribution lies within 1s of the mean, 95% lies within
the Cmax estimate would have a CV of 26.9%. This is 2s of the mean, and 99.8% lies within 3s of the mean.
simply the square root of the sum of the two squared The behavior of many observations in nature and
independent error CVs: 252 þ 102. If the precision of many measurements in science can be approximated
the assay method was improved to 5% CV, Cmax would using the normal distribution. An important property
be estimated with a 25.5% CV. The effort required to that leads to the nearly universal application of the dis-
reduce a 10% CV assay to half that level, to obtain a tribution is found in the central limit theorem. This
mere 1.4% increase in overall precision would seldom theorem states that regardless of whether a given popu-
prove to be cost-effective. lation is normally distributed, the distribution of the
mean of randomly selected samples from the popu-
lation will tend toward normality. This tendency
A USEFUL STATISTICAL DISTRIBUTION increases as the size of the sample increases. If the
population is, in reality, normally distributed, then a
The normal distribution appears ubiquitously through- sample size of 1 is all that is needed. The more deviant
out science and nature. References to applications of the population distribution is from normality, the
normal theory are found throughout the pharmaceuti- larger the sample size needs to be for its mean to be
cal literature. The distribution is one of the earliest normally distributed.
introduced, having been published in 1733 by De It is reasonable to question whether the distribution
Moivre. Most scientists have a basic familiarity with of the estimates of a drug concentration in a blood
the distribution and its characteristic bell-shaped curve specimen might be approximated by the normal distri-
(Fig. 1). Those familiar with column chromatography bution. Table 1 presents the results of repeated analy-
might recognize this shape as that of the perfect chro- ses of a specimen of interference-free plasma spiked to
matographic peak. In fact, the principles of chromato- contain a known amount of drug. These data are taken
graphic peak symmetry and peak-to-peak resolution from a comparative bioavailability study in which sin-
can be derived from normal theory. The normal distri- gle doses of an unmarketed generic product and the
bution is defined by two parameters. The first, m, marketed brand product of a drug were administered
defines the central location of the distribution, and on separate occasions to healthy males. The values pre-
sented are the first of duplicate determinations of a
quality control (QC) specimen that was included with
each batch of subject specimens. This was done to ver-
0.40 ify that the in-process accuracy and precision of the
assay method were consistent with the values observed
0.35 during the assay validation.
A frequency histogram of the results is shown at the
0.30
top of Fig. 2. The bottom of the figure shows the plot
0.25 of the normal distribution with mean 207.6 ng/ml and
Frequency

s 14.1 ng/ml. The shape of the histogram plot is similar

0.20
to the plot of the normal distribution. The greatest
0.15 deviation between the two is in the region of the center
histogram. The sample distribution is higher peaked in
0.10 its center, containing 42% of the values, than is the nor-
mal distribution, which has a 38% frequency at its cen-
0.05
ter. The mean, median, and mode of the QC values are
0.00 close to being equal, as would be expected if the values
–4.00 –3.00 –2.00 –1.00 0.00 1.00 2.00 3.00 4.00
Sigma units from mean
had come from a normal distribution.
A particularly useful form of the normal distri-
Fig. 1 The normal distribution. bution is obtained by transforming each value xi to
 3486 Statistical Methods
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Table 1 Results of quality control specimen analyses 45.0%

Batch Conc. (ng/ml) Batch Conc. (ng/ml) 40.0%
1 188 24 201
35.0%

Percent of Distribution
2 216 25 197
30.0%
3 201 26 199
4 166 27 180 25.0%

5 214 28 237 20.0%

6 209 29 212 15.0%
7 226 30 239
10.0%
8 183 31 207
5.0%
9 210 32 216
10 213 33 213 0.0%
172 154 196 208 220 232 244
11 209 34 226
Value (ng/ml)
12 222 35 213
13 214 36 204 45.0%

14 213 37 194 40.0%

15 205 38 218

Percent of Distribution
35.0%
16 226 39 207 30.0%
17 203 40 196
25.0%
18 188 41 208
20.0%
19 215 42 210
20 211 Mean 207.6 15.0%

21 201 Median 209 10.0%

22 205 Mode 213
5.0%
23 206 Standard deviation 14.1
0.0%
166 172 178 184 190 196 202 208 214 220 226 232 238 244 250
Value (ng/ml)

its standard normal value. This transformation Fig. 2 QC frequency histogram plot and corresponding
converts the distribution to one that is independent normal distribution.
of m and s. The conversion is Zi ¼ (xi  m)/s, where
Zi is known as the standard normal deviate and is
normally distributed with m ¼ 0 and s ¼ 1. Tables calculations. The lower limit and upper limit values,
of Z-values can be found in any elementary statistics in ng/ml, for each range is calculated as: [x
 Z
s],
textbook, An example is presented in Table 2. The [x
þ Z
s].
standard normal deviate table typically provides the As seen in the frequency histogram, the observed
proportion (area under the curve) of the distribution distribution of the QC values in the vicinity of the
that lies between 1 and various Z-values (the lower mean, between Z ¼ 1 and Z ¼ 1, is higher than
tail) or between various Z-values and þ1 (the upper predicted by normal theory. However, the distribution
tail). The proportion of the distribution lying within a outside this region closely resembles what would be
given Z range around m is calculated by taking the differ- expected for a normally distributed variable. A
ence between the tabled proportions for þZ and Z. goodness-of-fit test to determine whether a variable
Table 3 provides the expected percentages of the follows a certain statistical distribution can be con-
standard normal distribution that lie within some structed using a chi-square statistic (Table 4). The
selected Z ranges about m and compares these with range of the sample values is divided into intervals,
the percentage of the QC sample distribution that falls and the expected number of values (E) that should fall
within these ranges. The percentages for the sample in each interval is calculated. It is important to keep
distribution are calculated by taking the number of the intervals large enough so that at least five obser-
QC values within each range, dividing it by 42, the vations are expected in each. The number of observed
total number of values in the sample, and expressing values (O) in the sample that falls within each interval
this as a percent. Because m and s are unknown, the is then determined. The chi-square statistic for this test
sample mean and standard deviation are used in the is w2 ¼ S[(O  E)2/E]. If the calculated statistic value
Statistical Methods 3487

Sterilization
Statistical–
Table 2 Cumulative areas under the standard normal curve (1 to Z)ab
Z 0.00 0.01 0.02 0.03 0.04 0.05 0.06 0.07 0.08 0.09
0.0 0.5000 0.5040 0.5080 0.5120 0.5160 0.5199 0.5239 0.5279 0.5319 0.5359
0.1 0.5398 0.5438 0.5478 0.5517 0.5557 0.5596 0.5636 0.5675 0.5714 0.5753
0.2 0.5793 0.5832 0.5871 0.5910 0.5948 0.5987 0.6026 0.6064 0.6103 0.6141
0.3 0.6179 0.6217 0.6255 0.6293 0.6331 0.6368 0.6406 0.6443 0.6480 0.6517
0.4 0.6554 0.6591 0.6628 0.6664 0.6700 0.6736 0.6772 0.6808 0.6844 0.6879
0.5 0.6915 0.6950 0.6985 0.7019 0.7054 0.7088 0.7123 0.7157 0.7190 0.7224
0.6 0.7257 0.7291 0.7324 0.7357 0.7389 0.7422 0.7454 0.7486 0.7517 0.7549
0.7 0.7580 0.7611 0.7642 0.7673 0.7704 0.7734 0.7764 0.7794 0.7823 0.7852
0.8 0.7881 0.7910 0.7939 0.7967 0.7995 0.8023 0.8051 0.8078 0.8106 0.8133
0.9 0.8159 0.8186 0.8212 0.8238 0.8264 0.8289 0.8315 0.8340 0.8365 0.8389
1.0 0.8413 0.8438 0.8461 0.8485 0.8508 0.8531 0.8554 0.8577 0.8599 0.8621
1.1 0.8643 0.8665 0.8686 0.8708 0.8729 0.8749 0.8770 0.8790 0.8810 0.8830
1.2 0.8849 0.8869 0.8888 0.8907 0.8925 0.8944 0.8962 0.8980 0.8997 0.9015
1.3 0.9032 0.9049 0.9066 0.9082 0.9099 0.9115 0.9131 0.9147 0.9162 0.9177
1.4 0.9192 0.9207 0.9222 0.9236 0.9251 0.9265 0.9279 0.9292 0.9306 0.9319
1.5 0.9332 0.9345 0.9357 0.9370 0.9382 0.9394 0.9406 0.9418 0.9429 0.9441
1.6 0.9452 0.9463 0.9474 0.9484 0.9495 0.9505 0.9515 0.9525 0.9535 0.9545
1.7 0.9554 0.9564 0.9573 0.9582 0.9591 0.9599 0.9608 0.9616 0.9625 0.9633
1.8 0.9641 0.9649 0.9656 0.9664 0.9671 0.9678 0.9686 0.9693 0.9699 0.9706
1.9 0.9713 0.9719 0.9726 0.9732 0.9738 0.9744 0.9750 0.9756 0.9761 0.9767
2.0 0.9772 0.9778 0.9783 0.9788 0.9793 0.9798 0.9803 0.9808 0.9812 0.9817
2.1 0.9821 0.9826 0.9830 0.9834 0.9838 0.9842 0.9846 0.9850 0.9854 0.9857
2.2 0.9861 0.9864 0.9868 0.9871 0.9875 0.9878 0.9881 0.9884 0.9887 0.9890
2.3 0.9893 0.9896 0.9898 0.9901 0.9904 0.9906 0.9909 0.9911 0.9913 0.9916
2.4 0.9918 0.9920 0.9922 0.9925 0.9927 0.9929 0.9931 0.9932 0.9934 0.9936
2.5 0.9938 0.9940 0.9941 0.9943 0.9945 0.9946 0.9948 0.9949 0.9951 0.9952
2.6 0.9953 0.9955 0.9956 0.9957 0.9959 0.9960 0.9961 0.9962 0.9963 0.9964
2.7 0.9965 0.9966 0.9967 0.9968 0.9969 0.9970 0.9971 0.9972 0.9973 0.9974
2.8 0.9974 0.9975 0.9976 0.9977 0.9977 0.9978 0.9979 0.9979 0.9980 0.9981
2.9 0.9981 0.9982 0.9982 0.9983 0.9984 0.9984 0.9985 0.9985 0.9986 0.9986
3.0 0.9987 0.9987 0.9987 0.9988 0.9988 0.9989 0.9989 0.9989 0.9990 0.9990
3.1 0.9990 0.9991 0.9991 0.9991 0.9992 0.9992 0.9992 0.9992 0.9993 0.9993
3.2 0.9993 0.9993 0.9994 0.9994 0.9994 0.9994 0.9994 0.9995 0.9995 0.9995
3.3 0.9995 0.9995 0.9995 0.9996 0.9996 0.9996 0.9996 0.9996 0.9996 0.9997
3.4 0.9997 0.9997 0.9997 0.9997 0.9997 0.9997 0.9997 0.9997 0.9997 0.9998
a
Enter table by Z-value to obtain cumulative area entry. As an example, area for Z ¼ 1.96 (entry at row 1.9, column 0.06) is 0.9750, indicating
that 97.5% of the standard normal distribution is below this Z-value, and 2.5% is above. Areas for negative Z-values are calculated by subtracting
the area for the positive Z-value from 1. For example, the area for Z ¼ 1.96 is calculated as 1  0.9750 or 0.0250.
b
Table values generated using the SAS System.

exceeds that of the critical upper tail, chi-square value The chi-square table (Table 5) is entered according
at a ¼ 0.05 (p ¼ 0.05 testing level), we reject the to the significance level (e.g., p ¼ 0.05) and the
hypothesis that the sample distribution is consistent degrees of freedom (df) for the calculated statistic.
with the assumed statistical distribution. If our calcu- The degrees of freedom for the goodness-of-fit test
lated value is less than the critical value, we accept are the total number of intervals less one. If, as in
the hypothesis. our case, population parameters for the assumed
 3488 Statistical Methods
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Table 3 Distribution of results of quality value of 170 ng/ml consistent with the assumption
control specimen analyses (hypothesis) that the assay is functioning properly with
Z units Expected Observed only random errors operative. To test this, we calculate
about mean percent percent the Z-statistic, which is equal to (170  207.6)/
3.0 99.7 100 14.1 ¼ 37.6/14.1 ¼ 2.77. From the table of stan-
dard normal deviates, we see that the proportion of
2.5 99 98
the normal distribution that lies within the range 1
2.0 95 93 to 2.77 (or similarly, between 2.77 and þ1) is
1.5 87 88 0.0028. Therefore, the probability of encountering a
1.0 68 74 value this far removed from the mean (37.6 ng/ml)
0.7 52 64 is p ¼ 2  0.0028 ¼ 0.0056. Typically, when there
0.5 38 55 is less than a 5% probability (p < 0.05) of observing
a value, we would question whether our hypothesis
0.2 18 24
was correct. We conclude, therefore, that either we
0.1 8 12 have just observed a relatively rare event or that the
assay was not working as expected. The hypothesis
that the observed value deviates from its expected
statistical distribution are not known and must be esti- value owing only to random fluctuation is called the
mated from the sample, then the degrees of freedom null hypothesis. If the null hypothesis is rejected, then
are further reduced by the number of parameters we accept some stated alternative hypothesis. In this
estimated. In our example using eight intervals and example, the alternative hypothesis is that the assay
estimating the mean and variance of the normal distri- method was not functioning properly.
bution from the sample, there are 8  1  2 ¼ 5 The Z-test can also be used to test if the mean x

degrees of freedom. The critical chi-square value at from a sample of size n is consistent with the known
a ¼ 0.05 with 5 df is 9.24. As Table 4 shows, our mean m of the population. p The Z-statisticp for this test
calculated statistic value, 6.33, is less than the critical is equal to (x
 m)/(s/ n), where s/ n is the stan-
value. Accordingly, we accept the assumption that dard error of the mean. The statistic is evaluated
the QC values appear to come from the assumed against the table of standard normal deviates just as
normal distribution with m ¼ 207.6 and s ¼ 14.1. we did in determining whether the QC value of
Normal distribution theory can be used to test 170 ng/ml was acceptable.
whether a particular sample value is consistent with It should be noted that although it is common to use
other values or with our past experience. If the mean the 5% level of significance (p < 0.05) for testing, this
m and the variance s2 are known, then we can deter- level is not always appropriate. If the rejection of the
mine how deviant an observed value xi, appears to be 170 ng/ml QC value causes us to evaluate if the con-
by calculating the statistic Z ¼ (xi  m)/s and com- duct and performance of the assay for a particular
paring this with the table ofstandard normal deviates. batch of specimens were correct, then the 5% level is
Suppose that one of the values for our QC specimen probably appropriate. At this level, we would expect
was 170 ng/ml. Past experience has led us to believe to have to investigate theperformance of 1 in 20 of
that the results for this QC specimen are normally the batch runs even when everything was functioning
distributed with m ¼ 207.6 ng/ml and s ¼ 14.1. Is the properly. This would result in what is statistically

Table 4 Chi-square goodness-of-fit test

Theoretical
Zlow Zhigh proportion x
 Zlow  s x
þ Zhigh  s Expected (E) Observed (O) (O  E)2/E
4.00 1.18 0.1190 151.2 191.0 5 5 0.000
1.18 0.72 0.1168 191.0 197.5 5 3 0.800
0.72 0.37 0.1199 197.5 202.4 5 4 0.200
0.37 0.00 0.1443 202.4 207.6 6 7 0.167
0.00 0.37 0.1443 207.6 212.9 6 7 0.167
0.37 0.72 0.1199 212.9 217.8 5 9 3.200
0.72 1.18 0.1168 217.8 224.3 5 2 1.800
1.18 4.00 0.1190 224.3 264.1 5 5 0.000
Sum ¼ 6.33; critical w2 ¼ 9.24.
Statistical Methods 3489

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Table 5 Critical values of the Chi-square and student’s t THE DISTRIBUTION OF THE SAMPLE MEAN
distributionsa
Chi-square values for t Values for We seldom know the population variance and, there-
one-sided test one-sided test fore, must estimate it from a sample. If the population
is normally distributed or if the sample is a large one
DF 0.05 0.025 0.005 0.05 0.025 0.005 from a population that is not normal, then we can con-
1 2.71 3.84 5.02 6.31 12.71 63.69 struct a test to determine whether the sample mean is
2 4.61 5.99 7.38 2.92 4.30 9.92 consistent with the known, or assumed, mean. To
3 6.25 7.81 9.35 2.35 3.18 5.84 do so, we rely on a statistic based on Student’s t-
distribution. The t-distribution, attributed to W.S.
4 7.78 9.49 11.14 2.13 2.78 4.60
Gossett, who wrote under the pseudonym ‘‘Student,’’
5 9.24 11.07 12.83 2.02 2.57 4.03
describes the behavior of the means for samples taken
6 10.64 12.59 14.45 1.94 2.45 3.71 from a normal distribution. The t-distribution is
7 12.02 14.07 16.02 1.89 2.36 3.50 defined entirely by the sample size n, or more typically
8 13.36 15.51 17.53 1.86 2.31 3.36 by its degrees of freedom n  1. The distribution has a
9 14.68 16.92 19.02 1.83 2.26 3.25 shape similar to that of the standard normal distri-
bution, bell-shaped, but for small samples is lower
10 15.99 18.31 20.48 1.81 2.23 3.17
peaked and broader than the standard normal distri-
11 17.28 19.68 21.92 1.80 2.20 3.11
bution. As the sample size increases, the distribution
12 18.55 21.03 23.34 1.78 2.18 3.05 approaches that of the normal distribution, coinciding
14 21.06 23.68 26.12 1.76 2.14 2.98 with it when the sample size is infinite. The t-statistic is
16 23.54 26.30 28.85 1.75 2.12 2.92 used like the Z-statistic, for testing the consistency of a
18 25.99 28.87 31.53 1.73 2.10 2.88 sample mean with the population mean m. The differ-
ence between the two statistics is that for the t-statistic,
20 28.41 31.41 34.17 1.72 2.09 2.85
s is replaced by its sample estimate, the standard
p devia-
25 34.38 37.65 40.65 1.71 2.06 2.79
tion s. The statistic is t ¼ (x
 m)/(s/ n). Table 5
30 40.26 43.77 46.98 1.70 2.04 2.75 provides a listing of critical t-values (think of them as
35 46.06 49.80 53.20 1.69 2.03 2.72 t deviates) for different degrees of freedom (n  1).
40 51.81 55.76 59.34 1.68 2.02 2.70 Note that at infinite degrees of freedom, the critical
50 63.17 67.50 71.42 1.68 2.01 2.68 t-value is simply the standard normal deviate Z.
Assume that the QC specimen was supposed to be
60 74.40 79.08 83.30 1.67 2.00 2.66
prepared to contain 200 ng/ml of drug and that we
1 — — — 1.645 1.960 2.576
do not know the true population variance. Is a sample
a
Table values generated using the SAS System. mean equal to 207.6 ng/ml based on 42 determinations
consistent with the true value being 200 ng/ml? This
typically would be stated in the form of a null (H0)
and alternative hypothesis (Ha):
known as a Type I error. A 5% level of extra circum-
spection would generally not be a problem. However,
H0 : m ¼ 200 ng=ml
if the rejection of the 170 ng/ml value led to our
dropping the value from estimates of in-process accu- Ha : m 6¼ 200 ng=ml
racy and precision, then it is not generally acceptable
to erroneously exclude 5% of the values, thereby calcu- Because s for the population is unknown, we must
lating estimates on the best 95% of the results. In this estimate it from the sample standardpdeviation s.
case, a 1% level of significance (p < 0.01), or even We calculate t ¼ (207.6  200)/(14.1/ 42) ¼ 3.49.
lower, would be more appropriate. At this lower level Referring to Table 5 and entering it with 40 degrees
of significance, we would reject fewer values that of freedom, the closest-value to the 42  1 ¼ 41
deviated from the expected value simply owing to degrees of freedom for our calculated statistic, we find
random error. Although this appears to be desirable, a critical, one-sided, t-value at the 0.005 level equal to
testing at a lowered level of significance also reduces 2.70. If the calculated t-statistic value exceeds a tabled
our ability to reject values that differ from expected critical value (t > tcrit) or if it is less than the negation
owing to true assay performance problems. This non- of the critical value (t < tcrit), then the null hypoth-
detection of truly aberrant values is referred to as a esis is rejected. In the example, the calculated value
Type II error. Generally, the only way to decrease both 3.49 exceeds the critical value and we reject the hypoth-
Type I and Type 2II errors is to increase the size of esis that the sample comes from a population with
the sample used in the statistical test. amean of 200 ng/ml at the 0.01 level of significance
 3490 Statistical Methods
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(2  0.005). We, instead, accept the alternative populations with the same mean (or from the same
hypothesis that the QC specimen appears to have been population). A paired t-test can be applied when we
prepared to contain a drug concentration different want to determine whether a newly trained analyst per-
than 200 ng/ml. This is an example of a one-sample forms an assay method with the same proficiency as an
t-test. experienced analyst. Suppose that each analyst pro-
Another application of the t-test is the two-sample cesses seven different QC specimens and we obtain
t-test. This test is used to determine whether two sam- the following assay results, in the order (new analyst,
ples come from populations with the same mean experienced analyst): (12.6, 11.3), (3.46, 2.34), (25.4,
(m1  m2 ¼ 0) or whether the population means differ 22.5), (10.3, 8.80), (5.89, 4.68), (16.4, 14.2), and (9.95,
by some hypothesized amount (m1  m2 ¼ c). In both 8.20). The paired t-test deals with the differences for
cases, the statistic iscalculated as t ¼ [(x1  x2)  each of the paired results (new–experienced): 1.3,
(m1  m2)]/se, where x1 and x2 are the means of first 1.12, 2.9, 1.5, 1.21, 2.2, and 1.75. The mean difference
sample and the second sample. The denominator of is 1.71, with a standard deviation of 0.641. The t-stat-
the statistic is the standard error for the difference istic is calculated by dividingpthis mean by the standard
between the two means, calculated as: se ¼ sp(1/ error (standard deviation/ n) and comparing the
n1 þ 1/n2)2, where, sp ¼ [((n1  1)s21 þ (n2  1)s22)/ result with the critical t-value with n  1 degrees of
(n1 þ n2  2)]2, s1 and n1 are the standard deviation freedom. With the n ¼ 7 pairs in the p example, the cal-
and size of the first sample, and s2 and n2 are the stan- culated t-statistic is 1.71/(0.641/ 7) ¼ 7.06. This
dard deviation and size of the second sample. We enter exceeds the critical t-value 2.45 for the two-sided,
the t-table with n1 þ n2  2 degrees of freedom. If 0.05 level of significance (the one-sided 0.025 value in
the tabled value is exceeded by our calculated value Table 5) with six degrees of freedom. We conclude that
or if the calculated value is less than the negation of a difference as large as the one observed between the
the tabled value, then we reject the hypothesis that two analysts would not likely be attributable to ran-
the two samples are from populations with the same dom error alone. The ‘‘new’’ analyst may need some
mean or differ by the hypothesized amount c. additional training.
Assume that we have a second set of 20 QC values
from a second QC specimen, presumably prepared
identically to the specimen from our previous set of ANOTHER USEFUL DISTRIBUTION
values, and found a mean of 199.6 ng/ml and standard
deviation 13.5 ng/ml. We can evaluate whether these A commonly encountered statistical distribution is the
results are consistent with those from 42 results on binomial distribution. This distribution deals with the
the first QC specimen. The null hypothesis for the test behavior of binary outcomes such as the flip of a coin
is that the two QC specimens have been prepared iden- (heads/tails), the gender of a child (boy/girl), or the
tically, m1  m2 ¼ 0. The alternative hypothesis determination if a tablet has acceptable potency
would be that they are not identical, m1  m2 6¼ 0. (pass/fail). When dealing with a sequence of inde-
The standard error is se ¼ [(41  14.12 þ 19  pendent binary outcomes, such as multiple flips of a
1 1 1
13.5 2 )/(41 þ 19)] =2 (1/42 þ 1/20) =2 ¼ (193.566) =2 coin or determining whether the potencies of 20 tablets
1
=2
(1/42 þ 1/20) ¼ 3.78. are individually acceptable, the binomial distribution
The t-statistic is (207.6  199.6)/3.78 ¼ 2.12. can be used. The probability of observing x successes
Entering Table 5 with 60 degrees of freedom, we obtain in n outcomes is C(x,n) pxqnx. Binomial expansion
a critical t-value for a one-sided test at the 0.025 level for x ¼ 1 to n is C(0,n)p0qn þ C(1,n)p1qn1 þ
of significance equal to 2.00. As the alternative hypoth- C(2,n)p2qn2 þ


þ C(n,n)pnq0. This sum equals 1,
esis is that the two QC specimens differ, without regard as the probability of observing at least one of the pos-
to which has the higher and which has the lower value, sible outcomes is 1 (a certainty). The notation in the
the test is a two-sided test. The critical table value at expansion C(a,n) is the number of ways of obtaining
0.025, one-sided, is the critical value to use for a two- groups of size a from n distinct items: n!/[a!(n  a)!].
sided test at 0.05. The calculated statistic exceeds this As an example, the number of groups of size
critical value, and we reject the hypothesis that the three obtainable when there are four different items
two sets of QC samples were prepared identically. If is C(3,4) ¼ 4!/[3!(4  3)!] ¼ (4  3  2  1)/
we have the stock solutions used to prepare the two [(3  2  1)(1)] ¼ 4. Note that 0! is defined as 1.
QC specimens, we would probably analyze them to The p in the binomial expansion is the success prob-
see whether they have identical concentrations. ability for the single binary event such as observing
The t-distribution can also provide a tool to evalu- a head with one flip of a coin. The q stands for the
ate whether two samples on which paired determina- single-event failure probability (e.g., observing a tail)
tions had been obtained appear to come from and is equal to 1  p.
Statistical Methods 3491

Sterilization
Statistical–
PUTTING IT ALL TOGETHER Applying the binomial expansion, the probability of
at least four of the six QC values passing is
In the analyses of blood specimens from subjects par- C(6,6)p6q0 þ C(5,6)p5q1 þ C(4,6)p4q2 ¼ 1(0.7888)6 þ
ticipating in bioavailability studies, the FDA instructs 6(0.7888)5 (0.2112) þ 15(0.7888)4(0.2112)2, which equals
laboratories to include quality control specimens 0.8869. For a batch to pass, however, we have the
(QC) at each of three known concentrations (low, additional restriction that when only four of six values
mid, and high). The QC specimens are processed in pass, no more than one of the two failures can occur at
duplicate with each batch of subject specimens. The the same QC level (low, mid, or high). This restriction
acceptance criteria for the batch, based on the results reduces the 15 possible ways that four of six QC values
of these QC specimens, is that at least four of the six can pass to 12. This reduces the probability of the batch
values must fall within a specified range about their being accepted to 0.8869  3(0.7888)4(0.2112)2 or
nominal concentrations. In addition, no more than 0.8351. There is an 83.5% probability of accepting a batch
one value at each of the three QC concentration levels run, and there is nearly a 16.5% chance of rejecting it
can be outside its acceptance range. Combining because of random error. We might consider improving
binomial and normal distribution theory, we can esti- the precision of the assay rather than proceeding with a
mate the number of batch runs we expect to reject method that is anticipated to erroneously reject nearly
because of random error. 17% of our analyses. If the CV% of the method were
Assume that the acceptance limit for each QC improved to 8%, the probability of batch acceptance
value is that it must fall within 15% of its nominal increases to 0.9875, and our expected failure rate is
concentration. Any value meeting this criterion passes only 1.2%. With 42 batch runs, we expect to have to
and any not meeting this criterion fails (a binary out- repeat 1–2 batches, as contrasted with the 12% CV assay,
come). The probability of accepting a batch run where we expect to repeat seven batches. Here is a case in
based on multiple binary outcomes (QC pass/fail which a modest improvement in assay precision reaps
determinations) will be governed by the binomial dis- big rewards.
tribution. The probability of a single QC value failing The chi-square distribution used in the goodness-of-
is equal to the probability of obtaining a concen- fit test is useful in another important statistical test.
tration outside 85–115% of its nominal concentration. Assume that previous experience leads us to believe
The concentration estimates are assumed to be nor- that the s of an assay method is no greater than
mally distributed with a mean equal to 100% of the 10 ng/ml. In processing 42 batches, the observed
nominal concentration. The sigma value for the esti- standard deviation was 14.1 ng/ml for the QC values.
mates is equal to the CV of the assay. If the assay Is this result consistent with the prior knowledge of
CV is 12%, the probability of a QC value passing the assay (H0: s  10), or does it appear that the pre-
acceptance criteria is the probability of obtaining a cision of the assay has deteriorated (Ha:s > 10)?
Z-value between (85  100)/12 and (115  100)/12 Application of a chi-square test can help answer this
or between 1.25 and þ1.25. The proportion of the question. The appropriate statistic in this caseis
standard normal distribution between 1.25 and w2 ¼ (n  1)s2/s2, which follows a chi-square distri-
þ1.25 is 0.7888, which is p, the probability of a single bution with n  1 degrees of freedom. In our
QC value passing. The value of q is 1  0.7888 or example, we replace s in the statistic with 14.1, our
0.2112. There are three mutually exclusive ways that sample standard deviation, and s with our assumed
at least four of the six QC values can pass the accept- upper limit of 10 for s. The calculated statistic for this
ance criterion. All six values could pass, five of the one-sided test is w2 ¼ (42  1)14.12/102 ¼ 81.51.
six could pass, or four of the six could pass. These This calculated value exceeds the critical chi-square
outcomes are mutually exclusive; the occurrence of value 59.34 at the 0.005 level of significance and 40
any one of them excludes the possibility that either (approximately 41) degrees of freedom. We conclude
of the other two occurs. The probability of at least that the assay precision had deteriorated and would
four of six QC values passing, then, is the sum of probably launch an investigation as to why.
the probabilities of each of the three mutually exclus- In manufacturing a drug product, it is common to
ive ways in which this event can occur. It should be collect and analyze specimens from the mix of active
noted that in probability calculations, when an event and inactive ingredients, the blend. This is done to ver-
A can occur through any one of the mutually ify adequate uniformity of the blend before proceeding
exclusive events B, C, D, etc., then the probability with the manufacturing process. The specimens are
of A is the sum of the individual probabilities of B, strategically collected from the blend container, for
C, D, etc. However, if A occurs only when events example, from the left, center, and right regions of
B, C, D, etc. all occur, then the probability of A is the top, middle, and bottom of the container. Blend
the product of the individual probabilities of B ,C, uniformity criteria usually require that the mean assay
D, etc. value for the specimens falls within an acceptable range
 3492 Statistical Methods
Sterilization
Statistical–

(e.g., 95–105%) about the label claim (100% potency) approximately 1 in 30, based on using the mean to 1
for the product. In addition, the relative standard devi- in 11 based on using individual values, despite the
ation CV% for the analyses of the blend specimens widened acceptance limits.
must not be greater than some specified limit (e.g.,
5%). Using the characteristics of the normal and the
chi-square distributions, we can estimate the chances ONE FINAL DISTRIBUTION
of passing blend uniformity criteria.
Suppose that prior knowledge of the manufacturing There is one final statistical distribution, the F-
process leads us to believe that we routinely produce distribution, that is an important addition to the basic
blends with potencies between 97 and 103% of label statistical tool chest. This distribution is used in the
claim. We estimate that the variability for a ‘‘good’’ evaluation of two variance estimates to determine
blend is no more than 3% CV, a composite of true whether they are consistent with each other. The QC
blend inhomogeneity, sampling error, and analytical sample based on 42 estimates (41 degrees of freedom)
method variability. What is the probability of accept- had a standard deviation of 14.1 ng/ml. If we had
ing a good blend? The standard error of the mean another set of 31 QC values (30 degrees of freedom),
for nine specimens from the blend will be 1%, our perhaps from a second bioavailability study, with a
worst-case estimate of CV for a single determination standard deviation of 19.5 ng/ml, we might want to
(3%) divided by the square root of the number of speci- know whether the assay precision values for the two
mens collected (9). Assuming that the true potency of studies were consistent. The variance ratio statistic is
the blend is 97% (a worst-case estimate), then the prob- s21/s22, where s1 is the higher of the two standard devia-
ability of observing a mean from nine blend specimens tions, and s2 is the lower of the two. The calculated
that is within our acceptance range of 95–105% is the value of the statistic is compared with tables of critical
same as the probability of observing a Z-value between F-values with n1  1 numerator and n2 1 denomi-
(95  97)/1 and (105  97)/1 or between 2 and nator degrees of freedom. In our case, the critical
þ8. The proportion of the standard normal distri- F30,41 is approximately equal to 1.74 at the 5% level
bution contained between 2 and þ8 is 0.977, the of significance (Table 6 for F30,40, the closest value).
probability of passing the first criterion for blend The calculated ratio is 19.52/14.12 ¼ 1.91, which
acceptance. The second criterion is that the CV of exceeds the critical value. The interpretation is that
the nine blend specimens must be less than 5%. With the assay precision for the second of the two studies
a true CV of 3%, this is the probability of observing differs significantly (p < 0.05) from that of the first
a chi-square value less than (n  1)52/32 ¼ (8)(25)/ (it has less precision, greater CV%).
9 ¼ 22.2. With eight degrees of freedom, the prob- The F-distribution has great utility in a statistical
ability of a chi-square value less than 22.2 is approxi- test referred to as analysis of variance (ANOVA).
mately 0.996, which is the probability of passing the ANOVA is a powerful tool for testing the equivalence
second criterion. The probability of accepting the of means from samples obtained from normally dis-
blend material is the product of the individual prob- tributed, or approximately normally distributed, popu-
abilities of passing criterion 1 and criterion 2, or lations. As an example, suppose that the following are
0.977  0.996, which equals 0.973. Therefore, 97% the content uniformity values on 20 tablets from each
of our blend batches are expected to pass if only ran- of four different lots: lot A mean ¼ 99.5%, standard
dom error is operative. deviation ¼ 2.6%; lot B mean ¼ 100.2%, standard
Some have proposed widening the acceptance range deviation ¼ 2.8%; lot C mean ¼ 90.5%, standard
(e.g., 90–110%) but requiring that all individual blend deviation ¼ 2.1%; and lot D mean ¼ 100.3%,
specimens fall within the widened range. Is this an standard deviation ¼ 2.7%.
easier or harder criterion to meet? The probability of Are any of the lots different from the other lots? To
a single-blend specimen being acceptable, is the prob- answer this question, we need to conduct a statistical
ability of the standard normal deviate Z being between test with a null hypothesis H0: m1 ¼ m2 ¼
(90  97)/3 and (110  97)/3 or between 2.33 and m3 ¼ m4 ¼ m (all means are equal to some unknown
þ4.33. This probability is 0.99. For the blend to pass m), and an alternative hypothesis Ha: m1 6¼ m or
the first criterion, the first specimen, the second, and m2 6¼ m or m3 6¼ m or m4 6¼ m (at least one mean is
the third, up through the ninth, must all, independently, not equal to at least one of the other means). It is
be acceptable. The probability of passing this first assumed that all lots have the same, unknown, vari-
criterion becomes 0.999, or 0.914. This, when combined ance, s2. The F-statistic involves the calculation of
with the probability of meeting the second criterion, two variance estimates s21 ¼ 1/(k  1) Snj(x
j  x..)2
results in a probability of blend acceptance being and s22 ¼ 1/(N  k) SS(xij  x
j)2. In the statistic,
0.914  0.996 or 0.91. Only 91% of the blends are x.. is the grand mean across all k ¼ 4 lots, nj is the
expected to pass. The chance failure rate goes from number of values from which the jth lot mean x
j was
Table 6 Critical values of the F-distribution for 5% level of significancea
Numerator degrees of freedom:
Statistical Methods

Dfb 1 2 3 4 5 6 7 8 9 10 11 12 15 20 25 30 40 60 100
1 161 200 216 225 230 234 237 239 241 242 243 244 246 248 249 250 251 252 253
2 18.5 19.0 19.2 19.3 19.3 19.3 19.4 19.4 19.4 19.4 19.4 19.4 19.4 19.5 19.5 19.5 19.5 19.5 19.5
3 10.1 9.55 9.28 9.12 9.01 8.94 8.89 8.85 8.81 8.79 8.76 8.74 8.70 8.66 8.63 8.62 8.59 8.57 8.55
4 7.71 6.94 6.59 6.39 6.26 6.16 6.09 6.04 6.00 5.96 5.94 5.91 5.86 5.80 5.77 5.75 5.72 5.69 5.66
5 6.61 5.79 5.41 5.19 5.05 4.95 4.88 4.82 4.77 4.74 4.70 4.68 4.62 4.56 4.52 4.50 4.46 4.43 4.41
6 5.99 5.14 4.76 4.53 4.39 4.28 4.21 4.15 4.10 4.06 4.03 4.00 3.94 3.87 3.83 3.81 3.77 3.74 3.71
7 5.59 4.74 4.35 4.12 3.97 3.87 3.79 3.73 3.68 3.64 3.60 3.57 3.51 3.44 3.40 3.38 3.34 3.30 3.27
8 5.32 4.46 4.07 3.84 3.69 3.58 3.50 3.44 3.39 3.35 3.31 3.28 3.22 3.15 3.11 3.08 3.04 3.01 2.97
9 5.12 4.26 3.86 3.63 3.48 3.37 3.29 3.23 3.18 3.14 3.10 3.07 3.01 2.94 2.89 2.86 2.83 2.79 2.76
10 4.96 4.10 3.71 3.48 3.33 3.22 3.14 3.07 3.02 2.98 2.94 2.91 2.85 2.77 2.73 2.70 2.66 2.62 2.59
11 4.84 3.98 3.59 3.36 3.20 3.09 3.01 2.95 2.90 2.85 2.82 2.79 2.72 2.65 2.60 2.57 2.53 2.49 2.46
12 4.75 3.89 3.49 3.26 3.11 3.00 2.91 2.85 2.80 2.75 2.72 2.69 2.62 2.54 2.50 2.47 2.43 2.38 2.35
13 4.67 3.81 3.41 3.18 3.03 2.92 2.83 2.77 2.71 2.67 2.63 2.60 2.53 2.46 2.41 2.38 2.34 2.30 2.26
14 4.60 3.74 3.34 3.11 2.96 2.85 2.76 2.70 2.65 2.60 2.57 2.53 2.46 2.39 2.34 2.31 2.27 2.22 2.19
15 4.54 3.68 3.29 3.06 2.90 2.79 2.71 2.64 2.59 2.54 2.51 2.48 2.40 2.33 2.28 2.25 2.20 2.16 2.12
16 4.49 3.63 3.24 3.01 2.85 2.74 2.66 2.59 2.54 2.49 2.46 2.42 2.35 2.28 2.23 2.19 2.15 2.11 2.07
17 4.45 3.59 3.20 2.96 2.81 2.70 2.61 2.55 2.49 2.45 2.41 2.38 2.31 2.23 2.18 2.15 2.10 2.06 2.02
18 4.41 3.55 3.16 2.93 2.77 2.66 2.58 2.51 2.46 2.41 2.37 2.34 2.27 2.19 2.14 2.11 2.06 2.02 1.98
20 4.35 3.49 3.10 2.87 2.71 2.60 2.51 2.45 2.39 2.35 2.31 2.28 2.20 2.12 2.07 2.04 1.99 1.95 1.91
30 4.17 3.32 2.92 2.69 2.53 2.42 2.33 2.27 2.21 2.16 2.13 2.09 2.01 1.93 1.88 1.84 1.79 1.74 1.70
40 4.08 3.23 2.84 2.61 2.45 2.34 2.25 2.18 2.12 2.08 2.04 2.00 1.92 1.84 1.78 1.74 1.69 1.64 1.59
50 4.03 3.18 2.79 2.56 2.40 2.29 2.20 2.13 2.07 2.03 1.99 1.95 1.87 1.78 1.73 1.69 1.63 1.58 1.52
60 4.00 3.15 2.76 2.53 2.37 2.25 2.17 2.10 2.04 1.99 1.95 1.92 1.84 1.75 1.69 1.65 1.59 1.53 1.48
100 3.94 3.09 2.70 2.46 2.31 2.19 2.10 2.03 1.97 1.93 1.89 1.85 1.77 1.68 1.62 1.57 1.52 1.45 1.39
1 3.84 3.00 2.60 2.37 2.21 2.10 2.01 1.94 1.88 1.83 1.79 1.75 1.67 1.57 1.51 1.46 1.39 1.32 1.24
a
Df are the denominator degrees of freedom.
b
Table values generated using the SAS System.
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determined (20 for each lot), and N ¼ Snj ¼ SSnij etc., then for each of these 5% level tests, we have a
is the total number of xij values (20 þ 20 þ 95% probability of obtaining a correct assessment
20 þ 20 ¼ 80). The s21 is a pooled variance estimate when the null hypothesis that all means are equal is
of how the category means vary about the grand mean, true. To correctly accept thenull hypothesis of all
and s22 is a pooled variance estimate of how the individ- means being equal, we must simultaneously conclude
ual values within each category vary about their that A ¼ B and B ¼ C and C ¼ D and A ¼ C,
category mean. If the identification (grouping) of the etc., from the multiple t-tests . The probability of doing
values into categories (lots) does not affect the variance so is (0.95)(0.95)(0.95)(0.95) ¼ (0.95)n, where n is the
estimate, then the variance ratio s21/s22 will differ from number of pair-wise comparisons conducted by t-tests.
1.0 by only random error. The F-distribution describes The probability of being incorrect in this multiple pair-
how the ratio varies about 1.0 owing to random error wise approach is 1  (0.95)n, which exceeds the
when a set of values are arbitrarily grouped into desired 5% level for any n > 1. In our case, there
categories. If the calculated statistic value exceeds the are C(4,2) or six possible pair-wise comparisons. The-
critical Fk1, Nk tabled value, then the null hypothesis multiple pair-wise approach has a probability of an
of equal means is rejected. incorrect assessment, Type I error when the null
In our example, x
¼ Snjx
j/N ¼ (20  99.5 þ hypothesis is true, equal to 1  (0.96)6 ¼ 0.265. This
20  100.2 þ 20  90.5 þ 20  100.3)/80 ¼ 97.625. approach would essentially be testing at a 26.5% level
s 21 ¼ (1/3)[(99.5  97.625)2 þ (100.2  97.625)2 þ of significance, rather than at the desired 5% level.
(90.5  97.625) 2 þ (100.3  97.625) 2 ] ¼ 22.689. For this reason, we only consider pair-wise examina-
Because the square of the standard deviation in each tion of the data when the global assessment of equality
lot j is s2j ¼ S(xij  x
j)2/(nj  1), we can calculate of means is rejected by ANOVA. This maintains the
s22 as (1/(N  k))S(nj  1) s2j ¼ 1/(80  4) desired 5% significance level.
[19  2.6 þ 19  2.8 þ 19  2.1 þ 19  2.72] ¼
2 2 2
For the post-ANOVA, pair-wise evaluations, there
69.0. The calculated variance ratio is 53.8/ are procedures to deal with the multiple comparison
6.575 ¼ 3.45 and the critical 5% level F3,76 is <2.76. problem. One such procedure is based on the
The critical value 2.76 is that of F3,60 in Table 6. This F-distribution with one and N  k degrees of freedom.
value is the closest tabled value with the desired numer- This test also relies on the value of s22 from ANOVA.
ator degrees of freedom (3) and denominator degrees of The test statistic is F ¼ (1/s22) [(x.1  x.2)2/(1/
freedom 60, which do not exceed N  k ¼ 76. As the n1 þ 1/n2)], where x.1, x.2 are the means of the n1
calculated statistic value exceeds the critical F-value, we and n2 values for the two lots in the pair-wise compari-
reject the null hypothesis that the means of all lots son. Comparing lot A and lot C: F ¼ (1/6.575)
are the same. We accept the alternative hypothesis [(99.5  90.5)2/(1/20 þ 1/20)] ¼ 123.2. This far
that at least one of the four means differs statistically exceeds the critical F1,76 value at even a 1% level, which
from at least one of the other means. Looking at is <7.08, based on F1,60, and we therefore reject the
the four means, it appears that lot C differs from hypothesis that lot A and lot C means are equal.
the others. Because, the means for lot B and lot D differ from that
Instead of performing ANOVA, we might have con- of lot C by an even greater amount, they also are found to
sidered pairing each lot mean with each of the other lot be statistically different from the lot C mean. By contrast,
means and performing multiple two-sample t-tests to the comparison of lots A and D, with means of 99.5%
determine which ones differ significantly from the and 100.3%, respectively, have an F-test value of 0.97,
others. This approach would not be acceptable. First, far less than the critical 5% value, which is <4.00.
it does not use all of the available information for The test of the four lots is an example of a one-way
the pooled denominator variance estimate. Second, it ANOVA. The one-way comes from the fact that there
introduces what is known as the multiple comparison is only one category (lot) into which the data is classi-
problem. The problem arises if there are multiple, sep- fied. Often, we have more than one category (class vari-
arate, statistical evaluations conducted on the same set able) in which we need to classify data. Although our
of data. When we conduct a single test of a hypothesis interest may be to determine only whether a particular
at the 5% level of significance (a ¼ 0.05), such as H0: class variable has meaning, it is important to include
A ¼ B, we expect to falsely reject the hypothesis 5% other class variables that may influence the variability
of the time when A actually equals B (Type I error). of the data. ANOVA involves a null hypothesis for
We have, therefore, a 95% probability of being correct each classification variable that proposes that the
in the assessment when the null hypothesis A ¼ B means at each different level of the class (category)
is true. This is also true for the F-test of are all equal. If we reject the null hypothesis we con-
A ¼ B ¼ C ¼ D in ANOVA. If we were to indepen- clude in favor of the alternative hypothesis, that at
dently perform multiple t-tests on our data with least one mean in the class differs from at least one
hypotheses such as A ¼ B, B ¼ C, C ¼ D, A ¼ C, other mean in the class. This is also a conclusion that
Statistical Methods 3495

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categorizing the data by that class variable has Table 7 Evaluation of the proficiency of two
meaning. analysts to process plasma samples
Assume that we have two analysts to determine the Specimen Day Analyst 1 Analyst 2
drug concentrations in plasma specimens from a bio- A 1 52.8 57.9
availability study, using our previously described ana-
A 1 52.3 57.5
lytical method. Before starting the analyses, we want to
determine whether they are equally proficient with the A 2 53.3 58.0
method. We might set up the test by obtaining plasma A 2 52.1 57.0
specimens at three different concentrations of drug. B 1 42.2 45.6
We then have each analyst process in duplicate each B 1 42.4 44.2
specimen on each of two different days. The resulting B 2 41.7 43.8
drug concentration values can be categorized by three
B 2 40.4 44.1
different class variables: analyst, day, and specimen.
The variable analyst has two possible levels, analysts C 1 56.8 64.0
one and two. Day has two levels (days 1 and 2), C 1 57.0 64.9
and specimen has three (A, B, and C). ANOVA on C 2 59.6 62.6
the results is a three-way one, named for the three C 2 57.2 61.8
classification variables included. Our interest is in the
classification by analyst, but the other two variables
are necessary to properly define how the experiment variable plus random error: SSTotal ¼ SSAnalyst þ
was conducted. Table 7 shows the results of the SSDay þ SSSpecimen þ SSError. The total sums of
experiment. squares are calculated as the difference between two
ANOVA calculations are straightforward in this ANOVA quantities, A  C. The sums of squares for
example and are easily expanded to situations in which each classification variable is its ANOVA quantity
there are higher numbers of categories. The first minus C. The sums of squares for random error is
ANOVA quantity we calculate is C, the correction fac- determined by difference: SSError ¼ SSTotal 
tor. C is simply the square of the sum of all the individ- SSAnalyst  SSDay  SSSpecimen. The degrees of free-
ual values, divided by N, the total number of values: dom are also additive with dfTotal ¼ dfAnalyst þ
(Sxijkl)2/N, where i ¼ 1  2 (analysts), j ¼ 1  2 dfDay þ dfSpecimen þ dfError. The total degrees of
(days), k ¼ 1  3 (specimens), and l ¼ 1  2 freedom are simply the total number of values less
values for each specimen for each analyst on each one. The degrees of freedom for each class variable
day. Table 8 demonstrates the calculation of this and are the number of levels within the class less one.
the other calculated ANOVA quantities. The quantity The error degrees of freedom are determined by differ-
A is calculated as the sum of the squared individual ence. Dividing each sum of squares by its degrees of
values: Sx2ijkl. The ANOVA quantity for each classi- freedom provides the mean square MS, which is an
fication variable (category) is the sum of the squared estimate of the population variance s2.
totals for each level of the classification, divided by Table 9 shows the construction of the ANOVA
the number of values in each level of the classification. table. If the variance estimate of a class variable
As seen with the variable specimen, this is the sum of MSVariable deviates significantly from that obtained by
the squared totals for the values for each of the speci- that for random error MSError, then the null hypothesis
mens A, B, and C, all divided by 8, the number of that the means at the different levels for that variable
values for each specimen (2 values for each of 2 days are equal is rejected. In other words, the classification
for each of 2 analysts). The principle of ANOVA is of data by that variable is explanatory of the variation
that the total sum of squares SSTotal is divisible into observed in the data. We conduct the test by using
its component sums of squares for each classification the variance ratio test F ¼ MSVariable/MSError, with

Table 8 Calculation of ANOVA quantities

Totals No. of observations S (Totals squared)/No.
All 24 values 1269.2 24 C ¼ 67119.53
Days 1 and 2 637.6, 631.6 12 T ¼ 67121.03
Analysts 1 and 2 607.8, 661.4 12 K ¼ 67239.23
Specimens A, B, and C 440.9, 344.4, 483.9 8 R ¼ 68395.42
Square of each value A ¼ 68541.24
 3496 Statistical Methods
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Table 9 Calculation of ANOVA table

ANOVA
Source of variation DF SS MS ¼ SS/DF F-ratio
Day 1 1.5000 (T  C) 1.5000 1.16
Analyst 1 119.7067 (K  C) 119.7067 92.42a
Specimen 2 1275.8958 (R  C) 637.9479 492.51a
Error 19 24.6108 (A  T  K  R þ 2C) 1.2953
Total 23 1421.7133 (A  C)
a
Indicates statistical significance (p < 0.05).

numerator degrees of freedom equal to those of the The means for analyst 1 was 607.8/12, or 50.65 and
variable and denominator degrees of freedom equal for analyst 2 was 661.4/12, or 55.12. The difference
to that of the error term. As shown in Table 9, we between the means is 4.47. The standard error se
reject the hypothesis that the two analysts process the for the difference would be(s2p/n1 þ s2p/n2)0.5 ¼
specimens equivalently. We also see, as we expected, (2s2p/n)0.5 ¼ (2MSError/12)0.5 ¼ (2  1.2953/12)0.5 ¼
that the specimen levels A, B, and C are not equal. 0.465. The critical t-value for a 95% confidence interval
There are no detectable differences in the values is the one-sided, 0.025 level valueat 19 degrees of
obtained over the 2 days of processing. freedom, 2.095. The 95% confidence interval is
In the previous example, we tested the hypothesis of (4.47)  (0.465) (2.095) or from 5.44 to 3.50. We
‘‘equality.’’ In reality, our interests are usually not in have 95% confidence that the results of analyst1 are
equality but in ‘‘comparability.’’ We generally do not between 6.34% (100  3.50/55.12) to 9.88%
require, or even logically expect, that two lots of the (100  5.44/55.12) lower than those of analyst 2 using
same pharmaceutical product will have the exact same the assay method. Because the value 0 is not included in
potency. If we had several different lots of a drug pro- the 95% confidence interval, we can conclude at the 5%
duct and analyzed enough units from each (e.g., 100– level that there is a statistically significant difference
200 tablets), we could detect as statistically significant between the two analysts, the same conclusion we
even a 1% difference in the potency between the lots. had with ANOVA. However, a difference of 9.88%,
Although statistically significant, such a difference the maximum confidence interval limit, might not be
would have no practical significance. We require, how- large enough for us to reject that the two analysts are
ever, that the potencies, as with the analysts’ perform- comparable in their performance of the assay method.
ance on an assay, be comparable, allowing for a This is a decision that was not possible based solely on
reasonable margin of error. Although hypothesis test- ANOVA results.
ing deals with tests of equality, the closely related con-
fidence interval approach deals with comparability.
The confidence interval calculation can be applied to
comparisons of means from samples drawn from nor- LINEAR REGRESSION
mal populations or from any population if the samples
are large (thanks to the central limit theorem). First, Those familiar with analytical methods probably have
we calculate the difference between two means from familiarity with ‘‘fitting a straight line’’ through data
samples from the two population x
1  x
2, which esti- points. The statistical method generally used is known
mates the difference between the population means as linear regression or ordinary least-squares. Even the
m1  m2. Next, we need to calculate the standard error simplest scientific calculators and spreadsheet pro-
for this difference. The standard error is calculated as grams contain the methods for determining the slope
se ¼ (s2p/n1 þ s2p/n2)0.5, where n1 and n2 are the num- (m) and intercept (b) of the line relating the dependent
ber of values in the first and second samples, and s2p is a variable y (measured with random error) to the inde-
variance estimate (s2p from t-test and s22 from one-way pendent variable x (without random error),
ANOVA or MSError). We have (1  a)  100% con- y ¼ mx þ b. The statistical form of this is yi ¼
fidence that the true difference between the population mxi þ b þ ei, where ei is the random error for the
means falls within the interval (x
1  x
2)  observation yi at a specific value of xi. The calculations
se  ta/2, where ta/2 is the critical one-sided t-value for fitting the line are easy, with m ¼ [S(xi  x
)
at the a/2 level of significance and degrees of freedom (yi  y
)]/[(xi  x
)2] and b ¼ y
 bx
, where y

equal to those for our variance estimate s2p. and x
are the mean values for all the yi and xi values,
In ANOVA for the results from the two analysts, respectively. It is common to have more than a single y
MSError ¼ 1.2953 with 19 degrees of freedom. value for each x value. A basic assumption is that the
Statistical Methods 3497

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Table 10 Linear regression analyses of calibrator curve data
Calibrator Backa Backb
(ng/ml) Response calculated Accuracy calculated Accuracy
(x) ratio (y) (ng/ml) (%) 1/x (x0 ) y/x (y0 ) (ng/ml) (%)
0.250 0.321 0.134 54 4.000 1.285 0.253 101
0.250 0.311 0.123 49 4.000 1.244 0.242 97
0.500 0.545 0.373 75 2.000 1.090 0.490 98
0.500 0.59 0.419 84 2.000 1.178 0.537 107
1.00 1.042 0.902 90 1.000 1.042 1.02 102
1.00 0.994 0.851 85 1.000 0.994 0.967 97
5.00 4.981 5.10 102 0.200 0.996 5.20 104
5.00 4.466 4.55 91 0.200 0.893 4.66 93
10.0 9.911 10.4 104 0.100 0.991 10.4 104
10.0 9.247 9.65 96 0.100 0.925 9.73 97
25.0 24.132 25.5 102 0.040 0.965 25.5 102
25.0 22.448 23.7 95 0.040 0.898 23.8 95
50.0 44.061 46.7 93 0.020 0.881 46.7 93
50.0 51.171 54.3 109 0.020 1.023 54.3 109
100 101.132 108 108 0.010 1.011 107 107
100 88.314 93.9 94 0.010 0.883 93.7 94
250 227.438 242 97 0.004 0.910 242 97
250 241.472 257 103 0.004 0.966 256 103
a
Slope ¼ 0.938; intercept ¼ 0.195.
b
Slope ¼ 0.941; intercept ¼ 0.0834.

variance (error) in the determination of each yi value is of variance for yi. To correct for this, we can use a
independent of its corresponding xi value. variance-stabilizing transformation. If we divide the
As previously indicated, the determination of a drug equation of the line by xi, we obtain yi/xi ¼
concentrations in plasma specimens requires the con- m þ b(1/xi) þ ei/xi. However, ei/xi ¼ K is a con-
struction of a calibration response curve. This curve stant, resulting in equal variance for yi0 ¼ yi/xi. When
is often constructed as a straight line from the mea- yi0 is regressed against 1/xi, we obtain a line with slope
sured peak response ratios (yi) plotted against their equal to our desired intercept b and an intercept equal
respective calibrator concentrations (xi). The drug con- to the desired slope m. The last four columns of
centration in a specimen or the apparent (back-calcu- Table 10 demonstrate with calibrator data how this
lated) calibrator concentration is obtained from a transformation provides accurate estimates for the back-
rearrangement of theequation for the calibration line calculated concentrations. The variance-stabilized line
(without error) xi ¼ (yi  b)/m. An example of a provides a slope estimate that differs only slightly from
calibration curve with back-calculated concentrations that of the original line (0.938 ! 0.941) and an intercept
is given the first four columns of Table 10. estimate that differs more substantially (0.195 !
It is obvious that the lower end of the calibration 0.0834). The approach of weighted least-squares, avail-
curve does not provide an accurate representation of able in many advanced regression programs, gives the
the calibrator concentrations, a problem that does same results, using a 1/x2 weighting, and provides a
not exist at the higher end of the curve. This problem greater choice for variance stabilization.
illustrates what can happen when there is violation of
the assumption of equal (homogeneous) variance for
the yi values. In bioanalytical methods, the largest CONCLUSIONS
component of the random error can often be attributed
to volume errors, One often finds that the standard The methods presented will hopefully provide a basic
deviation of the response (yi) is proportional to the foundation for the application of statistics to pharma-
concentration (xi), that is, ei ¼ kxi, where k is a ceutical problems. Because of space limitations, the
constant. This violates the assumption of homogeneity discussion has been limited to situations in which
 3498 Statistical Methods
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known statistical distributions could be assumed Brown, B.W., Jr.; Hollander, M. Statistics: A Biomedical Intro-
(parametric analyses). This is not always the case with duction; John Wiley & Sons: New York, 1977.
Chatterjee, S.; Price, B. Regression Analysis by Example; John
real-life data. Fortunately, there are a number of non- Wiley & Sons: New York, 1977.
parametric, distribution-free methods available to the Dixon, W.J.; Massey, F.J., Jr. Introduction to Statistical Analy-
pharmaceutical scientist to deal with analyses of such sis, 3rd Ed.; McGraw-Hill Co.: New York, 1969.
Draper, N.R.; Smith, H. Applied Regression Analysis, 2nd Ed.;
data. A general knowledge of statistical methods is a John Wiley & Sons: New York, 1981.
necessity for the pharmaceutical scientist. It is, how- Kanji, G.K. 100 Statistical Tests; Sage Publications, Inc.:
ever, also important for the scientist to realize when Newbury Park, CA, 1993.
Li, C.C. Introduction to Experimental Statistics; McGraw-Hill
a problem or experimental design requires consultation Co.: New York, 1964.
with a statistician. The pharmaceutical statistician has Matthews, D.E.; Farewell, V.T. Using and Understanding
a toolbox of methods considerably more advanced Medical Statistics, 3rd revised Ed.; Karger AG: Basel,
Switzerland, 1996; 5.
than the few basic methods presented here. Meyer, S.L. Data Analysis for Scientists and Engineers; John
Wiley & Sons: New York, 1975.
Snedecor, G.W.; Cochran, W.G. Statistical Methods, 8th Ed.;
Iowa State University Press: Ames, 1989.
SASÕ System. Base SAS and SAS/STATÕ Software; Release
BIBLIOGRAPHY 6.12 TS060 for Windows; SAS Institute, Inc.: Cary, NC,
1989–1996.
Bolton, S. Pharmaceutical Statistics, 3rd Ed.; Marcel Dekker, Zar, J.H., Ed.; Biostatistical Analysis; Prentice-Hall: Englewood
Inc.: New York, 1997. Cliffs, NJ, 1974.
 Sterilization
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Statistical Process Control and Process Capability
Thomas D. Murphy
Morristown, New Jersey, U.S.A.

Shailesh K. Singh
Merlin L. Utter
Wyeth Research, Pearl River, New York, U.S.A.

INTRODUCTION to any process, such as services, office procedures, and

quality assurance functions. Process capability analy-
Scope sis assesses the ability of the process to consistently
meet specifications.
Statistical process control (SPC) provides a statistical In any process, regardless of how well designed or
approach for evaluating processes and for improving maintained, a certain amount of natural or inherent
the quality of these processes through elimination of variability will exist. This natural variability has been
special causes. When SPC is effectively implemented called a ‘‘stable system of chance causes.’’ A process
within a company, benefits can be derived through a operating with only chance causes is said to be in
reduced cost of manufacture, improved quality, fewer statistical control, and the variation is said to be
troubleshooting crises, and improved relationships because of common causes. Statistical process control
with customers. Process capability is a companion evaluates whether or not a process is in statistical con-
tool—one that can be used once a state of statistical trol with respect to one or more process or product
control is achieved—to assess the performance of a characteristics.
process relative to its product specifications. Process If an upset occurs in a process (indicated by a
capability can be used to determine whether processes spike, sustained shift, or trend in the process mean
are capable of continually operating within their stated or by a change in the process variability), then an
specification limits. investigation is initiated to determine the special cause
These tools have been used successfully in such of the upset. The special cause, when determined, is
industries as the automotive and semiconductor indus- either eliminated or controlled by process adjustment.
tries and have been credited for helping to provide By eliminating or controlling the special causes,
great improvements in quality. They have been used process improvement occurs. Root cause analysis is a
to a lesser extent in the pharmaceutical industry but structured investigation that aims to identify a
do provide the same potential for providing improve- special cause and the actions necessary to eliminate it,
ments in quality. This chapter provides a clear expla- and involves the use of a collection of tools including
nation of these concepts and how they can be used in flow charts, brainstorming, cause-and-effect diagrams,
practice. The relevance of these concepts to the phar- and Pareto analysis, as described in other references.[1,2]
maceutical industry and some of the cautions associa- The control chart is the basic analytical tool of SPC
ted with their use will be discussed. Some examples will and is used for first assessing a process, then for moni-
be provided to aid in the understanding of their use. In toring a process output with respect to on-target con-
addition, some possible reasons why these concepts trol and process variability. A control chart is
have been slow in gaining acceptance within the basically a time plot of a statistic calculated from a
pharmaceutical industry will be discussed. variable associated with a process. This variable may
either be a process variable, such as temperature or
Basic Principles flow rate, or a product variable, such as fill weight or
potency. Examples of statistics are an individual
A process is defined as a set of interrelated or interact- measurement, an average of two or more measure-
ing activities that transform inputs into outputs. ments, a percentage of defective output items, a count
Statistical process control is a set of techniques for of defect occurrences in time or space, or a measure of
improving the quality of its output. Although manu- variation such as a range or standard deviation of two
facturing processes first come to mind, SPC can apply or more measurements.
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-120012018
Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved. 3499
 3500 Statistical Process Control and Process Capability
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Superimposed on this time plot are the upper and Table 1 Control chart factors
lower control limits, traditionally set at a distance of n A2 D3 D4 d2

3 times the standard error (SE) of the statistic from
2 1.880 0 3.267 1.128
the center line or process mean. This controls the risk
of a false alarm at a low level (a chance of 3 of 1000 if 3 1.023 0 2.575 1.693
the distribution is normal). The process is said to be in a 4 0.729 0 2.282 2.059
state of statistical control if the plotted points appear 5 0.577 0 2.114 2.326
to occur in a random pattern and are contained within 6 0.483 0 2.004 2.534
the control limits. The centerline and control limits are 7 0.419 0.076 1.924 2.704
calculated from retrospective data from the process.
8 0.373 0.136 1.864 2.847
9 0.337 0.184 1.816 2.970
STATISTICAL PROCESS CONTROL 10 0.308 0.223 1.777 3.078

The Shewhart Chart

The d2 factor is used to provide an estimate of the
The Shewhart Chart is applicable when the data process standard deviation s^ ¼ R=d2 , where the caret
are numeric and multiple data values are taken at (^) denotes an estimated value. More extensive tables
each time point. For discrete parts processes, multiple of control chart factors are available.[3]
items are sampled from the process at each time
point and measured for one or more characteristics.
These multiple measurements constitute a subgroup. Shewhart Chart Example
The number n of measured items in the subgroup
is the subgroup size. The values of the measured An example of a Shewhart Chart is shown below for a
characteristic in a subgroup are denoted Pn as hypothetical powder fill process. Five vials of product
X1,X2, . . . ,Xn. The subgroup average X ¼ i¼1Xi/n were sampled every hour and the net content of each
is calculated and plotted on the Average Chart (also vial was determined. The Shewhart Chart is shown in
called the X-Bar Chart) as a measure of the process Fig. 1. The Average Chart indicates lack of statistical
average at that time point. The subgroup range control at subgroups 3, 4, 9, 14, and 17. Further study
R ¼ Max(Xi)  Min(Xi) is also calculated and of the Average Chart indicates a possible shift in the
plotted on the Range Chart (also called the R-Chart) process mean at subgroup 12, and the Range Chart
as a measure of the process variation at that time shows an increase at subgroup 6. Subsequent special
point. The two charts are used as a unit and constitute cause investigation determined that the shift in process
the Shewhart Chart.
On each of the component charts, the average value
is represented as a solid line, called the central line Average chart
(CL). For the XPchart, the CL is the average of the 304
k 303
averages X ¼ j¼1 Xj =k, and Pfor the R-Chart, the Xbar
302
Average

k
CL is the average range R ¼ j¼1Rj/k, where k is UCL
301
the number of subgroups. CL
300
Around the CL are the control limits, set at
3 SE 299
LCL

of the statistic being plotted. If the statistic value falls 298

outside the control limits, this is a signal that the pro- 0 5 10 15 20
cess is not in a state of statistical control. Because the Subgroup
standard errors are functions of the process standard
Range chart
deviation s, an estimate of this quantity is necessary. 8
This can be supplied by the average range. The lower 7
6
control limit (LCL) and upper control limit (UCL) 5 R
Range

are calculated as follows: 4 CL

3 UCL
2
Chart CL LCL UCL 1
0
Average X X  A2 R X þ A2 R 0 5 10 15 20
Range R D3R D4R Subgroup

Fig. 1 Shewhart control chart for a hypothetical powder fill

The values of A2, D3, and D4 are given in Table 1. process.
Statistical Process Control and Process Capability 3501

Sterilization
Statistical–
mean was caused by a change to a new feed hopper hour, it would be better to sample them all at one time,
containing powder of a higher bulk density before sub- rather than to sample one bottle every 15 min. Sam-
group 12 (the filling was controlled by volume). A line pling over a longer period of time can capture sources
stoppage before subgroup 6 because of a shift change of variability other than inherent variability.
caused increased variability in the fill weights of the Frequency of sampling depends on the dynamics of
product on startup. the process; that is, it depends on how quickly the pro-
The purpose of SPC is to detect special causes so cess reacts to changes in process inputs and on the con-
that they can be eliminated. In this example, two spe- sequences (costs) of failing to react quickly to
cial causes were uncovered: the change in input powder changes.[4] Another consideration is the testing time
bulk density and the unnecessary line stoppages. turnaround, because a high sampling frequency is not
useful if the test results are not available before the
time that the next sample is taken. Typically, physical
Calculations for the Shewhart Chart
measurements may be made in real time whereas
chemical measurements may require laboratory assay.
This section illustrates the calculations required to gen-
The subgroup size is based on a number of considera-
erate the control chart provided in the preceding
tions: cost and ease of sampling, cost and ease of
example. In this case, the control limits were calculated
measurement, and the need to quickly detect shifts from
from the available line data. In subsequent phases of
target of a given magnitude. A subgroup size of four or
SPC, these control limits would be applied to future
five is the most common, especially for physical measure-
data as they are measured from the process. The
ments, because it assures that the average value is
averages and ranges calculated for each subgroup for
normally distributed if the distribution of the individuals
the current data are in Table 2.
is reasonably symmetric. If testing is lengthy or expen-
The overall average of the k ¼ 20 subgroup
sive, the subgroup size can be reduced to two or three.
averages and the overall average range of the sub-
Large subgroup sizes, such as 10 or 20, are unusual
group ranges were calculated using the formulas
Pk and recommended only in those situations where the
X ¼
P i¼1 Xi =k ¼ 6017:6=20 ¼ 300:9 and R ¼ control chart must be sensitive to small shifts in the
k
i¼1Ri/k ¼ 51.6/20 ¼ 2.6. These values were process mean and the testing is rapid and inexpensive.
plotted as the centerlines of the Average and Range The chart in Table 3 indicates how quickly the
Charts, respectively. X-Bar Chart would react to a shift of h standard devia-
For a subgroup size of n ¼ 5, the control chart tions (hs), in terms of the average number of sub-
factors taken from Table 1 were: groups or average run length (ARL), as a function of
subgroup size n.[1] The larger the subgroup size is,
n A2 D3 D4 d2 the more sensitive the chart is to a given shift.
5 0.577 0 2.114 2.326

Supplemental Statistical Process Control Rules

The control chart limits for the Average Chart were: for Detecting Lack of Statistical Control

LCL ¼ X  A2 R ¼ 300:9  ð0:577Þð2:6Þ ¼ 299:4 The process is in a state of statistical control if the values
UCL ¼ X þ A2 R ¼ 300:9 þ ð0:577Þð2:6Þ ¼ 302:4 are within the control limits and the pattern is random.
To help detect special causes, a set of supplemental rules
The control chart limits for the Range Chart were: is available from the ‘‘Statistical Quality Control Hand-
book,’’[5] and a subset of these rules is given below.
LCL ¼ D3 R ¼ ð0Þð2:6Þ ¼ 0 The process is deemed ‘‘out of control’’ when:
UCL ¼ D4 R ¼ ð2:114Þð2:6Þ ¼ 5:5
2 of 3 points fall outside the warning limits (shift)
An estimate of the process standard deviation was 8 points in a row fall above or below the CL (shift)
^ ¼ R=d2 ¼ 2:6=2:326 ¼ 1:1.
s
6 points in a row are steadily increasing or decreas-
ing (trend)
Control Chart Design 14 points in a row alternate up and down (two feed
sources or overadjustment).
A basic concept in the Shewhart Chart methodology is
the division of observations into rational subgroups to To implement these rules, it is helpful to also show on
maximize variability between subgroups and to mini- the chart the lines for
2 SE (warning limits) in
mize variability within subgroups. As an example, if addition to the control limits. These rules should be used
we are sampling four bottles from a filling line each with discretion; studies have shown that the use of the
 3502 Statistical Process Control and Process Capability
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Table 2 Averages and ranges calculated for each subgroup for the current data
Subgroup Data Average Range
1 299.7 298.7 300.2 301.3 301.2 300.2 2.6
2 301.7 297.8 299.8 301.1 298.9 299.9 3.9
3 299.3 298.3 298.2 299.0 299.2 298.8 1.2
4 297.9 299.4 299.6 300.1 299.6 299.3 2.3
5 299.7 299.6 301.3 299.9 299.8 300.1 1.7
6 299.5 302.0 300.9 302.4 299.3 301.2 7.0
7 301.7 298.4 300.5 300.9 301.9 300.7 3.5
8 299.9 299.5 300.7 299.6 300.8 300.1 1.3
9 298.6 299.2 298.5 299.6 300.0 299.2 1.5
10 300.0 299.7 302.2 298.3 299.3 299.9 3.9
11 297.4 301.4 298.7 299.3 300.8 299.5 4.0
12 300.5 300.9 300.6 298.6 298.9 301.9 2.2
13 300.7 300.3 299.1 299.8 300.1 302.0 1.6
14 300.6 300.1 299.1 301.9 300.5 302.4 2.8
15 300.1 300.8 300.9 299.4 299.1 302.0 1.8
16 301.1 298.8 298.4 300.7 300.6 301.9 2.7
17 302.2 301.4 301.3 300.1 300.0 303.0 2.2
18 300.5 300.0 298.9 298.2 300.8 301.7 2.6
19 300.4 300.6 300.2 299.0 301.2 302.3 2.3
20 299.7 299.2 299.2 299.6 299.5 301.4 0.5
Sums over subgroups 6017.6 51.6
Averages over subgroups 300.9 2.6

supplemental rules causes an increase of ‘‘false alarm time period. Once per shift (or, more frequently, if
signals’’ vs. using the
3 SE control limits only.[6] necessary), a complete set from all heads (a ‘‘head
check’’) should be taken and these data can be exam-
ined for necessary individual head adjustments.
Control Charts for Multiple Processes
There are two types of special causes in multiple
processes: those that affect a single head (e.g., plugging),
Many processes consist of multiple streams, such as
multiple-head fillers for powder fills or multicavity
molds for tablet compression. Each stream must be Table 3 Average run length to detect a shift of hs vs.
considered a separate process.[7] There are two general subgroup size
situations:
Subgroup size (n)

(1) The stream number can be identified at time of h 1 2 4 6 10 16

product inspection. 0.5 167.3 92.4 44.0 26.4 12.8 6.3
(2) The stream number cannot be identified. 0.6 124.4 63.9 27.8 15.9 7.4 3.6
0.8 72.3 32.4 12.4 6.7 3.1 1.7
For case (1), a group control chart could be main- 1.0 44.0 17.7 6.3 3.4 1.8 1.2
tained on all streams but it suffices to plot only the
1.2 27.8 10.4 3.6 2.1 1.3 1.0
two streams corresponding to the highest and lowest
values. The chart limits are set up for a single stream 1.4 18.3 6.5 2.4 1.5 1.1 1.0
and run rules should not be used. If the output of 1.6 12.4 4.3 1.7 1.2 1.0 1.0
the heads is highly correlated, then a single chart on 1.8 8.7 3.1 1.4 1.1 1.0 1.0
one stream may be used as a surrogate for all streams. 2.0 6.3 2.3 1.2 1.0 1.0 1.0
Case (2) often applies to high-speed fillers with a 2.5 3.2 1.4 1.0 1.0 1.0 1.0
large number of heads. For this situation, a random
3.0 2.0 1.1 1.0 1.0 1.0 1.0
sample of a subset of the heads can be taken at each
Statistical Process Control and Process Capability 3503

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Statistical–
and those that affect all heads (e.g., viscosity). The head technical support people, and plant management.
check addresses the first kind, and the control chart of The team must decide on the most critical variables
random samples addresses the second kind. Head-to- to track, and these variables should be kept to a
head variation can be considered a special cause, but minimum.
one that cannot be eliminated entirely. Ryan[2] discusses three phases of control chart
usage:

Other Control Charts Phase I—retrospective analysis of historical data

for process assessment
A standard deviation chart can be used instead of the Phase II—process monitoring in real time to find
Range or R-Chart, where the sample standard deviation and control special causes
qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
Pn ffi
2 Phase III—process monitoring to confirm that the
s is calculated as s ¼ i¼1 ðX i  XÞ =ðn  1Þ . The
process remains in statistical control.
statistic s is considered to be more ‘‘efficient’’ than the
range R because s makes use of all the data instead of In Phase I, if data are not already available, the first
the two extreme data points in the subgroup. However, step is to set up a plan for gathering and charting the
the R-Chart is easier to use on the shop floor and is data. These charts should be kept in the operating area
recommended for subgroups sizes up to seven, where R and are usually manually plotted. Operators are
is at least 90% as efficient as s. Control chart factors encouraged to record all process adjustments as well
for the s chart can be found in Ref.[3]. as to record anything that might have happened to
The Individual — Moving Range Chart is used the process. These charts may be part of the operating
when the subgroup size is one. If the process deals with records. A minimum of 100 data points is recom-
bulk product in a batch or a continuous process, only mended before control limits are calculated and placed
single measurements may make sense because multiple on the charts.[2] Because most of the subgroups are of
measurements would reflect only the assay and sam- size four to five, this amounts to approximately 20–25
pling variability, not the process variability. Because subgroups. The initial plots may simply be Run
there is no measure of variability in a single measure- Charts, where the averages are plotted. The team
ment, the moving range, or the absolute difference should be discussing potential special causes that might
between two successive individual values, is calculated. affect the process variable being plotted.
The average moving range performs the function that In Phase II, the team should immediately react to an
the average range does in the Shewhart Chart. The out-of-control signal and investigate for a special
control limits for the Individual Chart are computed cause. During this phase, the team should record any
using an A2 value of 2.66. The UCL for the Moving process information that might be relevant in special
Range Chart uses a value of D4 equal to 3.267. cause identification. This phase is complete when the
Two alternatives to the Shewhart control chart, process is in control for an extended period of time
which are more complicated to calculate but generally with infrequent signals (say, one in every 100–300 sub-
more effective to detect small shifts, are the Cumulat- groups). The control limits should be recalculated if a
ive Sum (or Cusum) control chart and the Exponen- process change has occurred. They may also be recal-
tially Weighted Moving Average (EWMA) control culated at some specified frequency, such as every 50
chart. These control charts will not be discussed here, subgroups or annually, depending on how often the
but are described in standard references.[1,2] observations are taken.
The P-Chart is used when the statistic plotted is the During Phase III, the process is monitored for
fraction defective p of n product units. The C-Chart is instances of special causes. When special causes cannot
used when the statistic plotted is the count c of defects be entirely eliminated, an Out-of-Control Action Plan
or occurrences of an event in time or space. These (OCAP) can be developed for routine use by operating
charts are termed ‘‘attribute charts’’ and are also personnel.[2] The OCAP comprises three features: acti-
described in standard references.[1,2] vators, checkpoints, and terminators. The activators
define the conditions that signal when the OCAP must
be activated, and the control chart usually performs
Developing and Maintaining a Statistical this function. The checkpoints instruct the operator
Process Control Program to investigate specific items as possible special causes
for the out-of-control condition. Once a checkpoint
An SPC program can be valuable when used on pro- has been identified as a special cause, a terminator calls
cesses where production is steady. The best chance for for a specific action to be applied to resolve the problem.
successful application occurs when a team is formed, The purpose of SPC is to reduce variability and to
comprising the plant operators, first-line supervisors, improve the quality of process output. Such programs
 3504 Statistical Process Control and Process Capability
Sterilization
Statistical–

derive benefits through reduced costs of manufacture,

better quality, fewer troubleshooting crises, and
improved relationships with customers. In a strict LSL USL
sense, SPC is a tool for getting a process into statistical
control, not for making process adjustments. For mak-
ing process adjustments, while data continue to be Mean
charted, the centerline is usually set at the target value
and process adjustment schemes are needed. Without
these schemes, operators will tend to over adjust. Fig. 3 Comparison of process output vs. specifications for
One reference that discusses such manual process CP ¼ 1.
adjustment schemes in detail is Ref.[4]. Two summary
articles on the same topic can be found in Refs.[8,9].
when the process is centered and in control.
The defects are depicted as the area under the
PROCESS CAPABILITY curve outside the specification limits, and the
process is clearly not capable of producing a
Defining Process Capability Indices product that consistently meets specifications.

As stated earlier, a process capability analysis assesses CP ¼ 1: For this process, the m
3sC range
the process performance relative to product specifica- coincides with the specification limits; therefore
tions. Specification limits are denoted as LSL for the CP ¼ 1 (Fig. 3). Assuming that the process is
lower specification limit, and as USL for the upper speci- centered and in control, the defect rate is
fication limit. In some cases, there is only a single speci- 0.27% if the process output is normally distrib-
fication limit, either a lower limit or an upper limit. uted.
Two process capability indices are in common use, CP > 1: For this process, the m
3sC range is
and they are as follows: much tighter than the specification limits, so that
CP > 1 (Fig. 4). The process has room for slight
(1) The process-centered index CP ¼ (USL 
shift and drift without producing defects.
LSL)/6sC
(2) The process-shifted index C PK ¼ MIN If a process is not centered, than a more meaningful
[(USL  m)/3sC ,(m  LSL)/3sC ] process capability index is CPK. This is shown in Fig. 5.
CPK < 1: For this process, CP > 1, but the process
where m is the process average and sC is the standard is not centered; therefore defects are produced. For this
deviation measuring the inherent process variability. reason, CPK < 1 better reflects this situation (Fig. 5).
The CP index can be used only when there are two- Another definition of CPK that might be easier to
sided specifications. These indices are defined so that understand is CPK ¼ j m  nearest spec limit j /3sC.
higher is better. The relationship between CP and percent defective
Figs. 2–4 illustrate the meaning of the CP index for (assuming a normal distribution) is given in Table 4.
the following situations: CP < 1, CP ¼ 1, and CP > 1. For these indices to be meaningful as descriptors of
The three situations can be described as follows: process capability, the process must be in a near state
of statistical control. A perfect state of statistical con-
CP < 1: The m
3sC range of this process distri- trol is rarely attainable. A criterion for near-statistical
bution (the area under the curve) is wider than control is that out-of-control signals appear about
the specification limits, so that CP < 1 (Fig. 2). once in every 100–300 subgroups. This is usually the
A large percentage of rejects is produced even case in Phase III of the SPC program. Until that point

LSL USL LSL USL

Mean Mean

Fig. 2 Comparison of process output vs. specifications for Fig. 4 Comparison of process output vs. specifications for
CP < 1. CP > 1.
Statistical Process Control and Process Capability 3505

Sterilization
Statistical–
Table 5 Relationship between sP/sC
LSL USL and the percentage of total variation
represented by special cause variation
rP/rC r2S/r2P (%)
1.30 41
Mean 1.25 36
1.20 31
Fig. 5 Comparison of process output vs. specifications for 1.15 24
CPK < 1. 1.10 17
1.05 9
is reached, the process capability indices should reflect 1.00 0
only the process potential should the special cause varia-
tion be eliminated. Some of the many cautions asso-
ciated with the use of capability indices are described Estimating Process Capability Indices
throughout this entry and detailed in Refs.[10,11].
To estimate process capability indices from data, the
estimates replace the true process values in the follow-
Defining Process Performance Indices ing formulas as follows:
The process-centered index
Sometimes an analysis of process capability is desired
when the process is not in a state of strict statistical ^ P ¼ ðUSL  LSLÞ=6^
C sC
control. When this occurs, process performance indices
should be used. Process performance indices can be
The process-shifted index
defined as: PP ¼ (USL  LSL)/6sP, and PPK ¼
MIN[(USL  m)/3sP,(m  LSL)/3sP], where sP is
^ PK ¼ MIN½ðUSL  XÞ=3^
C sC ; ðX  LSLÞ=3^
sC 
now the overall standard deviation of the process out-
put characteristic.
For a process that is not in a state of strict statistical where X is the estimated process average and s ^C is the
control, sP is greater than sC, the inherent variation, estimated standard deviation measuring the inherent
because there is additional variation because of process process variability. The estimates of X and s ^C can
shift and drift. This additional variation component is come from a prior control chart analysis, where
sS, where the ‘‘S’’ in sS denotes special cause variation. ^C ¼ R=d2 .
s
Because variances, not standard deviations, are Use of the C^P estimate assumes that the process is
additive, these components can be represented as centered at or near the exact middle of the specification
s2P ¼ s2C þ s2S. Another way to state the objective range of a two-sided specification. This represents the
of SPC is to reduce sS as close to zero as possible. best capability achievable. The C^PK estimate assesses
One criterion used to assess whether a process is the process with no assumption that the process is cen-
near statistical control is the closeness of the ratio tered, and this estimate is always equal to or less than
sP/sC to 1. Table 5 shows the relationship between this the corresponding C^P estimate. If the distribution of
ratio and the percentage of total variation represented the output is close to a normal distribution and the
by special cause variation. capability index is based on a large and representative
For example, when sP/sC ¼ 1.2, the percentage of sample size, then acceptance values can be established.
special cause variation is 31%, which may represent a Some industries define a process with C^PK ¼ 1 as
practical goal for attaining ‘‘a near state of statistical barely capable and one with C^PK ¼ 1.33 or higher
control.’’ as capable. (Note that a C^PK ¼ 1 represents a situa-
tion where the process is centered and the specification
range is equal to 6^sC .)
Table 4 Relationship between CP and percent defective
CP %Def CP %Def Estimating Process Performance Indices
0.6 7.19 1.0 0.27
0.7 3.57 1.1 0.097 The estimates of the performance indices are defined
analogously to the capability indices as P ^P ¼
0.8 1.64 1.2 0.032
ðUSL  LSLÞ=6^ sP and P ^ PK ¼ MIN½ðUSL  XÞ=
0.9 0.69 1.3 0.001
sP ; ðX  LSLÞ=3^
3^ sP , using the estimate s
^P , which is
 3506 Statistical Process Control and Process Capability
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Statistical–

the estimated overall standard deviation of the process Table 6 The 95% two-sided confidence intervals for two
output characteristic. Thus s ^P , the estimate of sP, different sample values of CˆP for different sample sizes
includes not only the estimated inherent deviation s ^C , C^P ¼ 1.00 C^P ¼ 1.33
but also the estimated variation due to process shift
and drift from special causes. Therefore s ^P > s ^C for n Lower Upper Lower Upper
a process not in statistical control. 10 0.55 1.45 0.73 1.93
Process performance indices strictly apply only to 30 0.74 1.26 0.99 1.67
the data set being examined because the process is 100 0.86 1.14 1.14 1.51
not predictable into the future when it is not in statisti-
200 0.90 1.10 1.20 1.46
cal control. However, in practice, the indices are often
interpreted to provide an assessment of the process as
it currently exists. where w21  a/2,n1 and w2a/2,n  1 are the lower and
If the process is not in a state of strict statistical con- upper a/2 percentage points of the chi-square distri-
trol (i.e., stable and predictable), every attempt should bution with n  1 degrees of freedom.
be made, when practical, to improve control and to put This interval is constructed to contain the true
the process into as close to a state of statistical control unknown index CP with a specified degree of confi-
as possible. If the process cannot be put into statistical dence 100(1  a)%. For example, 95% two-sided con-
control, enough time points should be collected to be fidence intervals for two different sample values of C^P
able to assess the full degree of shifts and drifts of for different sample sizes are as shown in Table 6.
the process and to determine whether the data are As can be seen from the table, if a sample C^P of 1.00
indicative of the long-term process with all of its shifts is obtained from a sample of 30 values, then with 95%
and drifts. As Pruett and Schneider[12] indicate, PPK is confidence, it can be stated that the true process capa-
a measure of process capability over a period of time. bility CP is between 0.74 and 1.26. If, instead, one is
If extensive data can be obtained, the ability of the pro- only interested in one direction (e.g., how low the true
cess to meet its specifications can be evaluated, with CP index might be based on a sample of size n), a one-
care, by the estimate P^PK. If extensive data cannot be sided lower confidence interval could be generated.
obtained, P^PK can provide only the capability of that If, on the other hand, one is interested in determin-
particular set of data to meet the specifications. ing the minimum acceptable sample process capability
Before the special causes are fully identified and
index C^P required to have some stated confidence
controlled, P^PK estimates the process performance that
100(1  a)% that the true CP is above some value
the customer currently experiences, and C^PK estimates
(e.g., CP ¼ 1.0), one can use the following equation
the potential process capability attainable when the
for a particular CP, sample size n, and risk level a:
process is brought more closely into a state of statisti- qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
cal control. As the process approaches a state of sta- the minimum C^P is CP = w2 1a;n1 =n  1. This can
tistical control, P^PK approaches C^PK. Similarly, the be useful for determining sample sizes during protocol
estimates P^P and C^P are used when the process is generation where a particular minimum CP is required
centered within two-sided specification limits, or to for protocol acceptance. For example, the sample
reflect what the capability would be if the process were capability indices that would be needed to have at least
centered. a 95% confidence that the true process capability is at
least 1.00 (or 1.33) based on various sample sizes are
shown in Table 7.
Precision of Estimates of Capability and Confidence intervals for any of the other indices of
Performance Indices interest (PP, CPK, or PPK) have sometimes been esti-
mated in practice by using the above equations but
It is important to remember that the capability and by substituting the index of interest. However, these
performance indices calculated from the data are only
estimates and thus have uncertainty associated with Table 7 Sample capability indices that provide at least a
them. Confidence intervals can be calculated for these 95% confidence that the true process capability Is at least
indices to reflect this imprecision.[1] For example, a 1.00 or 1.33 based on various sample sizes
100(1  a)% two-sided confidence interval about the n CP ¼ 1.00 CP ¼ 1.33
estimated process capability index C^P is:
10 1.65 2.19
sffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi sffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi 30 1.28 1.70
w21a=2;n1 w2a=2;n1 100 1.13 1.51
^P
C ^P
 CP  C
n  1 n  1 200 1.09 1.45
Statistical Process Control and Process Capability 3507

Sterilization
Statistical–
intervals should only be considered as approximate. for the known non-normal distribution (to obtain
More statistically based approximations for these 95% confidence). One common distribution that is
intervals are discussed in Refs.[13,14]. One-often used effective in fitting a broad range of distributional
approximate confidence interval for CPK assumes that forms is the gamma distribution.
the distribution of C^PK is normal and a Taylor series  Use the index ‘‘as is’’ as a relative measure and note
approximation is used to construct a 100(1
a)% in report to this effect. Even if the data are non-
two-sided confidence interval for CPK as follows: normal but symmetrical, the calculations are gener-
sffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi ally reasonable. However, if the data distribution is
^2 heavily skewed, one should be hesitant to use this
^ PK  Za=2 1 þ C PK
C particular option.
9n 2n
2
 Omit the computation of capability and perform-
where Za/2 is the upper a/2 quantile of the standard ance indices because the values so obtained would
normal distribution. For example, to generate a 95% be misleading.
two-sided confidence interval, use Za/2 ¼ 1.96.
As can be seen from the above examples, reported
Process Capability Example
estimates of capability and performance indices based
on small sample sizes are potentially misleading and
This example studied the process capability of capsule
should not be used unless relatively large samples are
weight for a hypothetical prescription drug. The speci-
obtained, or they are accompanied by a confidence
fication limits were 290–350 mg. Four capsules were
interval when reporting the results.
sampled from the process every 20 min and weighed.
There were 25 subgroups of four data points each for
a total of 100 data points. The results are summarized
Non-normal Distributions
in Fig. 6.
The control chart analysis indicated that the process
The use of the 6s factor in defining the process capa-
was in a state of statistical control because none of the
bility indices assumes that the data follow a normal
averages and ranges was outside its respective control
distribution. It is important to remember that large
limits. The data distribution (histogram) was deemed
values of these capability indices (indicating a very cap-
reasonably close to a normal distribution (the bell-
able process) may be derived from incapable processes
shaped curve superimposed on the histogram) because
when the data are non-normal. Similarly, data from a
the cumulative data distribution was close to a straight
quite capable process may result in low capability
line on the normal probability plot.
index values just because the data are non-normal.
^P ¼ 6.89
In this study, the variability estimates were s
Normality of the data can be checked in a variety of
mg and s ^C ¼ 6.60 mg, both in close agreement because
ways. Routine plotting of histograms is a good first
the process was in statistical control. The C^PK estimate
step in assessing whether the data are normal. Some
was 1.32, which was lower than the C^P estimate of 1.52,
of the more detailed techniques for determining
because the process was not exactly centered between
whether a set of data is normal are: probability plot-
the specification limits. The process performance esti-
ting, the chi-square goodness of fit test, and the
mates did not apply because the process was in statistical
Anderson–Darling statistic.[15] If it is determined that
control, but were calculated by the package and were
the data are not normal, four options are available:[16]
close to the capability estimates. (Note: In the Minitab
output, the estimates do not have the caret (^) over the
 Apply a transformation to the data to make the
symbol, but they are in fact estimates.)
transformed data normal. If the distribution is
If the process were not in a state of statistical
skewed to the right, one might try a log, inverse,
control (stable and predictable), then the P^P and P^PK
square root, or cube root transformation of the
estimates could be used to judge the process perform-
data to make the data normal. If the data are
ance at the time the data were collected. The C^P and
skewed to the left, an exponential, squared, or
C^PK indices could be used to assess the process poten-
cubed transformation might be applied. A histo-
tial if the process were brought into a state of statistical
gram can be applied before and after the transforma-
control.
tion to assess the ability of the transformation to
make the data normal. It is important to remember
that the transformation must be applied to the USL Relationship between Statistical Control and
and LSL, in addition to the data, before computing Process Capability
the capability index of interest.
 Assume a known distribution and use the multi- To summarize, it is important to remember that a pro-
pliers that correspond to the 2.5% and 97.5% points cess may be in statistical control but still not capable.
 3508 Statistical Process Control and Process Capability
Sterilization
Statistical–

X bar and R Chart Capability Histogram

329
UCL=326.0
321

Mean
Mean=316.1
313
LCL=306.2
305
300 315 330
Subgr 0 5 10 15 20 25
Normal Prob Plot
30 UCL=30.98
Range

20 _
R=13.58
10
0 UCL=0
300 315 330
Last 25 Subgroups Capability Plot
Within
330 StDev: 6.59538 Process Tolerence
Cp: 1.52 Within
Values

320 Cpk: 1.32 Overall

310 Overall
StDev: 6.88567 Specifications
300 Cp: 1.45 290 350
Ppk: 1.26
0 5 10 15 20 25
Subgroup Number

Fig. 6 Summary of capsule weight data using the statistical package, MinitabÕ. (From Minitab Inc., State College, PA. For
details, refer to www.minitab.com.)

In a strict sense, it is not appropriate to talk about  Powder characteristics (mean particle size, bulk
whether a process is capable if it is not in a state of density, etc.)
strict statistical control (because it is not stable and  Microbial counts
predictive of the future); in practice, the indices are  Drug content application (nasal spray, etc.)
often used to provide an assessment of the process as  Fill weight and fill volume
it currently exists. Caution has been given to the reader  Air/oxygen-purged systems
to make sure that before an assessment is made of a  Leachables, residuals
process that is not in statistical control, enough data  Liquid characteristics (viscosity, refractive index,
have been collected to cover the full range of shifts etc.)
and drifts in the process. With this information in  Consumer complaints, industrial safety measure-
hand, it is possible to claim, with some caution, that ments, etc.
a process is capable of staying within specifications
although it is not in statistical control. However, even Bringing processes into or near a state of statistical
though the process may be capable, there are still many control will improve processes by making them less
benefits in working toward the elimination of all variable, centered closer to target, and allow the manu-
special cause variation. facturer to make a product that will more consistently
meet product specifications. This benefits both the
manufacturer and the consumers who use their pro-
APPLICATION TO PHARMACEUTICALS ducts. The use of SPC methods to evaluate and to
improve processes not only can be applied to product
Typical Applications characteristics such as tablet weight and tablet hard-
ness, but also to product performance measures such
The concepts discussed in this paper are appropriate as consumer complaints, line down time, and industrial
for many applications in pharmaceuticals. Some of safety measurements. An SPC approach to process
these areas are as follows: improvement can also lead to reductions in fill
overages, reductions in waste, as well as reductions in
 Drug potency batch failures. By eliminating special cause variability,
 Other finished product attributes (e.g., content it becomes easier to monitor a process to ensure that new
uniformity, dissolution) special causes do not find their way into the process.
 Tablet/capsule in-process characteristics (weight, As mentioned earlier, as part of a process capability
thickness, hardness, disintegration, etc.) study, an assessment of variability is conducted.
Statistical Process Control and Process Capability 3509

Sterilization
Statistical–
Variability may include both process and measurement For example, some process monitoring schemes use
variability. This may be especially important when an average of 10 tablets to track tablet weight. The 10
measuring such attributes as potency, content uniform- tablets are sampled and measured in a single weighing
ity, and dissolution. It is important to be able to separ- operation at each sampling time, and the average
ate process from measurement variability during any weight is computed and plotted. In this situation, the
investigation of how to improve processes. The sam- state of statistical control of tablet weight can be
ples may need to be tested in a designed fashion to assessed using Individual Moving Range Charts on
be able to correctly separate these sources of vari- the tablet averages, but such data may not be appli-
ability. Measurement variability can be independently cable for process capability studies when the specifi-
addressed through assay validation studies. cation is based on individual tablets.
It is important to remember that a process may be
in statistical control but still not capable. Range stud-
Analysis of In-Process Data ies for the critical parameters are an effective way to
determine whether the specification limits are tighter
Control charts are an excellent analysis tool to both than they need to be, or, if no specifications exist, to
monitor and improve in-process performance during aid in their development. A range study is intended
process development and later during production, to determine the high and low ranges for which the
where it is desired to follow process characteristics over parameter is able to demonstrate an ability to meet
time within batches or runs. The most common exam- end-product requirements; for example, high and low
ples of tablet process characteristics that are measured hardness ranges that meet both dissolution as well as
in-process are weight, thickness, and hardness. The physicals (e.g., no chipping) help to define appropriate
parameters measured need to be controllable so that in-process specification limits. If the process is not cap-
adjustments can be made. During the initial runs, it able, this may be because the specifications had orig-
is desirable to limit process adjustments to a minimum inally been set at
3s limits and not what defines
to observe the process in its natural state. Any adjust- acceptable product, as is the purpose of a range study.
ments made should be recorded and explained. Out-of- If this is what was done, capability indices of around
limit results should never be removed prior to perform- 1.0 must be expected.
ing a process capability analysis. If special cause varia-
tion is detected, then process improvements should be
made to eliminate the special cause variation. Analysis of End-Product Data (e.g., for
For new products, capability should be evaluated Validation, Annual Product Review, etc.)
and improvements made to the process during devel-
opment and scaleup, before validation is performed. Capability and performance indices are appealing as a
It should be conducted on the actual full-scale equip- management process assessment tool because they tie
ment that will be used during routine production. together the process performance (as measured by
Our experience has been that when evaluating the sample mean and sample standard deviation) with
in-process tablet and capsule parameters during the product specifications into a single measure. Pro-
production, a good setup is important to ensure that cess performance indices are sometimes used as accept-
the process starts out at the target value. Addition- ance criteria during validation, as well as Annual
ally, the practice of 1) shutting down the line for Product Review; however, they need to be used very
breaks and lunch and 2) allowing hopper runout carefully. Sample sizes may be too small to provide
during processing can cause added variability because reliable estimates, especially if these indices are used
of segregation of the powder. to determine the pass/fail of a validation study or an
Product flow characteristics and interdependencies Annual Product Review. Emphasizing the use of capa-
between parameters, such as tablet weight, thickness, bility indices without the efforts required to initially
and hardness, may prevent the parameters from each put the processes into statistical control misses the
being in strict statistical control. Special studies testing point. Although not recommended by the authors as
weight, thickness, and hardness on the same tablets will strict acceptance criteria for validation or Annual Pro-
aid in better understanding this interrelationship duct Review, capability indices, if used carefully, do
between these product characteristics. For such char- provide a useful yardstick for comparison of perform-
acteristics, the in-process specifications may be on ance over time periods, or for comparison between
either the individual values or the averages (e.g., the products for helping in determining priorities for pro-
average of 10 hardness values), or both. When the cess improvement efforts. These indices also provide a
average is being evaluated, process capability indices yardstick to measure the effect of any improvements
may not be appropriate for evaluating the ability to made to the process in a ‘language’ that can be easily
meet specifications based on individuals. understood. There are also times when there are just
 3510 Statistical Process Control and Process Capability
Sterilization
Statistical–

better ways to define process capability for solid dos- has occurred. In addition, once stability changes over
age forms, possibly by tying it to the end-product test time, batch processing differences, slight biases in the
requirements [e.g., the ability to meet the multiple- analytical method, and typical measurement and pro-
stage United States Pharmacopeia (USP) tests for cess variability are accounted for, defective levels
content uniformity and dissolution]. may not always be in the parts-per-million range as
The use of common pharmaceutical ingredients and is required for a CPK of 1.33. In fact, if measurement
the tendency to ‘‘gang test’’ assays in the laboratory error is large, arbitrary CPK goals may actually be
may prevent many of our end-product attributes from impossible to meet no matter how much the process
being in strict statistical control when measured over is improved.[11]
time. Therefore it is not recommended that a vali- Because, in the pharmaceutical industry, being the
dation study or Annual Product Review require that first to market a new drug is of utmost importance,
a process be in statistical control to be considered taking the time to put processes into statistical control
acceptable, not even for in-process parameters such is too often perceived by companies as slowing down
as tablet weight, thickness, and hardness. Processes the process of product introduction and not as added
that are not in a state of strict statistical control can value. This attitude needs to change. Companies may
be capable of consistently meeting specifications and also feel that their reward for improving process con-
can be validated. However, if processes are not in sta- trol and reducing variability is that the FDA will
tistical control, efforts should always be made to tighten specifications further, thereby risking the pro-
eliminate special causes and get them into as near a duction of out-of-specification product. A company’s
state of statistical control as possible. The validation interest in improving processes is often discouraged
and Annual Product Review data can even be helpful by a need to revalidate or refile with the FDA when-
in determining how processes can be improved. ever significant changes are made to the filed NDA.
Although these comments have been applied to end- The authors feel that there needs to be discussions
product data, many of these same comments also apply between industry and the FDA on these issues so that
to in-process data as well. industry is rewarded for the improvement of their pro-
cesses and everyone, most importantly the consumer,
wins. This need for cooperation between the FDA
and industry to move the pharmaceutical industry
Limitations Within Pharmaceuticals toward real improvements was voiced by Woodcock
of the FDA in Ref.[18].
As discussed earlier, processes in the pharmaceutical
industry may be difficult to always get into a state of
strict statistical control. Interdependencies may exist
between attributes such as the weight, thickness, and CONCLUSIONS
hardness of in-process tablets because of product flow
characteristics within the batch. Data from sequen- The use of SPC to help improve processes and the use
tially generated batches may be dependent because of of process capability to evaluate the ability of processes
the use of common raw materials, or being tested on to routinely meet specifications are described in detail.
the same day. But the benefits derived from using the They are companion tools that should be used in con-
techniques discussed in this entry to improve existing cert. These tools are described in sufficient detail to
processes and to keep them under control are many. allow interested practitioners to correctly apply them
The effort is worth the hard work required to do it. within their workplace. Examples of their use are pro-
Nash[17] points out that what might be possible in vided for reinforcement as are the assumptions being
other industries may not always be possible for made when using them. Some of the limitations asso-
pharmaceutical processes in that a CPK of 1.33 may ciated with their use are also discussed, especially in
not always be achievable. Specifications should be the context of application within the pharmaceutical
customer-directed limits that ensure safety and efficacy, industry.
not limits to control the consistency of process. Inter- With the writing of this entry, the authors hope to
nal control limits should be used for this purpose. encourage the proper use of these important process
Many of our specifications are either provided to us improvement and process evaluation tools, especially
by the USP or influenced by the Food and Drug within the pharmaceutical industry. We also hope that
Administration (FDA), where industry is encouraged some of the barriers to their implementation within the
to set specifications as tight as the process will allow. pharmaceutical industry that were discussed within
Companies are sometimes asked to agree as to this entry can be overcome through cooperation with
what they view as tight specifications before the full the FDA and a mutual desire to improve pharmaceu-
experience that comes with additional production tical processes.
Statistical Process Control and Process Capability 3511

Sterilization
Statistical–
REFERENCES 10. Kane, K. Process capability indices. J. Qual. Technol. 1986,
18 (1), 41–52.
1. Montgomery, D.C. Introduction to Statistical Quality Con- 11. Gunter, B. The use and abuse of Cpk, Part 4. Qual. Prog.
trol, 4th Ed.; Wiley, 2001. 1989, 22 (7), 86–87.
2. Ryan, T.P. Statistical Methods for Quality Improvement, 12. Pruett, J.M.; Schneider, H. Essentials of SPC in the Process
2th Ed.; Wiley, 2000. Industries, 2nd Ed.; Instrument Society of America,
3. Manual on Presentation of Data and Control Chart Analy- 1996.
sis, 7th Ed.; ASTM International, 2002. 13. Rodriguez, R. Recent developments in process capability
4. Box, G.E.P.; Luceño, A. Statistical Control in Monitoring analysis. J. Qual. Technol. 1992, 24 (3), 176–187.
and Feedback Adjustment; Wiley, 1997. 14. Kushler, R.H.; Hurley, P. Confidence bounds for
5. Statistical Quality Control Handbook; Western Electric capability indices. J. Qual. Technol. 1992, 24 (3),
Company, 1956. 188–195.
6. Champ, C.W.; Woodall, W.H. Exact results for Shewhart 15. Shapiro, S. How to Test Normality and Other Distributional
control charts with supplementary runs rules. Techno- Assumptions; How To Series; ASQ, 1990; Vol. 3.
metrics 1987, 29 (4), 393–399. 16. Quality Assurance for the Chemical and Process Indus-
7. Nelson, L.S. Control chart for multiple stream processes. J. tries, a Manual of Good Practices, 2nd Ed.; ASQ Quality
Qual. Technol. 1986, 18 (4), 255–256. Press, 1999; 42.
8. Box, G.E. George’s column. Qual. Eng. 1991–1992, 4 (1), 17. Nash, R.A. Understanding the process capability index
143–151. concept. J. Valid. Technol. 1998, 4 (3), 202–204.
9. Box, G.E. George’s column. Qual. Eng. 1991–1992, 4 (2), 18. Regulatory Leeway sought for process analytical tech-
331–338. nology. Gold SheetFebruary 2002, 36 (2), 1–19.
Sterilization
Statistical–

Sterilization: Dry Heat

Andrea Chieppo
Thomas Kupiec
Analytical Research Laboratories, Oklahoma City, Oklahoma, U.S.A.

INTRODUCTION DRY-HEAT STERILIZATION KINETICS

Sterilization means the destruction of all life. The aim of Sterilization is a probability function. In the pharmaceu-
sterilization is to destroy the ability of microorganisms to tical industry, an item is deemed sterile if there is less than
survive and multiply with the oldest and most recognized 1 chance in 1,000,000 that viable microorganisms are
agent of destruction being heat.[1] Dry heat is the method present in the sterilized article or dosage form. Therefore
of choice for sterilizing heat-stable items that might not there is a 106 probability of non-sterility.[6] In all sterili-
be adequately penetrated by steam or are damaged by zation processes, the inactivation of microorganisms
moisture.[2] Dry-heat sterilization is frequently used in develops as a first-order chemicalreaction, i.e., at a rate
the pharmaceutical industry; items often sterilized by which is approximately logarithmic.[7]
dry heat include powders, oils,[3] petrolatum, glassware, The following equation typically describes the order
and stainless-steel equipment.[2] of microbial death:
Dry-heat sterilization is generally a less complicated
process than steam sterilization; it is, however, rela- K ¼ 1=tðlog No  log N Þ
tively slow and requires higher temperatures and/or
longer exposure times. This is because of the fact that where K ¼ a constant, assuming logarithms to the base
microbial lethality is lower with dry heat than that for 10 and depending on the organism, temperature, and
steam at the same temperature.[2] There are various substrate, t ¼ time of exposure in min, No ¼ initial
temperatures and periods of treatment for dry heat number of viable organisms, and N ¼ number of viable
depending on the pharmacopeia. The U.S. Pharmaco- organisms at the end of the time interval.[7]
peia (USP) states that the dry-heat sterilization process Using natural logarithms, the exponential form of
for containers for sterile pharmaceutical products the equation is:
should be at a temperature of 160–170
C for a period
of 2–4 hr. The British Pharmacopeia states that items N ¼ No eKt
sterilized by dry heat should be kept at a temperature
not less than 160
C for at least 1 hr. For the Pharma- It was found that a 90% reduction in the microbial
copeia Nordica, the recommendation is 30 min at population resulted in the following equation for K:
180
C. Different materials and sterilization equipment
used account for the discrepancies between these K ¼ 1=tðlog No  log 0:1No Þ
pharmacopeias, but there is also a lack of sufficient
information concerning dry-heat sterilization.[4]
Dry-heat processes have two main targets: K ¼ 1=tð1Þ ¼ 1=t
microorganisms and their by-products, pyrogens or
endotoxins. Depyrogenation is the process that t ¼ 1=K
destroys the chemical activity of these by-products.
Destruction of microorganisms and endotoxins by Time t is defined as the decimal reduction time or D
dry heat is considered an oxidative process, which is value. Therefore
almost a combustion.[5] Depyrogenation requires a
higher temperature than sterilization and can be sum- D ¼ 1=K or D ¼ 1=10
marized as follows:
Fig. 1 shows a simplified way to determine the D
If an effective dry heat depyrogenation is performed,
value where the amount of time can be determined to
sterilization generally is achieved as well. Effective reduce the microbial population by 90%.[7]
dry heat sterilization can be performed even without The kinetics of dry-heat treatments is comparable to
achieving depyrogenation. If moist heat sterilization that of moist heat sterilization. The organisms that are
is performed, in normal operating conditions depyro- considered to be representatives for dry-heat sterilization
genation is not achieved.[5] processes are spores of Bacillus subtilis var. niger.[4]
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-120012009
3512 Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved.
Sterilization: Dry Heat 3513

Sterilization
Statistical–
Convection Heating Processes

In the pharmaceutical industry, large-scale industrial

ovens are used. These large-scale processes usually
operate to sterilize only because higher temperatures
are required for depyrogenation and this would be
prohibitively long.[6] Convection heating occurs when
heat is transferred through a medium by motion of
its parts. The two types of convection heating include
natural and forced convection heating. Natural con-
vection heating happens as a result of the buoyancy
forces generated by differences in density. In forced
convection heating, high-efficiency particulate air
(HEPA)-filtered air is heated by passage over the elec-
tric heating elements of the oven and then this heat
is transferred from the air to the product by forced
convection.[2] The basic equation for convective heat
transfer is:
Fig. 1 Microbial death rate curves that illustrate concept of
decimal reduction (D values) and probability of survivors. q=c ¼ hc ADT
(From Ref.[6].)
where q/c ¼ the rate of heat transfer by convection
(BTU/hr), hc ¼ the average unit thermal convective
conductance (BTU/hr ft2
F), A ¼ area (ft2), and
A D170 value, which is the time needed to give a 90% DT ¼ the difference between the surface temperature
reduction of the spore population at 170
C, of at least and the temperature of the fluid at some specified
1.5 min may be assumed for these spores.[6] location (
F).[2]
Fig. 2 shows a schematic diagram of a modern
FH Concept forced-convection batch sterilizer. The unit is a two-
door sterilizer where the unloading door leads to the
For dry-heat temperatures other than 170
C, FH values sterile area. The pressure inside the chamber must
are used. The FH concept is comparable to the FO con- continuously be controlled so that the inside is always
cept for moist heat sterilization and references lethality slightly higher than the pressure in the non-sterile
to equivalent times at 170
C. FH values are shown in loading area and slightly lower than the pressure in
units of minutes or seconds, and the calculations of the sterile unloading area.[5]
FH use the same equations as the calculations of FO. There are four stages involved in oven sterilization.
The only difference is that a z value of 20
C is substi- They are 1) drying; 2) heat-up; 3) exposure; and 4)
tuted for 10
C.[6] cooldown. During the drying stage, moisture is driven
off the product to the atmosphere until the air tem-
perature in the sterilizer is around 80
C. At this point,
a baffle closes which allows the temperature within the
DRY-HEAT STERILIZATION PROCESSES
oven to reach the operating level; this is the heat-up
phase. Exposure starts the moment the sterilization
Great strides have been made in developing and
thermal sensor reaches the set temperature. After the
improving other methods of sterilization, but little
timed exposure period, the heating elements switch
has been accomplished where dry-heat sterilization is
off and cooldown begins. Fig. 3 shows a normal batch
concerned. Sterilizers used today are still basically the
process.[6]
same as they were a decade ago.[1]
The two types of processes currently used in dry-
heat sterilization include: 1) dry-heat batch sterilization/ Dry-Heat Tunnel Sterilization
oven sterilization and 2) dry-heat tunnel sterilization.
Process 1 is the type of dry heat unit widely used in the Dry-heat tunnel sterilization is a continuous conduc-
pharmaceutical industry; it uses the principle of con- tion process in contrast to the batch processes in ovens.
vective heat transfer to heat the load. Process 2 is only A continuous conduction process is one in which a pre-
found in large-scale processes, and the main application determined quantity of items is subject to a continuous
of this process is in the sterilization and depyrogenation conduction cycle at a predetermined rate to effect ster-
of glass.[6] ilization and/or depyrogenation.[2]
 3514 Sterilization: Dry Heat
Sterilization
Statistical–

varies directly with the free electron concentration. The

solids with the highest amount of free electrons are pure
metallic solids, and those solids with the lowest con-
centration are the non-metals; therefore pure metals
are the best conductors of heat.[2]
Dry-heat tunnels differ from ovens because energy
is not lost to heating and reheating between cycles as
occurs with ovens. For a majority of the time spent
in the tunnel, the product is subjected to rising tem-
peratures, unlike ovens where the product is held at a
constantly maintained exposure temperature.[6] Tun-
nels basically consist of a thermally insulated ‘‘tunnel’’
which directly connects an upstream cleaning machine
to the downstream sterile area. Inside the tunnel is
a horizontally rotating transport belt made of a
stainless-steel mesh. When a product is sterilized in a
tunnel, it is dried, heat-treated by radiant heat or hot
air, and then cooled. A typical tunnel sterilization
exposure time is 3 or 4 min at 300
C or more. Whether
radiant heat or hot air is used, the internal part of the
tunnel must remain pressurized by ventilation at an
intermediate pressure level that is between the sterile
downstream system and the loading room.[5] Fig. 4
shows a comparison of the two main types of steriliz-
ing tunnels.

Radiant-heat sterilization tunnels

In the pharmaceutical industry, radiant-heat tunnels

have had extensive usage. During this process, energy
flows from a high-temperature body to a lower-
temperature body and the heat is emitted in the form
of finite batches of energy without the aid of an
Fig. 2 Dry-heat batch sterilizer. (From Ref.[4].)

Conduction heating occurs via two different mecha-

nisms. The first mechanism is accomplished by molecu-
lar interaction where molecules at a higher energy level
impart energy to molecules at a lower energy level.[2]
The second mechanism takes place through ‘‘free’’
electrons. The ability of different solids to conduct heat

Fig. 3 Example of a batch process. (From Ref.[2].) Fig. 4 Sterilizing tunnels. (From Ref.[5].)
Sterilization: Dry Heat 3515

Sterilization
Statistical–
intervening medium.[2] Infrared tunnels transmit heat an excessive escape of air from the sterile area to the
by exposing the surface of items to radiation by direct tunnel resulting in the disturbance of the laminar air-
rays. This is accomplished by infrared heaters, located flow and temperature inside the tunnel.[5]
in the roof of the tunnel; when products pass through
the tunnel, their surfaces are heated along with the
internal surface of the tunnel itself. This heat is then
diffused throughout the product by radiation, conduc- EFFECTS OF DRY HEAT ON
tion, and turbulent airflow.[6] MICROORGANISMS AND ENDOTOXINS
The basic transfer equation for radiant heating pro-
cesses is as follows: Inactivation of Microbial Populations

qr ¼ sAT14 As mentioned earlier, inactivation of microorganisms

occurs by oxidation, but other possibilities must
where qr ¼ rate of heat flow in BTU/hr, A ¼ surface also be considered. One possibility is the effect on
area of the emitting object in ft2, T1 ¼ surface tem- DNA. In B. subtilis spores, sublethal temperatures
perature of the emitting object in degrees Rankine, have been shown to induce mutants as a result of
and s ¼ Stefan–Boltzmann constant 0.174  108 depurination. There are also claims that genetically
BTU/hr ft2
R.[2] determined differences in water content could result in
There are disadvantages of IR tunnels; they are the dry-heat sensitivity of spores.[8] A great amount
large, their belt speeds are slow, product heat-up may of interest has been generated relating the water
be less uniform, and they may create particulate prob- activities of spores with their thermal resistance.
lems. These particles are generated from the deterio- The water content of dry microbial cells is important
ration of the heating element and the moving belt. in regards to their destruction rate, and water content
This problem can be helped by installing HEPA filters is determined by the relative humidity of the atmo-
at the entrance of the tunnel and at the cooling zone sphere surrounding the cells; therefore the destruction
junction.[6] rate varies with the relative humidity of the system. In
dry-heat sterilization, water is not present in the liquid
Hot-air laminar flow sterilization tunnels state. If the relative humidity is 100%, then some water
is present in the liquid state. Therefore the relative
The most effective means for the sterilization of air is humidity of a dry-heat sterilization system can be any
incineration. In laminar flow (LF) tunnels, heating value between 0% and 100%. To establish the relative
occurs by the circulation of hot filtered air, which is sterilization effect in dry-heat sterilization, the relative
forced onto the product. The air is then withdrawn humidity conditions must be known in addition to the
by a circulation fan and leaves the product through temperature.[1] At equilibrium, the water activity (Aw)
heating elements below the transport belt and then is inside the cell is equal to the relative humidity of the
brought back into the tunnel through HEPA filters.[5] atmosphere surrounding the microbial cell.[8] Water
This sweeping action of the laminar airflow helps con- activity is used to describe the relative water availability
tribute to the removal of particulate contamination. inside a microbial cell or spore and is related to the
The air speed within the LF tunnel remains around concentration and is expressed as a fraction of the pure
0.5 m/sec. Laminar flow tunnels also have the advan- component in the standard state where:
tage of quicker heating resulting in shorter process
time. This fast heat transfer also contributes to a lower Aw ¼ 1
risk of product contamination from particles.[6]
From the point of view of handling the product, When reporting on dry-heat test data, relative
continuous tunnels are more favorable than batch humidity is measured and controlled instead of water
ovens. The main reason for this is that there is no batch activity. This is because of the fact that biological
work required after the unpacking of the components materials more closely parallel vapor pressure than
and loading of them into the tunnel until the packaged water content, and vapor pressure of the gas atmo-
product is removed from the line after filling. This is sphere surrounding dry microbial cells is more easily
important in large-scale productions, but for easier measured and controlled than the water content inside
isolation of the sterile area, batch ovens are better. This the cell.[1]
is because of the fact that there must be a continuous There are three primary and three secondary vari-
flow of air through the open connection between the ables involved with the mode of action of dry heat.
sterile area and the tunnel. The pressure of the sterile Temperature, water content, and time are the primary
area must always remain higher than that of the variables, and the secondary variables are open and
tunnel. A large difference between the two would cause closed systems, physical and chemical properties of
 3516 Sterilization: Dry Heat
Sterilization
Statistical–

the microorganism and adjacent support, and the gas as the total volume of the enclosure. Changing either
atmosphere.[8] of these parameters will alter the relative humidity,
which then alters the water content in the spore during
Effect of temperature heating.[8]

The most important variable in dry-heat sterilization is

Dry-Heat Destruction of Microorganisms
temperature. Temperature, which is the measure of the
Associated with Soil
heat energy level, is a function of time. One measure
of the change in destruction rate with temperature is
There are two groups of microorganisms that accumu-
the z value. The z values for dry-heat inactivation of
late on an object when it comes in contact with people
microbial spores have been reported from about
and air. The first group includes naked or unprotected
15
C to 30
C.[8]
microbial cells from human or animal origin. The
second group is made up of spores found in particles
Effect of microbial water content
of dust or dirt that fall on the object being sterilized.
These spores come either from the air or from the
As discussed earlier, water has a direct influence on the
activity of persons working near the object. The naked
resistance of microorganisms to dry-heat destruction.
microbial cells of the first group are generally easily
The destruction rate of spores is a function of the
killed by dry-heat sterilization, but the spores found
quantity of water in the cell at the time of heating. This
in soil particles can be difficult to kill. Recent findings
water content is only constant under certain conditions
on the dry-heat destruction characteristics of micro-
and in most conditions, the moisture content of the cell
flora associated with soil particles include the following:
can change so that the secondary variables cause con-
fusion in analysis of results. The water vapor pressure
1. Microorganisms found in soil are widely vari-
in the atmosphere surrounding the cell determines the
able, having different survival characteristics.
movement of water to or from microorganisms on sur-
2. Different organisms found in soil are highly
faces. Research found that when the humidity in air
resistant to dry heat, including Bacillus xero-
passing over spores was increased from 0–0.2, the D
thermodurans, which was found to have a
value also increased by a factor of 100. Spores of inter-
D125
C value of 139 hr.
mediate moisture content with an RH between 0.1 and
3. Some organisms can become more resistant to
0.6 were found to be more resistant to the effects of
heat when encapsulated in particles, such as B.
heat at temperatures between 100
C and 135
C than
subtilis.
spores of either greater or lesser moisture content.[8]
4. It is estimated that 1 spore in 103 and 105 is
extremely resistant to dry heat when found in
Opened and closed systems
soil.
5. The bound water in soil particles may over-
Opened and closed systems were first suggested to indi-
shadow the effect of the humidity level on the
cate the relative control of the heating environment on
destruction rate of the spore.[8]
water loss or gain of spores.
In an open system, the atmosphere surrounding the
The possible variation in numbers and species of
spore determines the spore water content. In this sys-
microorganisms found in soil and the physical con-
tem, cells are heated, and during this heating time,
ditions of their location are unlimited. Spores found in
water may be lost or gained by the cell. The cells can
soil can be located at any point on or within the particle.
gain or lose water either rapidly or slowly, depending
Spores can become completely encapsulated in soil
on the physical system and the nature of the spore.[8]
particles through the various soil wetting and drying
In a closed system, the spore water content is not
cycles, which can increase resistance by a factor of 10.[8]
influenced by the heating environment, but is more a
function of condition inside the enclosure. Closed sys-
tems are different in that water movement and water Destruction of Endotoxins
availability to the cell are restricted. The quantity of
water that is initially present in the enclosure limits Bacterial endotoxins, or pyrogens, are substances that
the quantity of water that is available or that can move will cause a variety of symptoms such as a rise in body
to or from the cell. Initial water content and enclosure temperature when injected in large amounts into
volume of the closed system are two important para- human or animal bodies. In pharmaceutical products
meters. The relative humidity of the atmosphere in and medical devices intended for parenteral injection,
the enclosure at the time of sealing determines the the absence of endotoxins has equal or greater
water concentration in the cell during heating as well importance than sterility. None of the other large-scale
Sterilization: Dry Heat 3517

Sterilization
Statistical–
sterilization methods, such as saturated steam, gamma reductions is between 3 and over 66 hr. At 250
C, six
radiation, and ethylene oxide, are capable of destroy- log10 reductions have been demonstrated. For this rea-
ing endotoxins.[6] son, depyrogenation cycles need to be developed and
All microorganisms seem to be capable of produc- validated empirically. However, the USP recommends
ing pyrogens, but the most potent forms of pyrogens a temperature of at least 250
C.[6]
are associated with gram-negative bacteria. Located
externally to the peptidoglycan cell wall of gram-
negative bacteria is a loosely structured envelope. DISADVANTAGES OF DRY-HEAT
The outer layer of this envelope is made up of lipopo- STERILIZATION
lysaccharides linked to phospholipids and proteins that
act as a permeability barrier that stops the diffusion of Dry heat should be used only for materials that cannot
exoenzymes into the greater environment. It is the lipo- be sterilized by steam either because the moisture
polysaccharide fractions of the cell envelope that have would damage the materials or they would be imper-
been shown to stimulate the pyrogenic response and meable to it. There are many complicating factors
are termed bacterial endotoxins. Purified endotoxin, associated with dry-heat sterilization. The process of
which consists only of lipopolysaccharide, is pyrogenic steam sterilization is accomplished by saturated mois-
in lower doses than naturally occurring endotoxins. ture; in dry-heat sterilization, the moisture can vary
There are three distinct chemical layers of lipopolysac- considerably. Because of the loss of moisture, the death
charide; they include an inner core called lipid A, an rate of spores might change with the continued
intermediate polysaccharide layer, and an outer poly- application of heat.[1]
saccharide side chain. Lipid A is the region that is Although air is the least expensive means of sterili-
responsible for pyrogenicity and consists of a highly zation, it is not the best heat transfer medium. In oven
substituted disaccharide of glucosamine.[6] sterilization, there are several drawbacks. One draw-
The dry-heat destruction of endotoxins is complex back is the lack of uniformity of temperature within
and poorly understood. Most evidence has shown that the oven. Hot air has a tendency to stratify and poorly
destruction follows second-order kinetics with an penetrate masses of cooler material, but moving the air
initial high rate of destruction followed by a slower stream can accelerate heat transfer. Slow heating and
terminal rate.[6] The kinetics of dry-heat destruction cooling down because of low heat transfer rates from
for lipopolysaccharide are expressed in terms of D1 air to product is another problem with ovens that leads
and D2, for the initial and secondary first-order reac- to longer sterilization cycles. Other disadvantages of
tion rates, respectively. Previous research found that dry-heat sterilization can occur because of the high
there was greater reduction in the dry-heat resistance temperatures, which can cause heat damage or even
of lipopolysaccharide in whole cells than that in the charring of materials. Materials can also be damaged
semipurified state. The D1 value for the whole cells due to oxidation because the medium that facilitates
was found to be 1.6, and the D2 value was 12. In the this destructive action also augments its deleterious
semipurified state, D1 was 3.7 and D2 was 29.4. Thus effects.[1]
the rate of the destruction was two times faster when
whole cells were used.[9]
There is basically no endotoxin destruction at tem- CONCLUSIONS
peratures below 80
C, and the D values can be as high
as 20 min for dry-heat temperatures of around 170
C. Although much work has been performed to improve
The second-order models give a better estimate of and develop other methods of sterilization, very little
endotoxin destruction kinetics at temperatures has been accomplished in the field of dry-heat steriliza-
above 250
C than in the 170–250
C range.[6] Generally, tion. The process is time consuming and difficult to con-
the following conditions are required for endotoxin trol because of the temperature stratification and slow
destruction: heating rate.[10] Dry heat is still the agent of choice for
sterilizing items that might not be adequately penetrated
 230
C—60 to 90 min. by steam and that will tolerate high temperatures, such
 250
C—30 to 60 min.[5] as oils, petrolatum, and closed containers.[2]
The development of the infrared radiation tunnel in
The USP and FDA both state that a claim to depy- recent years has opened new possibilities for using dry
rogenation should be supported by evidence that any heat in high-temperature short-time sterilization pro-
endotoxin present on an item prior to treatment cesses.[4] The main advantage of dry-heat sterilization
has been inactivated to no more than 1/1000 of the is its penetrating power. It is not as corrosive as steam
original amount or a three log10 reduction. At 170
C, for metals and sharp instruments, and it does not
the minimum time required to obtain three log10 erode ground glass surfaces, which allows glass to be
 3518 Sterilization: Dry Heat
Sterilization
Statistical–

sterilized at much higher temperatures for shorter 5. Remington, J. Dry heat treatments. In The Science and
periods of time.[1] Pharmacy; Mack Publishing Company: Easton, PA, 1995;
1472–1474.
6. Halls, N. Dry heat sterilization and depyrogenation. In
Achieving Sterility in Medical and Pharmaceutical
REFERENCES Products; Marcel Dekker, Inc.: New York, 1994;
109–120.
1. Joslyn, L. Sterilization by heat. In Disinfection, Steriliza- 7. Bruch, C.W. Levels of sterility: probabilities of survivors vs.
tion, and Preservation, 3rd Ed.; Block, S., Ed.; Lea and biological indicators. Bull. Parenter. Drug Assoc. 1974,
Febiger: Philadelphia, 1983; 3, 27–30, 766–767. 28 (3), 105–113.
2. Validation of Dry Heat Processes Used for Sterilization 8. Block, S.S. Disinfection, Sterilization, and Depyrogenation,
and Depyrogenation, Technical Report No. 3; Parenteral 4th Ed.; Lea and Febiger: Malvern, PA, 1991; 35, 97–101,
Drug Association: Philadelphia, 1981. 514–515.
3. Kupiec, T.; Mathews, P.; Ahmad, R. Dry-heat sterilization 9. Tsuji, K.; Harrison, S. Dry-heat destruction of lipopolysac-
of parenteral oil vehicles. Int. J. Pharm. Compd. 2000, 4 (3), charide: Dry-heat destruction kinetics. Appl. Environ.
223–224. Microbiol. 1978, 36, 710–714.
4. Molin, G.; Molin, O. Dry-heat sterilization of pharmaceuti- 10. Molin, G.; Östlund, K. Dry-heat inactivation of Bacillus
cal glassware using hot air or infra-red radiation. Acta subtilis var. niger spores with special reference to spore
Pharm. Suec. 1976, 13, 476; 481–483. density. Can. J. Microbiol. 1976, 22, 359.
 Sterilization
Statistical–
Sterilization: Ethylene Oxide
Terezinha de Jesus Andreo Pinto
Department of Pharmacy, University of Sao Paulo, Sao Paulo, Brazil

INTRODUCTION sterilant. It is registered as a pesticide by the Environ-

mental Protection Agency (EPA) and is widely used
The biological activity of ethylene oxide (EtO) was as a fumigant in the food industry. It is also used as
initially observed by Cotton and Roark,[1] who a chemical in the manufacture of detergents, plastics,
detected it as an insecticide, in concentrations ranging fibers, film, antifreeze, and other products.[10]
from 3.2 mg L
1 to 32.0 mg L
1. In 1937, Gross and
Dixon[2] requested the patent of EtO as a sterilization
method due to the considerable experimental evidences
MECHANISM OF ETHYLENE OXIDE
it presented.
STERILIZATION
The first EtO commercial application was related to
spices sterilization and/or fumigation.[3–7] Phillips and
The activity of EtO, similar to many other disinfec-
Kaye[8] conducted a series of studies using EtO, focus-
tants, preservative, and sterilizing agents, such as
ing on the sterilization of material with thermal resist-
formaldehyde, beta-propiolactone, methylbromide,
ance problems.
and ethylenimine, depends on an alkylation reaction.[9]
Although its posterior industrial applications included
This reaction occurs with some groups within the
pharmaceutical and cosmetic products, packaging mate-
complex enzymes, proteins, and nucleic acids in the
rials and raw materials of biological origin,[9] EtO is pre-
bacterial cell.[11] These compounds can then no longer
dominantly used in the sterilization process of medical
be effective or necessary to the vital processes of the
devices, such as intravenous sets, cardiopulmonary and
microorganism cell. Furthermore, one would expect
anesthesiology devices. In the hospital environment, the
these effects to vary according to the extensions of
EtO largely contributed as one of the main agents
reactions: static, mutagenic, or toxic.
responsible for the controversial increase in the reutiliza-
The effects of EtO concentration and process tem-
tion of single-use medical devices.
perature were widely studied.[8,12,13] Ernst and Shull[13]
initially verified that the lethality kinetics of Bacillus
ETHYLENE OXIDE CHARACTERISTICS subtilis var. niger spores is of zero order at high levels
of EtO. With the EtO concentration reduction, the
EtO (oxirane, epoxyethane, and dimethylene oxide) is reaction becomes first order. Its sterilization curves
a colorless gas condensing at low temperatures to a at different temperatures show the required EtO con-
mobile liquid. It is miscible in all proportions with centration, and the duplication of the reaction rate of
water, alcohol, ether, and most organic solvents. Its every 10 C.
vapors are flammable and explosive. The physical It has been well established that microorganisms
properties of EtO are summarized in Table 1. that have been equilibrated with atmospheric
EtO is highly reactive; industrially, it is a primary humidity—or even dried to very low humidity, how-
chemical intermediate for a wide variety of compounds. ever, retaining some free water—are killed easier than
The three-membered ring is opened in most of the reac- those desiccated. It is accepted that organic chemical
tions, which give a series of polyethylene glycol deriva- reactions occur through an activated complex forma-
tives of increasing chain length and water solubility. tion; thus, we can infer that water influences this acti-
Ethylene’s strained configuration has been a subject vation. Water must also be present as a reaction
of bonding and molecular structure studies. It is gener- medium or solvent if biological entities are to be
ally non-corrosive to metals and leaves no odor or taste ionized, so that they can enter a transition state with
residues on the materials submitted to its contact. The EtO.[14] Furthermore, no sterilizing activity can be
vapors of EtO (not the liquid form) can be flammable observed when a non-polar solvent such as dioxane
and explosive at concentrations above 3% in air. For and chloroform replaces water.
this reason, it is often combined in sterilant mixtures Water has another function in this sterilization
with an inert gas to eliminate flammability. process—the sterilizing agent concentration on the sur-
EtO is used outside the health care industry as well. face of the microorganism. One can visualize micro-
Actually, only a small fraction of EtO is used as a organisms in an EtO environment in which all the
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-120012010
Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved. 3519
 3520 Sterilization: Ethylene Oxide
Sterilization
Statistical–

Table 1 Typical physical and chemical properties of ethylene oxide

Molecular weight 44.05
Specific gravity (H2O ¼ 1) at 0.8700
68 /39.2 F (20 /4 C)
Specific gravity (air ¼ 1) 1.49
 
Vapor pressure at 68 F (20 C) 22 psia (151.7 kPa abs)
Solubility in water Complete
Percent volatile by volume 100.0
Evaporation rate (butyl acetate ¼ 1) High
Boiling point at 1 atm 50.9 F (10.5 C)
Freezing point at 1 atm
170.7 F (
112.6 C)
Appearance, odor, and state Colorless liquid and gas at normal temperature and pressure.
Non-residual, ether-like odor about 500 ppm. Odor not detectable
until well above the PEL. Shipped and stored under nitrogen
pressure as a liquid

sterilant is in the vapor phase, and in which the reac- sampling period of 15 min. In addition to these regula-
tion rate is dependent on both the number of collisions tory pressures, high levels of concern about workers
of EtO molecules with active sites of the organism, and protection and the potential liability implications
the colliding molecules absorption by the organism, have significantly increased industry’s interest in control
followed by migration of the EtO molecules to the measures for EtO.
active sites. The collision reaction is quite slow; how- The main EtO sources that potentially contribute to
ever, in an atmosphere containing water vapor and workers’ exposure in sterilization plants generally can
EtO, water will equilibrate the organism in the liquid be divided into four major categories.[18]
phase, dissolve the EtO, and concentrate the EtO on
the organism as a solution, thereby increasing the rate  Exposures associated with sterilization equipment
of microorganism killing.[14] operation: sterilizer (door gasket leaks, piping
leaks, leaks from valves and fittings, vent line leaks,
and plume recirculation); vacuum pump (seal leaks,
TECHNOLOGY OVERVIEWS
degassing from sealant discharged to drain); steri-
lant gas storage and delivery system, such as tanks,
Occupational Engineering Considerations
cylinders, or drums (leaks from valves and fittings,
gas line leaks, and releases during change over).
Hazard assessment of EtO has involved considerations
 Exposures associated with sterilized product degas-
about its chemical composition, characteristics, and
sing: unloading, transfer, storage, and spore sample
reactivity. In spite of EtO’s readiness to be trans-
removal.
formed in the body to ethylene glycol and ethylene  Exposures associated with maintenance opera-
chlorohydrene, and its biological half-life of about
tions: routine maintenance and repair, preventive
10 min, it is distributed throughout most body organs.
maintenance.
Studies with cells, animals, and also epidemiological
 Exposures associated with emergency situations:
ones, showed that EtO is able to produce neurophar-
major leaks, major spills, and equipment malfunctions.
macologic, neurotoxic, reproductive, teratogenic, and
mutagenic effects.[15–17]
The Occupational Safety and Health Administra- Engineering solutions must be studied, developed,
tion (OSHA) is responsible for regulating the in-plant and introduced in order to eliminate or, at least, mini-
occupational exposure to EtO in the United States. mize problems in all the steps.[19]
Under the current OSHA standard, accepted by
several other countries, the permissible exposure level Ventilation
(PEL) of EtO is 1 ppm (1.8 mg m
3) per 8 hr (time-
weighted average). However, an action level of General area ventilation systems are designed on the
0.5 ppm is advisable, since it is a guarantee of employ- basis of how much EtO can be degassed into the spe-
ers from low employee exposure levels. cific area mentioned. It is essential to consider the
The OSHA has also established an exclusion limit for total quantity of EtO remaining in the product after
EtO of 5 ppm (9.0 mg m
3) average over a maximum sterilization. This will include packaging EtO in the
Sterilization: Ethylene Oxide 3521

Sterilization
Statistical–
cardboard vapor spaces and the primary packaging, the sterilizer must have double doors and one must
as well as the absorbed/adsorbed EtO in cardboard, provide automatically taking out the pallets from the
the packaging, the product, and the pallets. sterilizer and transporting them into the tunnel.

Vacuum pump modifications Automated systems

Vacuum pump modifications usually involve conver- The most efficient way to protect the worker is keep him
sion of a once-through liquid ring pump to a full seal- away from all operations, like loading, unloading, and
ant recovery pump. This will eliminate any continuous starting procedures during the cycle. So the sterilization
liquid dischargement to the drain. The sealant can process should be automated and computerized.[20]
either be water or oil. The use of water places limita- Besides the worker protection, the computerized system
tions on the maximum vacuum depth, which is reason- allows the cycles ‘‘self-certification,’’ which adds
ably attenuated because of the vapor pressure effects. reproducibility advantages to efficiency of the steriliza-
The use of oil eliminates this maximum vacuum depth tion cycle.
limitation. Fig. 1 (derivada do 6, mas modificada) shows a plant
design that exempts the worker presence, except near
the control panel. The residue monitoring should be
Hot degassing
included in the system. It must embrace the environ-
ment and obviously all processed products and dis-
The major source of worker exposure to EtO in the
charges. Among several options, gas chromatography,
plant is the degassing operation after removal from
especially the one with head space, has been the most
the sterilizer. Different types of products absorb or
adjusted.
entrap EtO at different levels, depending upon the nat-
ure of the product, its specific materials of construc-
tion, how it is packaged, and the specific sterilization Ethylene Oxide Emission Control
cycle to which it was exposed. Unfortunately, not all
the absorbed or entrapped EtO is removed in the steri- During the handling of the ambient controls, the dif-
lizer post-evacuation cycle. Hence, the remaining EtO ferent forms of EtO emission must be controlled, in
will degas after the products are removed from the order to minimize indirect contamination to the popu-
sterilizer. This degassing process typically takes place lation. For example, according to the amendment of
over an extended period of time (days) at typical plant the TA-Luft (German air pollution regulations) in
ambient conditions and usually it is necessary that the 1986, the emission limit of EtO has been fixed to a con-
products be quarantined for this period. centration of 5 mg N
1 m
3 at a mass flow >25 g hr
1.
The use of heat in hot degassing chambers or rooms Mayer et al.[21] described an appropriate plant of com-
to accelerate the EtO degassing has been known for pact design that, using installations for ethylene glycol
a long time. A hot degassing tunnel built directly reactions, ensures these limiting values for the steriliza-
behind the sterilizer has also been used. In this case, tion with EtO.

Pressure gas

Weighing gas
Pressure
Relative humidity
Internal temperature
Chamber temperature

Chamber
Information
input

Gas Unit control

cylinders

Screen
Emission
control
Scale
Fig. 1 Example of a steriliza-
Injection tion plant provided with auto-
Printing
control matic system.
 3522 Sterilization: Ethylene Oxide
Sterilization
Statistical–

There are current technologies that can be used to lower temperature (204–426 C or 400–800 F). The
control EtO emissions to the environment, with specific mixture is then passed through a catalyst bed where
advantages and disadvantages.[22] EtO is heated with oxygen and converted to carbon
dioxide and water.
Even with the advantage of high EtO oxidation
Water scrubbing
efficiency, there are limitations related to potential
decomposition of mixture compounds in toxic by-
This technology involves the absorption of EtO into
products (such as phosgene). In addition, the equip-
water in a packed column. The gas stream discharged
ment cost is relatively high.
from the sterilizer enters the column near the bottom
and flows upward through the packing material. There
are systems in which absorption of gaseous EtO into
the aqueous solution occurs, with its posterior reaction TECHNOLOGY NEW TRENDS
with water to form ethylene glycol, promoting cooling
and recycling of the solution for further EtO removal. Related to explosive decomposition properties of EtO,
The use of an acid reaction (0.1 N is a good concen- pure liquid EtO can inflame in the presence of an
tration, Fig. 2) with water is a good option; anyway, it ignition source.[23,24] The precise threshold limits for
will be necessary to neutralize water, before its discharge liquid decomposition are influenced to some extent
to the environment (usually once or twice a year). by the type of ignition source, as well as by the
geometry of the vessel used. Pure EtO vapor can
H 2 O4 explode by decomposition in the presence of common
C2 H4 O þ H2 O
! HOC2 H4 OH þ DðheatÞ
igniters. Pure EtO vapor at normal storage conditions
is more difficult to ignite than mixtures of EtO and air
or mixtures of hydrocarbons and air. The potential
Thermal incineration
for decomposition can be eliminated by diluting EtO
vapor with a specified proportion of inert gas.
Thermal incineration uses a flame to oxidize EtO to
Though EtO itself may be used to carry out the ster-
carbon dioxide and water at high temperature (typi-
ilization, since the beginning of its use as a sterilant,
cally 760–982 C or 1400–1800 F). A residence time
this has not been done. Instead, EtO sterilant has been
of 0.3 sec–0.5 sec is enough to achieve a high EtO
most used in a mixture with a flame retardant. Over
destruction efficiency.
the 1970s and 1980s, dichlorodifluoromethane, known
Difficulties are related to the high cost of the equip-
in the industry as CFC 12, was the flame retardant of
ment and to limitations in the case of mixtures, when
choice for use with EtO in a sterilant mixture. The
hydrogen chloride (HCl) and hydrogen fluoride (HF)
most commonly used mixture consisted of 27.3 mol%
are involved.
(12 wt%) EtO and 72.7 mol% (88 wt%) CFC 12; this
mixture is commonly referred to 12–88 in the industry.
Catalytic oxidation Also, a 10 : 90 mixture of EtO : carbon dioxide (CO2)
was used, but requiring an increasing of 15 psig in the
In a catalytic oxidation system, the gas stream exhaus- sterilization pressure (this mixture contains only 10%
ted from the sterilization is first diluted with relatively EtO in volume, in contrast to 27% in volume of the
large quantities of air and heated up to a considerably 12 : 88 mixture).[10]

100
90
Reaction time (min)

80
70
60
50
40
30
20
10
0
0.1 1 5 18 28
Acid concentration (N)

Fig. 2 Reaction time to form ethylene glycol considering time.

Sterilization: Ethylene Oxide 3523

Sterilization
Statistical–
Vent to
outside Heated gas
to chamber
Inert gas
Pneumatic valve
Inert gas Manual valve
Pressure regulator valve

ETO Orifice flange

Retention valve
Low ETO Filter
detector

ETO
ETO
(OPTION) Liquid to
gas
Ground Ground vaporizer
cable
Pure Pure cable Temperature
ETO ETO controlled
Drum Drum
#1 #2

Fig. 3 Example of a single installation for a 100% EtO sterilization system.

Due to problems that have arisen with the use of maintained in the 100% EtO sterilizer, so that if any
CFC12 (one of the compounds which causes significant leaks occurred, outside air would be sucked into the
damage to the ozone layer in the upper atmosphere), sterilizer, rather than EtO would leak out, as can occur
worldwide reduction and elimination of CFC12-use with positive sterilizer pressures.
have been started up. This restriction, although related Nowadays, it is known how to use 100% EtO along
to inert dilutor and not to EtO, reached great part of with an inert gas like nitrogen to get safe sterilization,
the users being thus added to the occupational limits even for the most sensitive packaging. Pressures may
requirements. The new technician-scientific concepts be balanced and a modern computer that controls
reached the economic area[25] aggravating extensive 100% EtO sterilizer is a very versatile piece of equip-
fetching of alternatives. ment. Fig. 3 shows schematic installations with an inert
Alternatives that had gained new impulse were the gas hookup to the drum. There are also two devices to
mixtures with carbon dioxide. The 30 : 70 mixture with make sure that the EtO supply to the sterilizer is shut
inflammability features has led to the construction of off when the drum is emptied. One is the weight scale,
new plants, considering the requirements. which automatically signals the computer when a pre-
Another option is a non-flammable EtO—carbon set amount of EtO has been withdrawn and shuts off
dioxide mixture that contains less than 40% of the the drum by a compressed air valve. The another
EtO per unit volume, as does 12–88. Thus, sterilization device is the computer that stops the flow when it
must be carried out either at higher pressures as for senses a change from EtO to the inert gas. Without
longer exposition times. Furthermore, the large differ- these two devices, it is possible that the drum could
ence in the vapor pressures of EtO and carbon dioxide be filled with inert gas and, of course, this would not
causes the mixture to separate upon withdrawal from sterilize the materials.
the storage tank or cylinder, raising the danger of deli- Of course, there are no electrical switches or an
vering a sterilant mixture rich in carbon dioxide, which ignition source in the closed sterilizer–heat exchanger
will not sterilize, or rich in EtO, which is explosive. system. Even the presence of high amounts of moisture
Several companies also re-evaluated their positions both in the product from the preconditioning rooms
about using 100% EtO.[25] In fact, the high costs to and from the humidity injection system would reduce
build an explosion-proof equipment with adequate the likelihood of an explosion by absorbing the EtO
installations would be paid back in savings by switch- into the moisture and by eliminating the cause of
ing to 100% EtO. Also, in the past, the main difference static charges.
between sterilization with 100% EtO and 12/88 was If inert gas lines are hooked into the EtO pipes near
that a deep vacuum was generally used for 100% EtO the EtO source, inert gas can sweep the EtO from the
sterilization, so that all of the air in the package was lines so that they will be either filled with the inert
sucked out, then replaced with a mixture of EtO and gas or be empty. Inverting the filters and flushing the
moisture; this deep intense vacuum required breath- lines with inert gas can keep any EtO from being col-
able packaging material or vents that would allow lected in the filter housing—that also eliminates the
the air inside to escape, so the package would not blow exposure involved in changing filters. Filter changing
up like a balloon and break. A vacuum was generally is probably the major source of exposure for all types
 3524 Sterilization: Ethylene Oxide
Sterilization
Statistical–

of EtO sterilization and this system virtually eliminates contaminants on carriers.[27] Also, a study performed[28]
that problem.[22] to compare sterilization effects between OxyfumeÕ 12
The use of 100% EtO sterilization techniques offers (using EtO and CFC12, 12 : 88) and Oxyfume 2002
low residuals due to the more efficient EtO removal (using EtO and HCFC 22 and 124, 10 : 63 : 27), under
from the package by the deep vacuum. The process different concentrations (450 mg L
1 and 600 mg L
1)
of ‘‘pulsing’’ the vacuum as reported by Christensen[26] and different temperatures (45, 55, and 65 C) revealed
not only greatly lowers residuals which form glycols similar results, even in different concentrations (similar
and chlorohydrins by quickly reducing the amount of D value using Bacillus subtilis, var. niger, ATCC 9372).
EtO in the package, but also allows degassing outside Additional advantages of this new mixture are
the sterilizer to be done within the OSHA limits. In related to the same equipment and installations pre-
fact, one of the latest trends is to do all the degassing viously developed for the CFC mixture (Fig. 5), with
in the sterilizer, so OSHA exposure levels are never only a little increase in pressure or time of exposition,
even approached. This procedure is ideal for 100% during the sterilization cycle. The only concern is that,
EtO systems, especially those having an EtO disposal at present, HCFC is targeted to be phased out by the
unit (Fig. 4). year 2020 in Canada (2 in 9) and 2030 in the United
Recent knowledge about ozone-depleting gases and States (3 in 9).
an international consensus on the need of reducing Different options can require other kinds of equip-
their effects promoted a search for alternative chemi- ment, designed with the goal of protecting the workers,
cals to several different CFC applications. From these, environmental preservation and safety of products.
one of the most interesting are the hydrochlorofluoro-
carbons (HCFCs) which, besides being similar to CFC
in inhibition of explosiveness and inflammability of Validation and Routine Control of
EtO, can also be used as transitional compounds while Ethylene Oxide Sterilization
more environmentally suitable compounds are not
available. An EtO–HCFC (10 : 90) mixture was com- Even though the definition of sterility is an absolute
pared to 100% ethylene sterilizing gas to determine condition, the effectiveness of the sterilization process
its relative ability to kill seven different bacteria. can be determined by measuring the reduction of
Results demonstrated that the EtO–HCFC mixture microbial population. Such measurements reveal the
was equivalent to the 100% EtO sterilant to kill kinetics of microbial inactivation, and it is from the
vegetative organisms, as well as spore suspensions in exponential nature of inactivation that the concept of
the absence of serum and salt, and better with these sterility assurance level (SAL) is derived. This value

1. Pre vacuum - Pre heat

2. Humidification - 1,2,n...
3. Gas exposure
4. Final vacuum - Fresh air or nitrogen purge
Pressure 5. Cycle complete automatic aeration begin

Post vacuum
Initial vacuum
Atmosphere Gas addition Exhaust Air or nitrogen wash

Exposition

Additional air wash

must be used

1 2 n...

Humidification &
conditioning

1 2 3 4 5

Fig. 4 A typical sterilization cycle observed with single-use 100% EtO cartridges.
Sterilization: Ethylene Oxide 3525

Sterilization
Statistical–
Load Humidification & Exhaust
sterilizer conditioning Post vacuum
Close Gas Air or nitrogen wash
Exposition
sterilizer addition 2nd vacuum
Sterilizer door
2nd air wash
pressure

Initial Additional
vacuum air wash
may be used
2

Atmospheric
pressure
3 4 3 4

1
Vacuum

1. Presterilization conditioning
2. Sterilization
3. Evacuation
4. Air or nitrogen wash

Time line

Fig. 5 Typical sterilization cycle with CFC–EtO and HCFC–EtO mixtures.

is expressed as a negative power to the base 10. SALs The adoption of the validation standard document
of both 10
3 and 10
6 have been adopted worldwide. reflects the medical industry ongoing commitment to
However, there is a tendency to adopt a single safety, efficacy, and global harmonization of manufac-
standard, 10
6.[29] turing standards. Its purpose is to provide confidence
The validation of a sterilization process is a docu- in all markets that the sterilization process is properly
mented procedure, which obtains records and inter- designed, tested, and monitored.[29]
prets the data required, to show that a process will According to the standard, the sterilization process
consistently comply with predetermined specification. using EtO shall include preconditioning and/or
Besides respecting the SAL, the biocompatibility and conditioning, sterilization cycle, and aeration. Precon-
functional properties of a product have to be respected. ditioning and/or conditioning treatments must be per-
The most important current standard is ANSI/ formed under controlled conditions for a defined
AAMI/ISSO 11135—1994 (revision of ANSI/AAMI period of time to achieve specified temperature and
ST27—1988). Medical devices validation and routine relative humidity within the load. The sterilization
control of EtO sterilization were approved by the cycle includes: air removal, conditioning (if used), ster-
Association for the Advancement of Medical Instru- ilant injection, maintenance of specified conditions for
mentation (AAMI), the American National Standards the exposure time, sterilant removal, flushing (if used),
Institute, Inc., (ANSI) and the International Organiza- and air admission to atmospheric pressure. Aeration
tion for Standardization (ISO). This standard consid- must be performed with product retained under speci-
ers the following steps for validation: commissioning fied conditions, in the sterilizer and/or in a separate
and performance qualification, which includes physical chamber or room. One of the most important differ-
and microbiological qualification. Medical devices to be ences is that ISO 11135 requires additional temper-
sterilized shall be manufactured under conditions that ature and humidity monitoring of the load, both
ensure that their bioburden is consistently low. Another during validation and routine.[32]
important point is about the AAMI specification that According to the ISO standard, the temperature
the biological indicator evaluator resistometer (BIER) and humidity of the load must be measured during
is the appropriate vessel for evaluating resistivity to the performance qualification. These measurements
sterilization of biological indicators.[30,31] should be used to establish range specifications for
 3526 Sterilization: Ethylene Oxide
Sterilization
Statistical–

the temperature and humidity of the product at the end the lethality shall be determined by the construction
of both the preconditioning and conditioning steps, of a survivor curve using direct enumeration of a sur-
before adding the sterilant into the chamber. Only in vivor. At least five points employing graded exposure
an Annex of ISO standard (B), included as a guidance, times to EtO, with all other process parameters
at the end of preconditioning, the measured tempera- remaining constant, except time, shall be included on
ture and humidity ranges within the sterilization load the survivor curve. The second possibility is the
should not exceed 5 C and 15% humidity. This fraction-negative method, in which indicators for
annex also contains the information that a recorded EtO sterilization are also exposed to EtO in graded
temperature range within an empty chamber during exposure times with all process parameters remaining
sterilant exposure of less than or equal to 3 C of constant, except time. After exposure, the indicators
the required set point should be obtained. for EtO sterilization shall be tested by direct immersion
Throughout the exposure time, the sterilization load in an appropriate culture medium. Both methods per-
should attain the minimum specified temperature and mit calculation of D value (Fig. 6).[35] The method C,
the temperature range across the product load should or half-cycle method, can be used only for conven-
be less than or equal to 10 C at any given time during tional product release. It involves the determination
sterilant exposure. The actual temperature should be of the minimum time of exposure to EtO at which
determined during physical performance qualification. there are no survivors, with all other process para-
Although temperature measurements have long been meters,[21] except time remaining constant. Two further
required, direct humidity measurement of the load experiments should be performed to confirm the mini-
has not. Manufacturers have used vapor pressure cal- mum time. Both should show no growth from biologi-
culations to ensure that when steam is added to the cal indicators. The specified exposure time should at
chamber after initial evacuation, the resulting increase least double this minimum time.
in pressure indicates attainment of a level of humidity Considering the parametric process, it requires more
appropriate to support microbial inactivation. attention in comparison with conventional release. In
By interrupting the sterilization cycle just before the this case, a secondary measure of chamber temperature
introduction of EtO gas, validation runs can be per- is required, and the chamber humidity and gas concen-
formed using currently available chamber humidity tration must be directly monitored, what constitutes an
sensors, even though, care maintenance and calibration important challenge, mainly related to safety in 100%
of these sensors are very important, and a real chal- EtO cycles (Norma ISO). An inherent characteristic
lenge is to improve their state-of-art. of the sterilization process that reduces the interest in
A double check for EtO concentration is required obtaining conditions that allow the parametric liber-
during sterilization (weight of sterilant, volume on ation is related to the attendance to specified limits
direct analyses, besides the pressure rise). During aer- of EtO, ethylene glycol and ethylene chlorohydrene
ation, the temperature of the load must be monitored previously to the product release, what demands
in reference to a specified range established. The micro- additional time.[36]
biological performance qualification should demon-
strate the adequacy of the process to the sterilization
of the product by the inactivation of indicators for
9
EtO sterilization. The bioburden of the product has to
be established, and indicators for EtO sterilization— 8
Log10 microbial population

spores of B. subtilis var. niger, which are being 7

considered for another classification as B. atrophaeus
on the basis of high DNA–DNA reassociation values 6

and confirmatory automated RiboPrint analysis[33] 5

must comply with EN 866-2. These indicators shall
4 Plastic
be placed at representative positions throughout the
sterilization load under the cycle conditions selected 3
Aluminium
to deliver less lethality. The indicator should repre- 2
Paper
sent, considering the holder where the spores are
1
inoculated, the load configuration and the package
0 5 10 15 20 25
that protects it in the worst sterilizer conditions. This
Time (min)
is added to the worst conditions in the configuration
of the load.[10,34] Fig. 6 Microbial death rate (D determination using direct
According to the standard, the microbiological enumeration of survivors) for B. Subtilis var. niger on paper
qualification can be performed by the lethality determi- (D ¼ 4310 min), aluminum (D ¼ 6143 min), and plastic
nation of the cycle by three methods. In one of them, (D ¼ 12,300 min) carrier.
Sterilization: Ethylene Oxide 3527

Sterilization
Statistical–
Another possibility, proposed by Rodriguez et al.,[37] 6. Smith, H.W. Treated spices reduce spoilage. Food Ind.
considers the concept of accumulated lethality (Fo), 1940, 12, 50–72.
7. Yesair, J.; Williams, O.B. Spice contamination and its
used for thermal sterilization, but applied to optimiza- control. Food Res. 1942, 7, 118–126.
tion of EtO sterilization technology. A mathematical 8. Phillips, C.R.; Kaye, S. The sterilization action of gaseous
model of the inactivation of biological indicators ethylene oxide. I. Review. Am. J. Hyg. 1949, 50, 270–279.
spores by EtO was developed, along with two formu- 9. Alguire, D.E.; Yeung, A.C. Making cosmetics microbiologi-
las: a ‘‘response’’ equation for calculating the number cally safe. Cosmet. Toiletries 1979, 94, 77–80.
10. Pinto, T.J.A. Aspectos Fundamentais na Validação do
of survivors of a sterilization cycle, and a formula Monitor Biológico Para a Esterilização por Óxido de
for determining the accumulated lethality of exposure Etileno. Tese de Doutorado. Faculdade de Cie ^ncias
to EtO. Experiments verified that the equations are Farmace ^uticas—USP, São Paulo, 1991; 203 pp.
11. Lencioni, E.; Panzarasa, L. La sterilizzazione con ossido di
applicable to processes with relative humidity values etilene. Boll. Chim. Farm. 1977, 116, 378–392.
between 15% and 90%, enabling users to compare the 12. Ernst, R.R.; Doyle, J.E. Sterilization with gaseous ethylene
lethality of dissimilar EtO cycles. oxide: a review of chemical and physical factors. Biotech-
On completion of the validation programme, the nol. Bioeng. 1968, 10, 1–31.
test results should be compiled into a test report, and 13. Ernst, R.R.; Shull, J.J. Ethylene oxide gaseous sterilization.
I. Concentration and temperature effects. Appl. Microbiol.
characterize the certification of validation. 1962, 10, 337–341.
14. Gunther, D.A. The chemistry and biology of eto steriliza-
tion. MD & DI 1980, 6, 31–35.
15. Golberg, L. Hazard Assessment of Ethylene Oxide; CRC
Press: Boca Raton, FL, 1986.
CONCLUSIONS 16. Turchi, G.; Bonatti, S.; Citti, L.; Gervasi, P.G.; Abbondan-
dolo, A. Alkylating properties and genetic activity of 4-
vinylcyclo-hexene metabolites and structurally related
Despite the serious criticisms against EtO related to epoxides. Mutat. Res. 1981, 83, 419
toxicity and environmental aspects, sterilization using 17. Vogel, E.; Natarajan, A.T. The relation between reaction
this gas, 100% or in various compositions, is one of kinetics and mutagenic action of monofunctional alkylating
the most widely used processes. Compared to alterna- agents in higher eucaryotic systems. II. Total and partial
sex-chromosome loss in drosophila. Mutat. Res. 1979,
tive processes, EtO still remains a suitable choice than 62, 101
irradiation, because it promotes molecular alterations 18. Desai, P.R.; Buonicore, A.J. Engineering controls to mini-
in different polymeric compounds, and it also causes mize worker exposure to ethylene oxide at sterilization
facilities. Plant/Oper. Prog. 1990, 9 (2), 103–107.
long-term problems with the ramrods of 60Co, when
19. Gunther, D.A. Evolution of equipment, techniques for
its active life expires. Besides that, the irradiation with industrial EtO sterilization. Am. Perfume Cosmet. 1970,
difficulties persists with accelerated electrons, in spite 85, 35–38.
of the obtained improvements. 20. Lencioni, E.; Franchi, G.; Panzarasa, L. L’Autocertifica-
Furthermore, the EtO process presents wide effec- zione nei processi di degermazione con EtO. Pharm. Acta
Helf. 1987, 62 (10–11), 306–312.
tiveness and possibility of validation in industrial 21. Mayer, V.F.; Agostini, G.; Schaber, K.; Koch, A. Abschein-
sterilizators, when compared to the plasma sterilization lung von ethylenoxide bei sterilisationsprozessen in der
process. It also shows advantages in comparison to pharma industrie. Pharm. Ind. 1989, 51 (3), 294–298.
other sterilant gases (formaldehyde and hydrogen per- 22. Buonicore, A.J.; Desai, P.R. EtO emission control alter-
natives for sterilization facilities. In A Safe Method in
oxide) related to permeability, diffusion, volatilization, Safe Hands—a Practical Approach to Harmonization;
polimerization, and compatibility. EUCOMED Conference on EtO Sterilization, Paris, 1989.
So, since there is not a perfect sterilant agent, we 23. Britton, L.G. Thermal stability and deflagration of ethylene
can consider the EtO process as ideal, although it oxide. Plant/Oper. Prog. 1990, 9 (2), 75–86.
24. June, R.K.; Dye, R.F. Explosive decomposition of ethylene
demands a certain amount of knowledge for its safe oxide. Plant/Oper. Prog. 1990, 9 (2), 64–67.
and effective use. 25. Reichert, M. Low temperature sterilization alternatives—
what are the real costs? Surg. Serv. Manag. 1995, 1 (2),
38–43.
26. Christensen, D.E. Changing EtO sterilizer cycles to reduce
REFERENCES ethylene oxide exposure levels. Med. Dev. Diagn. Ind.
1984, 6, 27–35.
1. Cotton, R.T.; Roark, R.C. Ethylene oxide as a fumigant. 27. Alfa, M.J.; Degagne, P.; Olson, N. Bacterial killing ability
Ind. Eng. Chem. Wash. 1928, 20, 805–809. of 10% EtO plus 90% hydrochlorofluorocarbon sterilizing
gas. Infect. Control Hosp. Epidemiol. 1997, 18 (9),
2. Gross, P.M.; Dixon, L.F. Method of Sterilizing. U.S. Pat. 641–645.
2.075.845, 1937, Apud: Ref. 8.
28. Oliveira, D.C. Esteriliza ção Por Óxido de Etileno: Estudos
3. Hall, I.A. Sterilized spices: new factor in food quality De Efetividade Esterilizante de Misturas Não Explosivas e
control. Food Ind. 1938, 10, 424–425464–467. Compatı́veis com a Camada de Ozo^nio. Faculdade de
4. James, L.H. Reducing the microbial content of spices. Cie^ncias Farmace^uticas—USP, 2000; 159 pp.
Food. Ind. 1938, 10, 428–429. 29. Morissey, R.F.; Bruch, C.W.; Sharbaugh, R.J.; Favero,
5. Jensen, L.B.; Wood, I.H.; Jensen, C.E. Swelling in canned M.S.; Jarvis, W.R.; Masefield, J. Sterility and safety assur-
chopped hams. Ind. Eng. Chem. 1934, 26, 1118–1120. ance of medical devices. MD & DI 1992, 14 (4), 78–81.
 3528 Sterilization: Ethylene Oxide
Sterilization
Statistical–

30. Manning, C.R. Validation EtO packaging/sterilizer config- 35. Paulson, D.S. Calculating D-values for steam sterilization
urations. MD & DI 1990, 1 (3), 52–58. processes. MD & DI 1995, 17 (5), 198–204.
31. AAMI STANDARD. Biological Indicators for EtO Steriliza- 36. Seille, J.M.; Delattre, L.; Meurice, L.; Jaminet, F. Étude de
tion Processes in Health Care Facilities. Estados Unidos, 1986. l’effet d’une sterilisation a l’oxide d’ethylene sur les teneurs
32. Booth, A.F. ISO 1135: new standard presents new chal- residuelles en chlorhydrine du glycol et en ethyleneglycol
lenges. MD & DI 1994, 16 (2), 64–67. dans des articles medico-chirurgicaux a base de PVC, prea-
33. Fritze, D.; Pukall, R. Reclassification of bioindicator strains lablement irradies au cobalt 60. J. Pharm. Belg. 1985, 40 (4),
Bacillus subtilis DSM 675 and Bacillus subtilis DSM 2277 213–221.
as Bacillus atrophaeus. Int. J. Syst. Evol. Microbiol. 2001, 37. Rodriguez, A.C.; Young, B.; Caulk, K.; Zelewski, J.; Kwas-
51, 35–37. nica, S.; Aguirre, S. Calculating accumulated lethality and
34. Pinto, T.J.A.; Saito, T.; Iossif, M. Ethylene oxide steriliza- survivorship in EtO sterilization proceses—the availability
tion: III—influence of carrier nature in a biological monitor of two new equations makes it possible to compare the
performance. PDA J. Pharm. Sci. Technol. 1994, 48 (3), effects of dissimilar process cycles. MD & DI 2001, 5,
155–158. 100–107.
 Sterilization
Statistical–
Sterilization: Moist Heat
Dario Pistolesi
Fedegari Autoclavi SpA, Albuzzano, Italy

INTRODUCTION but they are still being performed. So-called parametric

release (i.e., release based on the evaluation of tightly
To sterilize something means to render it ‘‘aseptic’’ controlled physical parameters of each process, with-
(from the Greek word sepsis, putrefaction, preceded out performing sterility tests) is in fact theoretically
by a privative a). In other words, it means to inactivate accepted by many pharmacopoeias, but is (as stated,
the micro-organisms that may produce this putrefactive for example, in the European Pharmacopeia) ‘‘ . . .
action (in the broadest sense of the term). subject to the approval of the competent authorities
Moist-heat sterilization is achieved when water . . . ,’’ which are generally highly reluctant to grant
vapor (or, more generically, moist heat, i.e., a suitable such approval.
combination of temperature and humidity) at a definite All pharmacopeias consider moist-heat sterilization
temperature is introduced or generated (even indirectly) as the method of choice, i.e., the method to be pre-
at the level of the micro-organisms to be inactivated ferred, unless, of course, the product to be sterilized
and is maintained in such conditions for a definite time. is incompatible with the characteristics of steam. The
As explained in detail hereafter, moist-heat sterilization reason for this preference is the fact that moist-heat
proceeds as an inverse logarithmic progression. There- sterilization provides the best combination of flexibility,
fore, only a treatment of infinite duration provides reliability, and low equipment and operating costs.
absolute certainty that all micro-organisms have been
inactivated.
In pharmaceutical technology, to define an item
OVERVIEW
as sterile, one must be able to demonstrate that on a
statistical basis related to the processing conditions,
Moist-heat sterilization is achieved when a suitable
no more than 1 in 106 units subjected to sterilization
combination of temperature and humidity can be
‘‘may be’’ non-sterile. Therefore, the SAL (Sterility
introduced (or indirectly generated) at the level of the
Assurance Level) of the product must be greater than
micro-organisms to be inactivated. The classic way to
(or equal to) 106. The obvious consequence of this
achieve this is by means of pressurized saturated steam
situation is that although the word sterile expresses
at the temperature of 121
C (250
F). However, other
an absolute concept, the word sterilized, understood
sterilizing media (e.g., superheated water or a steam–
as the result of an adequate sterilization process, has
air mixture) are also frequently used to obviate certain
a probabilistic meaning.
problems that pure steam may pose. Sometimes the
Current pharmacopeias, standards, and guidelines
load is rotated inside the chamber of the sterilizer to
related to sterilization generally use the following type
achieve particular results.
of wording:

. . . Sterilization is a special process because its efficacy

cannot be verified by simple inspection and testing on MOIST-HEAT STERILIZATION KINETICS[3]
the final product . . . . For this reason, sterilization pro-
cesses have to be validated before use, the performance Consider a system contaminated by a single microbio-
of the process monitored routinely and equipment logical species (i.e., an ampoule containing an aqueous
regularly maintained . . . . suspension of a given micro-organism) immersed in
pressurized saturated steam at constant temperature.
Accordingly, installation qualification and oper- One can demonstrate experimentally that the reaction
ational qualification of the sterilizer and validation of of thermal inactivation of the micro-organism develops
the processes, combined with continuous monitoring as a first-order chemical reaction (i.e., as a chemical
of each individual routine process, are now considered decomposition reaction) in which the reaction rate is
fundamental.[1] Therefore, sterility tests (i.e., micro- proportional at all times only to the amount of product
biological tests performed on the final product) have still to be degraded. The proportionality coefficient is
lost much of the significance they had in the past,[2] typical of the species and conditions of the given
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-100000438
Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved. 3529
 3530 Sterilization: Moist Heat
Sterilization
Statistical–

micro-organism. All this seems obvious for dry-heat unit of 100 micro-organisms or 102. If D121 ¼ 1, after
sterilization, but not for steam sterilization, in which 1 min at 121
C, the population will be reduced to
the water vapor molecules would appear to take part 101 ¼ 10. After another minute, only 100 ¼ 1 micro-
in the reaction. Actually, this bimolecular reaction is organism will still be surviving. After another minute,
a first-order reaction, because an excess of steam is the surviving population would be 101 ¼ 1/10
always present and its concentration can be considered micro-organism per unit. In biological terms, such a
constant. contamination is obviously meaningless; statistically,
The most widely used mathematical equation of the it means that there is a probability that 1/10 of the units
above is: of the sterilized batch are still contaminated. Clearly,
after another 5 min of sterilization, this probability will
N ¼ N0 10t=D ð1Þ be reduced to 1/106 or 106. In other words, the SAL
(introduced earlier) is 106.
where N0 ¼ initial number of micro-organisms; A more reassuring SAL, for example 109, is very
t ¼ elapsed time (or sterilization time); N ¼ number often sought. It is sufficient to extend the sterilization
of surviving micro-organisms after exposure time t; for just 3 additional min. The problem, therefore, is
and D ¼ ‘‘decimal decay time,’’ defined as the time evidently not cost-related; rather, it is simply linked
interval required, at a specified and constant tempera- to the risk of subjecting the treated material to thermal
ture, to reduce the microbial population to 1/10 of its degradation.
original quantity. All the above considerations have been made under
At 121
C, the D-values generally oscillate between the assumption that the temperature is kept constant
0.2 and 1.5 min for the various microbial species that during the sterilization period. Obviously, the D-value
can be encountered in pharmaceutical activity. Eq. (1) changes when the temperature changes. When the
allows for two important conclusions: D-values obtained experimentally for a given microbial
species are plotted on a semilogarithmic chart as a
1. the time required to reduce the micro-organism function of the temperature T, a path such as the one
population to any preset value is a function of shown in Fig. 2 is obtained.
the initial concentration, and Clearly, if D is 1.0 at 121
C, it is 0.1 at 131
C and
2. the effect of sterilization in the same conditions 10 at 111
C. In other words, the value of D decreases
(T and t) will be very different according to the or increases by a factor of 10 when the temperature
D-value of the contaminating micro-organism. increases or decreases by 10
C. The algorithm z is
defined as temperature coefficient of moist-heat sterili-
Fig. 1 shows that the same reduction ratio is zation, i.e., the number of degrees of sterilization tem-
achieved for different microbial species (at the same perature that causes a 10-fold variation of D or of the
constant temperature) with an exposure time that is sterilization rate. Depending on the micro-organism
proportional to the D-value of each species.
Consider a batch of units (e.g., a batch of filled
ampoules) with a constant initial population for each 102

102
101
101
Number of m.o. per unit

100
D value

D=2
10−1

10−2 D121 = 1 min

D=1 100
10−3 16
10−4
D = 0.5
10−5 z =10ºC
10−1
10−6 101 106 111 116 121 126 131
0 1 2 3 4 5 6 7 8
Sterilization time (minutes) Sterilization temperature (ºC)

Fig. 1 Death rate curves illustrating decimal reduction Fig. 2 Logarithm of D decreases linearly as temperature
concept. (From Ref.[3].) increases. (From Ref.[3].)
Sterilization: Moist Heat 3531

Sterilization
Statistical–
being considered, the z-value varies between 5 and 15 mixture of steam and air as sterilizing media. These
for the 100–130
C sterilization range. The z-value is processes allow the control of the pressure of the ster-
frequently assumed to be equal to 10 in the absence ilizing medium independently of its temperature (which
of precise experimental data.[1] is impossible to accomplish with pure saturated steam).
It is evident that small temperature variations cause As explained below, these processes are used to treat
dramatic variations in the rate of the sterilization solutions in containers that cannot tolerate the internal
reaction. It is easy to calculate that a variation by only overpressure that is generated inside when sterilized
1
C in the vicinity of 121
C causes a variation with process 1.
of approximately 24% in the value of D, i.e., of the
sterilization rate.
PRESSURIZED SATURATED STEAM METHOD
F0 or Equivalent Time
This is certainly the most widely used and most versatile
moist-heat sterilization method. Accordingly, it is widely
To compare the lethal effect of a sterilization per-
used not only for sterilization of pharmaceutical pro-
formed for any given time tx at any temperature Tx
ducts but also for laboratory and hospital sterilization
(which may vary over the time tx), it is very useful to
and for the treatment of medical devices. Nonetheless,
be able to express this lethal effect by relating it by
it has significant limitations, especially in pharmaceutical
calculation to a given reference temperature. When this
use, which are described later. The sterilizing medium is
reference temperature is 121
C (or, more exactly,
obviously pure pressurized saturated steam. The word
121.11
C, which correspond to 250
F) and z is
saturated means that the steam is in thermodynamic
assumed to be 10 (or 18 if the temperature is expressed
equilibrium with its liquid form (water) at the tempera-
in
F), the resulting algorithm is known as F0 and is
ture being considered.
expressed by:
Typical operating conditions are 121
C (i.e., 250
F)
X for 15 min (or even less); this temperature is matched
10ð Þ
T  121
F0 ¼ Dt z ð2Þ by a saturated steam pressure of 2.05 abs bar (i.e.,
205 kPa). However, higher or lower temperatures (and,
where Dt ¼ time interval between two successive tem- therefore, pressures) are often used, with obvious
perature measurements; T ¼ actual sterilization tem- appropriate adjustments of the holding time.
perature in
C at the time t; and z ¼ temperature The term dry saturated steam is sometimes used. It
coefficient, assumed to be equal to 10. should be made clear that dryness is a theoretical con-
F0 is known as equivalent time because its dimen- dition of steam and that in practice, moist saturated
sion is actually a time expressed in minutes. Clearly, steam is used. This also provides assurance that the
when the values in Eq. (2) are, for example, Dt ¼ 15 min steam really is saturated and is not superheated. The
min and T ¼ 131
C, F0 is 150 min, i.e., the lethal effect use of superheated steam may in fact cause problems
is 10 times higher than that of a sterilization lasting in process management.
15 min at 121
C. If instead Dt ¼ 15 min but T ¼ However, the steam must entrain the smallest pos-
111
C, F0 is equal to 1.5 min, i.e., the lethal effect is 10 sible amount of condensate. The term steam dryness
times smaller than that of a 15 min sterilization at fraction defines the amount of condensate entrained
121
C. An F0 of 12 (delivered to the coldest point of by the moist steam. A dryness fraction of 0.95 means
the load) is generally considered sufficient for adequate that 100 g of moist steam consist of 95 g of dry satu-
sterilization in the pharmaceutical field.[4] rated steam plus 5 g of condensate, which is (or should
be) at the same temperature the steam. A dryness frac-
tion of 0.95 is considered the lower limit of adequacy
MOIST-HEAT STERILIZATION PROCESSES for moist-heat sterilization.
The reliability of sterilization performed by means
Current pharmaceutical production practice uses sub- of saturated steam is based on three essential charac-
stantially three moist-heat sterilization processes: 1) teristics of this medium:
pressurized saturated steam; 2) superheated water;
and 3) steam–air mixture. Process 1 is the traditional 1. When steam condenses, it releases heat at a
multipurpose process, which obviously uses pure constant temperature and in very large amounts;
pressurized saturated steam as sterilizing medium. Pro- 1 kg of steam condensing at 121
C (transforming
cesses 2 and 3 are so-called counterpressure processes; into water at 121
C) releases as much as 2200 kJ
they were introduced in pharmaceutical production (or 525 kcal).
practice approximately 20 years ago and, respectively, 2. The temperatures and pressures of saturated
use a spray superheated water and a homogeneous steam have a two-way correlation. Once the
 3532 Sterilization: Moist Heat
Sterilization
Statistical–

steam temperature is determined, so is its pres- can be intolerable for many types of container.
sure, and vice versa. Saturated steam at 121
C In such cases, it is necessary or convenient to
inevitably has a pressure of 2.05 abs bar; use counterpressure autoclaves (as described
saturated steam at 3.04 abs bar inevitably has later).
a temperature of 134
C. This entails two very
interesting practical possibilities: a pure satu-
rated steam autoclave can be equally tempera- SATURATED STEAM AUTOCLAVES
ture- or pressure-controlled, and regardless of
the parameter used for control, the second Construction
parameter can easily be used to cross-monitor
the first. All sterilizers intended for pharmaceutical use are
3. One gram-molecular weight of water (18 g, i.e., currently made of class AISI 316 stainless steel, includ-
18 ml in the liquid state) as steam at 121
C ing the valves and piping. Other materials may be
and 2.05 abs bar occupies a volume of approxi- acceptable only for service components arranged
mately 15 L. This means that when steam con- downstream of the sterilizer (e.g., the vacuum pump
denses at 121
C, it shrinks in volume by almost or the condensate trap). The service elements arranged
1000 times. Consequently, additional steam spon- upstream of the sterilizer (e.g., heat exchangers or
taneously reaches the material to be sterilized. water pumps) also must be made of stainless steel.
The condensate that forms can be easily removed Silicone rubber or PTFE and its derivatives are gen-
from the autoclave chamber by means of a con- erally used for gaskets (for doors, valves, etc). The
densate trap or by continuous bleeding. chamber of these autoclaves is horizontal, with a
rectangular or (rarely) cylindrical cross-section. The
Apart from these three favorable characteristics, dimensions of these chambers can vary considerably,
other phenomena linked to the use of pure saturated from approximately 100 L to 10 m3 or more.
steam must be considered:

1. To perform its microbiological inactivation Doors

action, the steam must come into contact with
the micro-organisms. This can occur directly Doors are generally rectangular, even though the cham-
or indirectly. It occurs directly when the steam ber is cylindrical. In this case, the doors are inscribed in
makes contact, for example, with a surgical the circumference. There may be one or two doors: two
instrument located in the autoclave chamber. doors are always used when the autoclave leads into
It occurs indirectly when the steam is generated, a sterile room. Two-door autoclaves are often used
for example, inside a sealed ampoule that con- when this requirement does not apply, but the need is
tains an aqueous solution by heat exchange with nonetheless felt to separate the loading area (where
the steam in the chamber. However, it is evident non-sterile products are placed) from the unloading
that it is impossible to steam-sterilize the inside area (where only already-sterilized products can be
of a closed empty ampoule or its contents if placed). The doors may be of various kinds.
they are, for example, an anhydrous oil-based
solution. Side-hinged, manually operated doors
2. The air initially present in the chamber and the retained by radial locking bars
incondensables (generally CO2) possibly entrained
by the steam have molecular weights, and there- At present, these are the most widely used doors. The
fore densities, which are 1.5–2.0 times higher than rim gasket is solid and fixed. The radial bars are
those of steam. Therefore, at the beginning of moved, during closure/opening, by a central hand-
the process, the air must be removed from the wheel that is operated manually. This locking system
chamber, and the steam must not contain incon- requires lubrication, and this can entail microbiological
densables. Otherwise, they tend to stratify in the problems, especially if the door opens onto a sterile
lower portions of the chamber, producing unac- room (the closure method that uses perimetric eye-bolts
ceptable temperature gradients. is now obsolete).
3. When closed non-deformable containers with
aqueous solutions are sterilized, the pressure Vertically or horizontally sliding,
inside can reach values far higher than the cham- automatically operated doors
ber pressure. The reasons for this are explained
in detail later but, in any case, the internal These doors have no lubrication problems, but in the
overpressure can reach or exceed 1.4 bar and horizontally sliding version (which is common for
Sterilization: Moist Heat 3533

Sterilization
Statistical–
industrial-size autoclaves), they have the drawback jacket, circulates inside it, flows out of it, and then
that they occupy a considerable amount of floor space. enters the chamber. The reduction of condensate
The gasket is generally located on the chamber rabbet entrained by the steam can be achieved only with this
and is compressed toward the door by two methods: 1) single-feed approach. 2) Separate-feed in which, of
the gasket is hollow and inflatable (by means of com- course, two controls are provided. The jacket may be
pressed air or steam); and 2) the gasket has a particular fed with plant steam, but the chamber is certainly
dovetail cross-section and is contained in a slot that supplied with ultraclean steam. At present, this is the
also has a dovetail cross-section. An adequate pressure solution generally used for modern autoclaves because
of compressed air in the rear part of the slot is suf- it ensures that no microbiological or particulate con-
ficient to activate the seal on the door, and the release tamination can reach the chamber from the jacket,
of the pressure (without requiring vacuum) is sufficient which is a closed and convoluted space that is practi-
to activate the retraction of the gasket (Fig. 3). cally impossible to clean and inspect accurately.

Diagonally moving doors

Process
These doors combine the positive features of the two
Initial air removal from the chamber
precedidng types. During opening (and, in reverse, dur-
ing closure), the door is automatically moved slightly
The basic reason for air removal has been addressed
upward and laterally, enough to disengage it from
previously. Moreover, loads are often made of porous
the mechanical systems that retain its four sides. Then
or packaged materials, which require reliable and rapid
the operator opens the door manually by turning it
removal of air from the inside of the loads as well.
about its side hinges. The gasket is generally identical
Today, the so-called gravity removal method is used
to the second one described previously.
only for special tasks. In this method, the steam enters
the top part of the chamber and is distributed by a
Jacket suitable sparger (theoretically on a uniform front). The
air escapes through a large valve in the lower part of
Saturated steam autoclaves are generally provided with the chamber by way of two actions: gravity and dis-
a jacket, that is constituted by a second wall that more placement by steam. This method is rather slow and
or less fully encloses the inner chamber and thus forms unsuitable for porous and other loads that may trap
a secondary space around it. For the sake of brevity, the air inside recessed cavities.
various jacket construction methods are not presented A modern autoclave is generally equipped with a
here. The purpose of a jacket is summarized as follows: water-ring pump that can produce a vacuum of approxi-
1) to preheat the autoclave initially and keep it warm mately 70 residual mbar in the chamber. Accordingly,
during loading/unloading; 2) to preheat the load dur- almost 7% of the air is not removed. The following two
ing the initial air removal phase; 3) to contribute to methods are essentially used for completing air removal.
the drying of the load in the final vacuum phase; and
4) to reduce any condensate entrained by the steam. Pulsed Vacuum. Once the maximum initial vacuum
Steam can be fed to the jacket-chamber assembly has been reached, the pump is stopped and steam
by: 1) Single feed in which the steam first enters the enters the chamber until an approximate atmospheric
pressure (or a higher pressure) is reached; then vacuum
is produced again. Three vacuum/pressure pulses are
generally sufficient to achieve suitable air removal.

Dynamic Vacuum. Once the maximum initial vacuum

has been reached, the vacuum pump is kept running
while a 5–10 min. steam-injection is performed from
the side of the chamber that lies opposite the vacuum
drain point. Modern autoclaves can perform both
methods depending on the load to be sterilized.

Heating and sterilization phases

Considerable amounts of condensate form in the

Fig. 3 Dovetail section gasket activated by compressed air. chamber during the heating and sterilization phases.
(From Ref.[5].) This condensate must be removed, and there are
 3534 Sterilization: Moist Heat
Sterilization
Statistical–

basically two ways to accomplish this. The first uses a Cooling with Cold Water Sprayed Directly onto the
condensate trap at the bottom of the chamber. This is Load with Air Counterpressure. Very frequently, the
the cheapest and simplest method, but it causes signifi- pressure stress that occurs when using method of
cant drops in pressure (and therefore in temperature) cooling by direct spraying of cold water, cannot be
when the trap opens, owing to the inertia of the trap. tolerated by the load. In such cases, it is possible to
The second method uses dynamic steam. This is the drain the steam from the chamber by replacing it with
most reliable method, but it is also slightly more sterile compressed air at a pressure equal to or greater
expensive. During the heating and sterilization phases, than the sterilization pressure. Cooling water is sprayed
the vacuum pump runs continuous and extracts the onto the load only after this replacement has been per-
condensate through a small valve. A small amount of formed. However, it is obvious that this method only
steam of course is also extracted continuously, accord- allows for reduction of the pressure stress of the con-
ingly providing the dynamic condition of the steam. tainers in the cooling phase; the pressure stress in the
sterilization phase (discussed later) is unavoidable.
Poststerilization phases
Ampoule Tightness Tests. The purpose of these tests is
These phases may be very different and are clearly to allow for rejection of ampoules that have closure
linked to the sterilized material and to the required defects, fractures, or cracks. These tests fall essentially
results. The most common solutions are the following: into two categories: penetration of dyed solutions
(usually with methylene blue) in the ampoules and post-
sterilization pressure stress. Details of these methods
Drying–Cooling Final Vacuum. This is produced by
are not presented here because of space constraints.
restarting the vacuum pump until a preset vacuum
(e.g., 100 mbar) is reached. The pump is then kept run-
Sterilization of the air that enters the chamber
ning for a preset time. Porous materials (and non-porous
materials also) are thus dried and cooled quickly.
As noted previously, in many cases it is necessary to
introduce air in the sterilization chamber. This air must
Cooling by Circulating Cold Water in the Jacket. This be sterile, otherwise it recontaminates the sterilized load
method is used with containers that are partially filled or the sterile room if a two-door autoclave is connected
with solutions (for example, culture media) and closed to it. This air is generally sterilized by filtration with a sys-
with non-hermetic closures. With such containers, tem built into the autoclave. Therefore, it is necessary to:
drying–cooling final vacuum is not applicable because
the solution would boil, and cooling by direct spray 1. Provide a filtration cartridge with suitable reten-
(described hereafter) may cause contamination. Steam tion.
is removed from the chamber by introducing sterile air 2. Allow in situ periodic sterilization of the
at a pressure that is equal to, or greater than the ster- assembled system by means of an automatic
ilization pressure. Then cold water is circulated in the process of the autoclave.
jacket. Chamber air pressurization has two purposes: 3. Ensure that the filtration system and its piping
to prevent boiling of the solutions and to improve heat maintain sterility during successive sterilization
exchanges between the load and the jacket. programs used for production.
4. Perform the system integrity test before and
Cooling by Direct Spraying of Cold Water onto the after each sterilization program of the filtration
Load. This method is generally used for cooling filled system.
and sealed ampoules contained in perforated trays and 5. Allow for validation of all procedures noted
generally marshaled. It is performed by spraying, or earlier.
rather by nebulizing, purified water or water for injec-
tion onto the load by means of a sparger located in the Process controllers
ceiling of the chamber. Water nebulization produces
rapid steam condensation and an equally rapid pres- Today, process controllers installed in autoclaves are
sure drop in the chamber while the pressure inside based on programmable logic controllers (PLCs),
the ampoules remains high (because the temperature personal computers (PCs), customized electronic solu-
of the solution decreases rather slowly). However, tions, or, sometimes, different combinations of the
good-quality ampoules can withstand this treatment aforementioned systems. However, a very large num-
adequately. The water spray is generally stopped when ber of autoclaves managed by old electropneumatic
the load temperature reaches 70–80
C. Accordingly, systems are still in operation. Modern process control-
the load still contains enough heat energy to dry lers, of course, offer previously inconceivable levels of
spontaneously once removed from the autoclave. performance. Today, temperature and/or pressure
Sterilization: Moist Heat 3535

Sterilization
Statistical–
control is generally performed with a proportional- Obviously, the overpressure depends on the filling tem-
integral-derivative algorithm. Steril-ization can be perature, the sterilization temperature, the ratio between
time-managed or F0-managed (F0 being accumulated solution volume and head volume, etc., but at 121
C, it is
by several flexible temperature probes enabled for this on average approximately 1.4 bar. Clearly, this phenom-
function). Some management systems offer exceptional enon cannot be ignored: suffice it to note that the stopper
flexibility in composing programs and setting parameters. of glass bottles with a mouth having a cross-section of
Information provided in real time on cathode ray tube approximately 4 cm2 would be subjected to an expulsion
(CRT) or liquid crystal display (LCD) or produced/ force of approximately 6 kg.
stored on paper/electronic media is highly detailed. These conditions therefore prohibit the use of tra-
ditional pure saturated steam autoclaves to sterilize solu-
tions contained in a wide variety of containers such as
DIFFERENTIAL PRESSURE BETWEEN INSIDE/
OUTSIDE OF A RIGID CONTAINER, PARTIALLY 1. Large-volume parenterals (LVPs) in glass bottles;
FILLED WITH WATER SOLUTION AND SEALED, 2. LVPs and small volume parenterals (SVPs) in
DURING STEAM STERILIZATION plastic containers (flexible, semirigid, rigid);
3. Prefilled glass or plastic syringes;
When a container in the conditions noted earlier is 4. Jars and similar containers with press-on or
sterilized in a conventional autoclave that operates screw caps;
with pure saturated steam, during sterilization a 5. Blisters containing various materials, for
considerable overpressure with respect to the pressure example, disposable contact lenses.
inside the autoclave chamber is generated in the con-
tainer. This is clearly attributable to the fact that the To correctly sterilize these products, it is necessary
air (or gas) that was present at filling has remained in or advisable to use a counterpressure autoclave.
the container, whereas the air was eliminated from
the autoclave chamber at the beginning of the process.
Fig. 4 schematically explains the phenomenon in ideal COUNTERPRESSURE MOIST-HEAT
conditions, i.e., considering air a perfect gas. STERILIZATION
Experimentally, it turns out that the actual overpres-
sure is higher than the theoretical one. This is attributable Moist-heat autoclaves operating with counterpressure
to various facts: the thermal expansion of water is greater are sterilizers capable of controlling the pressure of
than the thermal expansion of the glass of the container; their sterilizing medium independently of its tem-
the solution contains dissolved gases that come out of the perature. They are used essentially for the terminal
solution as the temperature rises; air is not a perfect gas. sterilization of solutions.

Fig. 4 Schematic description of pressures produced inside a rigid container, partially filled with water solution and sealed,
during steam sterilization at 121
C. (Adapted from Ref.[5].)
 3536 Sterilization: Moist Heat
Sterilization
Statistical–

Accordingly, a dual control principle is provided sterilization phase is generally very good: much better
that acts independently on both parameters. Two than 1.0
C. The cooling phase is performed by the
methods currently in use are superheated water spray same circulating water, which is now sterile and con-
and steam–air mixture. tinuously recirculated through the heat exchanger, in
which cold water (instead of steam) now flows without
contact with the sterile circulating water. The tempera-
SUPERHEATED WATER SPRAY AUTOCLAVES ture inside 500 ml containers drops to approximately
80
C in 10–12 min. This temperature is generally
Fig. 5 is a typical diagram of these autoclaves. Alterna- suitable to obtain rapid and spontaneous drying of
tives are possible, but they do not alter the essential the load once removed from the autoclave.
structure. The chamber is horizontal and generally An appropriate partial pressure of air (sterilized by
cylindrical, with a single wall and rectangular door(s) filtration) is maintained in the chamber during every
inscribed in the circumference. phase of the process, to compensate for the overpres-
At the beginning of the process, after loading the pro- sure inside the containers. Various methods for
duct, the lower part of the chamber is filled with water of controlling the total chamber pressure (steam þ air)
adequate chemical and bacteriological quality. The air can be used. With computerized process controllers, it
contained in the chamber is not removed. A sanitary- is also possible to correlate at any time during each
type pump circulates the filling water through a heat phase the air partial pressure to the average of the solu-
exchanger (of the removable-plate or other sanitary tion temperatures of two or more reference containers.
type) that is indirectly heated in countercurrent with Consequently, the load suffers no thermal or press-
plant steam. The water is then sprayed onto the load ure shocks because the differential pressure between
by a sparger located in the upper part of the chamber the chamber and the containers can be reduced to zero
and equipped with a system of solid-cone spray nozzles. or maintained at all times during the process in a direc-
Uniform water redistribution in the lower layers of the tion that is suitable for the particular type of container
load is ensured by suitable perforated racks that support during sterilization or, generally, during thermal treat-
the product. Sometimes additional water spray bars are ments from 60–127
C.
located on both sides of the chamber. Clearly, these autoclaves have some limitations in
Heating of the circulating water and, therefore, of their application:
the load is very gradual but quite rapid. A temperature
of 121
C is typically reached in 25–30 min inside 500 ml 1. It is impossible, or illogical, to dry the load
containers; the heating rate clearly depends on the inside the autoclave by pulling vacuum in the
characteristics of the solution and its containers. chamber or by circulating warm air through
Temperature uniformity in time and space during the the chamber and the load.

Fig. 5 Superheated water spray autoclave: simplified P.&I.D. (Adapted from Ref.[5].)
Sterilization: Moist Heat 3537

Sterilization
Statistical–
2. If materials that have upward-facing concave the chamber. This is an important and demanding task
surfaces are sterilized, these surfaces will be because the air clearly tends to stratify on the bottom.
filled with water at the end of the process. The The condensate that forms is removed by continuous
obvious remedy is to load the material upside and spontaneous bleeding from the chamber.
down. The cooling phase consists of feeding compressed
3. When sterilizing solutions contained in PVC and sterile air to the chamber to condense and replace
bags, so-called blushing can occur, i.e., the all the steam, while maintaining the same total sterili-
PVC can whiten because of water absorption. zation pressure or possibly increasing it. Cold plant
The time required for this blushing to disappear water is then fed to the internal heat exchangers, which
can be quite long, depending on the type of PVC are constituted by batteries of hollow plates arranged
and its plasticizer. Blushing does not occur with in the two lateral sectors of the chamber (for simplicity,
polypropylene (PP), polyethylene (PE), and only one plate is shown in Fig. 6). However, this cool-
polylaminated plastics. ing method uses two solid-gas heat exchanges, which
have poor efficiency. An attempt can be made to
These autoclaves are sterilizers that can vary consider- improve efficiency by increasing the air pressure in
able in size but are generally rather large (1–20 m3 and the chamber within the limits of the product, thus
more). They are often provided with automated loading/ increasing the density of the air and therefore its
unloading systems. exchange efficiency. The fans of course continue to
run during the cooling phase as well.
Despite this refinement, the cooling phase is signifi-
STEAM–AIR MIXTURE AUTOCLAVES cantly longer than that in superheated spray water
autoclaves. A mechanically critical point of steam–air
Fig. 6 is a typical diagram of these autoclaves; possible autoclaves is the tightness of the fan shaft. This prob-
alternatives are addressed later. The chamber is similar lem has been completely solved in the more advanced
to that of superheated water spray autoclaves. At the machines by adopting magnetically driven fans (Fig. 7).
beginning of the program, steam enters the chamber The air partial pressure during the program is mana-
directly through a suitable sparger located in the lower ged as described above for superheated water spray
part of the chamber. The air initially contained in the autoclaves, and the dimensions and loading/unloading
chamber is not removed. The high-efficiency fan(s) systems are also similar.
located on the ceiling of the chamber and the flow Possible alternatives to the configuration shown in
deflector system have the task of homogenizing and Fig. 6 are 1) horizontal fans (instead of vertical fans)
circulating the steam–air mixture that forms inside located on one side of the chamber. This solution

Fig. 6 Steam-air autoclave: simplified P.&I.D. (Adapted from Ref.[5].)

 3538 Sterilization: Moist Heat
Sterilization
Statistical–

Fig. 7 Schematic drawing of magnetically driven fan. (Adapted from Ref.[5].)

entails a more severe risk of shaft bending and the positive features of steam–air autoclaves is the rela-
vibration, which can cause wear of the delicate sealing tive ease in combining the traditional pure saturated
system. Moreover, it is technically more difficult to steam cycles, i.e., in manufacturing hybrid pure
manufacture magnetically driven fans with a horizontal steam/steam–air autoclaves (in this case, the chamber
shaft; and 2) shell-and-tube heat exchangers (instead of is equipped with a jacket and a vacuum pump). This
plate-type exchangers). combination is instead not recommended for super-
With steam–air autoclaves, blushing of PVC bags is heated water spray autoclaves, although it is offered
generally less intense than with superheated water by some manufacturers. Fig. 8 is a summary compa-
spray autoclaves and essentially affects only the areas rison of superheated water spray and steam–air
where the bags rest on the supporting racks. Among autoclaves.

STERILIZING A ROTATING LOAD

Currently, the pharmaceutical market more and more

often requires rotating-load sterilization autoclaves.
Load rotation can have essentially three goals:

1. To maintain the stability (or homogeneity) of

emulsions (or suspensions) that would tend
to break out because of the sterilization tem-
perature.
Fig. 8 Critical comparison of superheated water spray 2. To sterilize heat-sensitive products at high tem-
(SWS) and steam–air mixture (SAM) autoclaves. (Adapted perature, drastically reducing the sterilization
from Ref.[5].) time. The logic behind this principle is that, as
Sterilization: Moist Heat 3539

Sterilization
Statistical–
Fig. 9 Two superheated water spray autoclaves with a chamber capacity of approximately 4 m3, with a rotating load and sliding
double doors. The man–machine interface of the process controller is not shown. The automated loading/unloading systems,
frequently used with large sterilizers to allow faster loading and unloading operations, are shown instead. (Adapted from
Ref.[5].)

noted, the temperature coefficient of the moist- impossible to avoid mass displacements of the load
heat sterilization reaction is very high (z is on during rotation; and 4) lubrication of the load bearing
the average equal to 10), whereas the tempera- and rotating system must be avoided for hygiene-
ture coefficient of a classic thermal degradation related reasons.
reaction is much lower (on the average equal to Finally, it is evident that the actual loading capacity
2). Obviously, to achieve the goal, the product of the chamber is reduced because of the presence of
heating/cooling sterilization rates must be very a cylindrical structure that must rotate inside it and
high and uniform. Because rotation stirs the support the load contained in appropriate trays with
product, it indeed facilitates the penetration/ a lid (the entire system being appropriately perforated).
removal of heat into/from the product, espe- Fig. 9 shows this cylindrical structure both when empty
cially if it is dense and viscous. This principle and when filled with the trays. These autoclaves are
is currently often used to sterilize heat-sensitive generally counterpressure sterilizers and the load is
LVPs, for example, glucose solutions (especially rotated throughout the process at an adjustable rate
with a high concentration of sugar) or amino (1–10 rpm) and, if required, intermittently and in
acid solutions. alternating directions.
3. To provide the best possible testing of ampoule
tightness with fast poststerilization vacuum.
This testing method (presented above) achieves
maximum effectiveness when the ‘‘open’’ REFERENCES
defects of the ampoules are below the level of
the solution. Ampoule rotation is the ideal 1. Carleton, F.J.; Agalloco, J.P. Validation of Aseptic Pharma-
method for achieving this condition regardless ceutical Processes; Marcel Dekker, Inc.: New York, 1986.
2. Lavagna, S.M. Injectables and Water for Pharmaceutical
of the location of the defect on the ampoule Use; Editrice Bias: Milan, 1995.
(tip, shoulder, bottom). 3. Pflug, I.J. Syllabus for an Introductory Course in the Micro-
biology and Engineering of Sterilization Processes, 4th Ed.;
Environmental Sterilization Service: St. Paul, MN, 1980.
Naturally, the production of this type of autoclave 4. Validation of Steam Sterilization Cycles; Technical Mono-
requires highly refined design and construction tech- graph No. 1; Parenteral Drug Association: Philadelphia,
nology because 1) the load rotation system complicates 1978.
5. Pistolesi, D.; Mascherpa, V. Moist and Dry Heat Steriliza-
construction significantly; 2) the loads to be rotated tion Technology; Fedegari Autoclavi S.p.A.: Albuzzano,
are generally bulky and heavy; 3) it is practically PV, 1999; CD-ROM.
Sterilization
Statistical–

Sterilization: Radiation
Stephen G. Schulman
Department of Medicinal Chemistry, University of Florida,
Gainesville, Florida, U.S.A.

Phillip M. Achey
Department of Microbiology and Cell Science, Institute of Food and Agricultural Sciences,
University of Florida, Gainesville, Florida, U.S.A.

INTRODUCTION exposure of a human to an amount of ionizing radi-

ation energy equivalent to the amount of thermal
The actions of ionizing radiations on matter and the energy received by drinking a cup of hot coffee will
subsequent interactions of the irradiated molecules result in the death of the individual within 30 days.
are useful for the sterilization of pharmaceutical and Therefore, because of the potent biological action
surgical supplies. To take maximum advantage of the and the lack of our ability to sense exposure to ionizing
benefits derived from ionizing radiations, it is desirable radiation (unless the exposure dose rare is extremely
to have an understanding of the fundamental processes high), it is important to practice extreme caution when
that result in radiation damage to living and non-living working with this agent.
systems. Another common characteristic of ionizing radi-
This article addresses with the effects of ionizing ation is its ability to penetrate material. The two most
radiation on biological systems, beginning with the commonly used forms for sterilization are energetic
physical and chemical actions on condensed matter. electron beams and electromagnetic radiation (e.g.,
After these sections, the biological responses of living g-rays from cobalt-60 or cesium-137). The penetrating
systems, particularly microorganisms and viruses, to ability of g-rays and X-rays is much greater than that
ionizing radiation are considered. A section on the cur- of electrons. In either case, when developing protocols
rent uses and government policies on the applications for using ionizing radiation insterilization of material, the
of ionizing radiation in areas important to the pharma- penetrating ability of the radiation must be considered.
ceutical industry is also included.

PHYSICAL AND CHEMICAL ACTIONS

Description of Ionizing Radiation OF IONIZING RADIATION

Radiation energy can be either in the form of electro- This section deals with the fundamental nature of the
magnetic energy or of particle radiation. Radiation is interactions of high-energy radiations with matter,
distinguished as being either non-ionizing or ionizing. from the absorption of the radiations to the eventual
Examples of non-ionizing radiation include the ultra- establishment ofchemical equilibrium in the system.
violet, visible, infrared, and radiofrequency parts of The process may bedivided into three stages which
the electromagnetic spectrum. These kinds of radiation are illustrated in Fig. 1.
are not addressed in this article.
A common feature of all ionizing radiation is that it 1. The physical stage, consisting of the absorption
is of sufficient energy to cause ionizations in the of the radiant energy by the irradiated system.
exposed material. These ionizations result in release Its duration is of the order of 10
15 s.
of orbital electrons from atoms and cause disruptions 2. The physicochemical stage, the processes that
of covalent bonds. Release of energy acquired by mole- lead to the establishment of thermal equilibrium
cules in this manner will result in dramatic changes in in the system. Its duration is of the order of 10
12 s.
the physical and chemical structures of the exposed 3. This chemical stage, which entails diffusion and
materials because of the concentration and localized chemical reaction of the reactive species, ulti-
release of the ionizing radiation energy. By compari- mately resulting in chemical equilibrium. Its
son, thermal heating is less efficient at causing bond lasts upwards of 10
8 s, depending on the rate
rupture because of the wide distribution and diffuse constants and diffusion coefficients of the
release of thermal energy in matter. Whole-body reactive species.
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-100000439
3540 Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved.
Sterilization: Radiation 3541

Sterilization
Statistical–
Incident and I is a mean excitation potential for the medium
Time Incident charged X-ray or
scale particle gamma ray (I ¼ 11.5 Zev for Z 30, I ¼ 8.8Z ev for Z > 30,
photon where Z is the mean atomic number of the medium).
The term
dE/dx is called the linear energy transfer
10-18
to
(LET) of the radiation. If electrons (b-rays) are the
Fast Physical
10-15 electrons stage ionizing particles, the expression for LET is slightly
sec. different:
Ionizations
 2 rffiffiffi

dE 4pZ2 e4 n mv e
10-15

2
1n ð2Þ
Physico
dx mv 2I 2
to
10-12 chemical
sec. stage where e is the basic of the natural logarithms.
Free
radicals Several important conclusions can be drawn from
Eqs.1 and 2. First, the rate of energy loss of a charged
10-12
particle in a given medium is proportional to the elec-
to Chemical tron in the medium. Second, because the factor v2 out-
10-3 Chemical stage side the logarithmic term is more important than that
sec. changes inside, the rate of energy loss increases as the particle
slows down. Third, if two particles of equal energy
but different mass are compared, the heavier one
Seconds will have a smaller velocity and thus a higher LET.
to Biological
years
Consequently an a-particle will produce many more
stage
Biological excitations and ionizations per unit path length and
changes have a shorter path length than a b-particle of the same
(death or
mutation) energy.
Fig. 1 The stages of radiation action on chemical systems
and biological organisms. High-energy photons

When high-energy photons such as x-rays and g-rays

The Physical Stage pass through matter, they lose energy by way of three
mechanisms: photoelectric absorption, in which the
The absorption of the energy associated with ionizing photon transfers its entire energy to an electron;
radiation by a medium results, initially, in ionization Compton scattering, in which the photon transfers part
and electronic excitation. These processes occur of its energy to an electron; and pair production, in
regardless of the nature of the radiation. The mech- which the photon disappears and a high-energy elec-
anism of excitation and ionization by charged particles tron and positron are formed. The relative importance
is different, however, from that effected by high-energy of each of these mechanisms depends on the energy of
photons. the photon. For photons in the 100-kev to 2-MeV
range, the principal mode of absorption by the medium
Charged particles is Compton scattering. Much higher photon energies
favor pair production (at least 1.02 MeV are required
The interactions with a medium of charged particulate to produce a pair), whereas lower energies favor photo-
radiations such as protons, (b-particles, and g-rays electric absorption. The principal effect of the absorp-
consist predominately of electrostatic coulomb exci- tion of high-energy photons is the production of
tation and ionization caused by ejection of atomic energetic electrons that then dissipate their energies
and molecular electrons in the medium. According to by the mechanism described by Eq. (2).
Bethe’s semiclassic treatment, the energy lost to the In general, the effect of transfer of energy from an
medium, per unit length of path, by a heavy particle energetic particle to the medium is to produce along
of charge Ze and velocity v is: its path a variety of electronically excited molecules,
ions, and free electrons. Secondary electrons are also

dE 4pZ2 e4 n 2mv2 formed along with the ions. The electronic transitions

1n ð1Þ resulting in the formation of these species occur in
dx mv2 I
times [10
15 s] that are short compared with molecular
where n is the electron density (number of electrons per vibration periods [10
14–10
12 s]. The amount of
unit volume) of the medium, m is the electronic mass, energy absorbed by the irradiated system per unit mass
 3542 Sterilization: Radiation
Sterilization
Statistical–

is called the dose and is expressed in Rads, where and react with molecules in the bulk of the liquid
1 Rad (¼6.24  1013 eV/gm) is the amount of radi- medium. Those that recombine within the spur react
ation that will deposit 100 ergs of energy per gram of so rapidly that they cannot be detected by physical
the irradiated system. or chemical methods. They form stable molecular pro-
ducts, which are known as the molecular yield. Those
radicals that diffuse away from the spur and react with
The Physiochemical Stage the medium can be detected by physical methods such
as electron spin resonance spectroscopy and by chemi-
This stage lasts about 10
14–10
12 s, which is typical of cal methods such as compound formation with radical
the period of molecular vibrations. During this period, scavengers, e.g., iodine and diphenylpicrylhydrazyl.
internal molecular rearrangements can take place. The compounds formed by reaction with radical sca-
During the physiochemical stage, the excited mole- vengers are called the radical yield. The mechanisms
cules and ions dissipate their excess energy by bond of chemical radiation effects are frequently determined
rupture, luminescence, internal conversion, and energy by comparison of relative molecular and radical yields.
transfer to neighboring molecules. Also during this The radiation chemistry of liquids has developed along
stage, the low-energy, secondary electrons produced two distinct paths—that of water and aqueous
during the physical stage interact with molecules in solutions and that of organic liquids. The radiation
the environment resulting in the formation of free chemical processes responsible for the destruction of
radicals. microorganisms are probably most closely related to
the former, but damage to membranes and cell walls
The Chemical Stage may actually be better related to the radiation chemis-
try of organic liquids or even some solids.
During this stage, the reactive intermediates (ions and
radicals) produced in the previous stages diffuse away
from their sites of production and undergo chemical Water and Aqueous Solution
reactions with each other and with other molecules in
the environment. The irradiation of pure water is believed to result in
In condensed systems (liquids and solids), the main two dissociative processes. The first of these is the
reactive species produced in the physiochemical stage direct dissociation of water into hydrogen atoms and
that react in the chemical stage are free radicals. Their hydroxyl radicals:
primary modes of reaction are atomic abstraction, rad-
ical recombination, and addition to p-bonds. H2 O ! H þ OH ð3Þ
Because most systems of interest to pharmaceutical
scientists are either liquids or solids, it is useful to The second reaction is the ionization of water to
present some of the features of radiolysis common to yield a hydrogen ion, a hydroxyl radical, and a
liquid or solid samples. hydrated electron:

H2 O ! Hþ þ OH þ e

aq ð4Þ
Liquid
The hydrated electron is a powerful reducing agent
The irradiation of liquids results initially in the ejection and will reduce water and the hydrogen ion according
of electrons with the consequent formation of ions. to:
The ejected electrons usually lose their excess kinetic
energy within the electric field of the parent ion. Most

e

aq þ H2 O ! H þ OH
 ð5Þ
of the ion pairs formed culminate in recapture of the
ejected electrons, leaving the molecules in a highly and the hydrogen ion according to:
excited, electronic state that may return to the ground
state by internal conversion, luminescence, or energy e

aq þ H
þ
! H ð6Þ
transfer. Alternatively, the highly excited neutral mole-
cules may split into free radicals. Some ion pairs may Because the latter two reactions result in the same
be sufficiently long-lived to diffuse away from the site products as the direct radiolysis and because the pro-
of production and react with the surrounding medium. ducts of reduction by the hydrogen atom and the
The free radicals are the most important reactive spe- hydrated electron are identical, it is frequently impos-
cies formed. Once free radicals are formed along the sible to determine whether the hydrated electron or
track of an ionizing particle, they may combine with the hydrogen atom is the principal reducing species
each other or they may diffuse away from the spur in aqueous solutions. In acid solutions, it is reasonable
Sterilization: Radiation 3543

Sterilization
Statistical–
to assume that the hydrated electron will reduce Hþ (HO2.). This species acts principally as an oxidizing
almost exclusively and that H will therefore be the pre- agent; e.g.:
dominant reducing species. However, in neutral and
HO2 þ Feþ2 ! Feþ3 þ OH2


basic solution, the hydrated electron may be assumed ð10Þ
to predominate. The ultimate molecular products of
the radiolysis of pure water are hydrogen gas and An important consequence of this is that, whereas
hydrogen peroxide, formed by the reactions: solutions in pure irradiated water have approximately
equal oxiding and reducing capabilities, the presence
of oxygen in these solutions can result, in some cases,
H þ H ! H2 ð7Þ
in very strong oxiding properties because of the con-
version of the reducing hydrogen atom to the predomi-
OH þ OH ! H2 O2 ð8Þ nantly oxidizing hydroperoxy radical. In general, the
presence of oxygen in aqueous solutions will lead to
The radiation chemistry of aqueous solutions may alternations of the mechanisms of radiolyses owing
be considered from two points of view. The first, called to the ‘‘exclusively of oxidation.’’ Radiation damage
the Target Theory, considers the direct effect of ioniz- to microorganisms tends to be far more extensive in
ing radiations on the solute molecules. The second the presence of oxygen than in its absence.
approach regards transformations in the solute mole-
cules to be attributed to interactions with the reactive
intermediates formed by the radiolysis of water. Organic Liquids
Because most aqueous systems are relatively dilute,
the latter approach seems statistically more reasonable. An important difference between the radiation chemis-
Kinetic studies of dilute aqueous systems have indeed try of water and of organic liquids is that the concept
borne out this supposition. The radiation chemistry of the spur (a reasonably well-defined volume in which
of aqueous solutions then becomes the free radical the formation of the reactive species occurs along the
and redox chemistry of H, OH, and e
aq.
track of the ionizing particle) becomes hazy. The radi-
The effectiveness of radicals in producing chemical cals formed in water tend to recombine rather than
change in aqueous systems depends on the LET of react with the environment immediately after forma-
the ionizing radiation that produces these radicals. tion. The volume in which recombination is likely
A high LET particle, such as an a-particle or a proton, defines the spur. The radical products of irradiated
will produce a large concentration of radicals along its organic liquids, however, are more likely to interact
short track. These radicals are likely to recombine, with their immediate environment than to undergo
forming molecular products, before they can diffuse recombination. This is evidenced by the low molecular
away from the spurs in which they are formed. Low yields of hydrogen from irradiated organic systems.
LET particles, on the other hand, produce low radical The radiation chemistry of hydrocarbons and their
concentration along their tracks. This minimizes the derivatives has been investigated extensively. An
probability of recombination so that the radicals can important difference between gas phase and liquid
diffuse away from the spurs and initiate chemical reac- phase radiolysis of hydrocarbons exists in that the
tions. Protons and a-particles therefore result in high breaking of carbon–carbon bonds is an important pri-
molecular yields, whereas b- and g-rays result in high mary process in the gas phase, whereas in the liquid
radical yields. phase, the rupture of carbon–hydrogen bonds is almost
One of the earliest devices for the measurement of exclusive. Another important difference between anal-
radiation dosage, the Fricke dosimeter, is based on ogous reactions in gas and liquid phases occurs in the
the oxidation of the ferrous ion by OH radicals pro- polymerization process. In gas phase polymerizations,
duced in the radiolysis of a dilute aqueous solution the presence of radical scavengers such as iodine and
of ferrous sulfate: benzoquinone does not appreciably alter the yields of
polymeric products. In the liquid phase, however, the
yields of the polymers obtained from the irradiation
Feþ2 þ OH ! Feþ3 þ OH
ð9Þ of materials such as vinyl chloride are seriously cur-
tailed by the addition of radical scavengers. This indi-
The presence of dissolved oxygen alters the nature cates that polymerization in the liquid state occurs
of the redox properties of irradiated water. This is a primarily by a free radical mechanism, whereas in the
consequence of the ‘‘radical scavenging’’ property of gaseous state, it occurs by an ionic mechanism. The
oxygen. Molecular oxygen has two unpaired electrons. irradiation of polymeric materials results in cross-
One of these can form a covalent bond with a linking of polymer chains and grafting of dissimilar poly-
hydrogen atom, forming the hydroperoxy radical meric materials. This treatment of polymers contributes
 3544 Sterilization: Radiation
Sterilization
Statistical–

considerable tensile strength and heat resistance to chemistry of other aromatic hydrocarbons. The resist-
the irradiated polymers and is already being exploited ance of polystyrene (-(C6H5) CH-CH2-)n to cross-linking
commercially in the production of stain-resistant textiles compared with polyethylene is further evidence of the
and heat-resistant plastic containers. stability of aromatics to radiation effects.
Irradiation of saturated aliphatic compounds typi- Aromatic compounds frequently protect other more
cally results in unsaturation, polymerization, and iso- radiosensitive compounds from radiolysis. For
merization. The radiolysis of cyclohexane illustrates example, liquid cyclohexane is protected from exten-
all three of these processes. If the radicals are very sive radiolysis by the addition of a small amount of
energetic, cyclohexene can be formed by the abstrac- benzene. This is probably due to energy transfer from
tion of hydrogen from a cyclohexyl radical either by cyclohexane to benzene, followed by dissipation of the
a hydrogen atom or by another cyclohexyl radical. If excitation energy by the aromatic p-system.
the radicals become thermalized, recombination of One of the most important general features of the
radicals can occur to give bicyclohexyl. A less frequent radiation chemistry of liquids is that so much energy
process is rearrangement, followed by hydrogen atom is deposited by the ionizing radiations, excited or
capture to yield methylcyclopentane. reactive molecules are formed in close proximity and
The irradiation of alkyl halides results in cleavage of are likely to react with one another. This situation is
the carbon–halogen bond. The radiolysis of methyl not encountered inphotochemistry except when lasers
iodide, for example, yields ethane and molecular iodine. are used for excitation.
Alcohols, on radiolysis in the liquid state, yield alde-
hydes and vicinal glycols. For example, consider the
radiolysis of methanol: SOLIDS

CH3 OH ! CH2 OH þ H ð11Þ Pharmaceutical and surgical supplies are often in the
solid state when irradiated. Certainly, their containers
 CH2 OH ! CH2 ¼ O þ H ð12Þ are solid. It is therefore in order to consider some of
the radiation chemistry of solids.
and Because of the ‘‘fixed’’ positions of atoms in crystal-
line lattices, the effects of irradiation of solids include
2CH2 OH ! CH2 OHCH2 OH ð13Þ atomic displacements as well as electronic excitation
and ionization. Although electronic alterations of
The irradiation of frozen alcohols results in deep materials affect their chemical behavior, atomic displa-
coloration of the alcoholic glasses. Methanol turns a cements in solids are found to have a much more pro-
brilliant purple, whereas ethanol turns blue. These nounced effect on the physical properties of crystals.
colored glasses are stable if kept in the dark at low tem- To dislodge an atom from its normal lattice position,
perature. Exposure to visible or ultraviolet light results a certain amount of energy must be transferred to
in bleaching of the alcoholic glasses as well as in the the atom by an irradiating particle. Because of the
elimination of the electron spin resonance signal large mass of the atoms, electrons and photons will
observed in the colored glasses. The colors are believed be relatively ineffective in producing substantial
to be caused by the absorption spectra of trapped free numbers of atomic dislocations. The heavier particles,
radicals in the glasses. The product yields from the a-particles, protons, deuterons, and neutrons, will be
bleached glasses are different from those of irradiated much more effective at this process. Furthermore,
glasses that have not been exposed to light. This sug- unlike the primary effect of ionizing radiations in pro-
gests that the trapped radicals might be photolyzed ducing distal electronic disturbances through electro-
by visible and ultraviolet light. static effects, the predominant process that is required
The irradiation of aromatic compounds results in to produce atomic dislocations is direct collision.
considerably lower yields of radiolysis products than There are two types of lattice defects that occur in
does irradiation of aliphatic compounds of similar molec- all real crystals and at very high concentration in irra-
ular weight and functional group composition. This diated crystals. These are known as point defects and
has been attributed to effectiveness of the delocalized line defects. Point defects occur as the result of displa-
p-orbitals in accommodating excitation energy without cements of atoms from their normal lattice sites. The
permitting the molecule to dissociate. Nevertheless, some displaced atoms usually occupy sites that are not in
radiolysis does occur. Benzene is known to yield biphe- the lattice framework; they are then known as ‘‘inter-
nyl, phenylcyclohexadiene, and a polymeric material of stitials.’’ The empty lattice site left behind by the inter-
average composition (C6H7)x, which behaves as if it were stitial is called a vacancy. Avacancy produced by
an unsaturated hydrocarbon. Dimerization and polymer displacement of an anion or cation, along with its
formation are also characteristic of the radiation interstitial ion, is called a Frenkel pair, or simply a
Sterilization: Radiation 3545

Sterilization
Statistical–
Frenkel defect. In some cases, the displaced ions are Valence Crystals
removed so far from their vacancies that they form a
new layer at the crystal surface. The vacancies left The strong bonding in valence crystals results in the
behind in this case are called Schottky defects. Frenkel failure of these crystals to demonstrate, on irradiation,
and Schottky defects play very important roles in the quasichemical changes such as depolymerization.
properties of solids altered by radiation damage. Unlike metals, however, valence crystals have no
Line defects (dislocations) are produced by slippage conduction electrons. This permits them to retain
or shear of the crystal lattice. If the slippage is perpen- electronic dislocations as well as atomic displacement.
dicular to a face of the crystal so that the lattice planes The trapping of dislocated electrons in the crystal by
on either side of the dislocation are parallel but dis- potential wells such as those created by atomic vacan-
placed with respect to one another, the defect is called cies results in coloration of the normally transparent
an edge dislocation. If the slippage is angular, as if pro- valence crystals.
duced by rotation about the shear axis so that lattice
planes on either side of the defect are not perpendicu-
lar, the defect is called a screw dislocation. Ionic Crystals
There are four broad classifications of crystal types,
according to the nature of the interatomic forces hold- Irradiation of ionic crystals results in atomic and elec-
ing the crystal together. In metallic crystals, the atoms tronic dislocations. The trapping of displaced electrons
are thought to form a quasi-ionic lattice arrangement by anion vacancies results in the absorption of visible
with the valence electrons, which bind the lattice, delo- and near ultraviolet light, which give these crystals
calized throughout the crystal so that they cannot be their characteristic colors. These pseudoatomic elec-
identified with any atom. Valence crystals, such as dia- trons and their vacancies are called color centers.
mond, consist of a lattice in which the atoms are The exposure of colored ionic crystals to visible or
bonded by conventional covalent interaction through- ultraviolet light causes the annealing of trapped elec-
out the lattice. This implies that a valence crystal could trons and results in bleaching of the colorations
be considered a giant molecule. Molecular crystals induced by irradiation. In some cases in which the crys-
(e.g., naphthalene and water) are regular arrangements tal remains uncolored upon irradiation, thermolumi-
of well-defined molecules that are bound together in nescence is observed in the annealing process.
the lattice by Van der Waals and hydrogen-bonding The dissolution of a heavily irradiated crystal of
forces. Finally, the ultimate in electronic localization sodium chloride in water will result in the evolution
occurs in ionic crystals, in which the lattice is com- of hydrogen and chloride from the solution. The solu-
posed of alternating positive and negative ions held tion also turns alkaline. This is presumably owing
together by strong electrostatic attractions. Sodium to the reactions of trapped holes and electrons with
chloride is a typical example of an ionic lattice. water:

1
e
þ H2 O ! H2 þ OH
ð14Þ
Metallic Crystals 2
1
Because of the delocalization of electrons throughout holeþ þ CI
! C12 ð15Þ
the metallic crystal, no persistence of ionization or 2
chemical decomposition can occur because a positive Trapped electrons also account for the ability of
hole formed by an electron ejection is always refilled by irradiated sodium chloride to initiate polymerization
an electron from the conduction band. On the other hand, in acrylonitrile.
sufficiently energetic radiations can cause atomic displace- Irradiation of nitrates, chlorates, perchlorates, and
ments. The production of interstitial atoms swells the lat- bromates results in the liberation of oxygen. In KClO4,
tice, thereby decreasing the density of the crystal. irradiation results in explosion of the crystal due to the
The most obvious evidence of radiation damage in internal buildup of oxygen.
metallic crystals is decrease in electrical and thermal
conductivity. This is attributable to scattering of elec-
trons and phonons by vacancies and interstitials that Molecular Crystals
destroy the order of the lattice necessary for high
conductivity. The irradiation of substances that form crystals con-
The obvious effects of radiation damage in metallic taining discrete molecules held together by dispersion
crystals can be reversed by ‘‘annealing.’’ Heating the forces results in radiolysis in the conventional sense.
irradiated materials supplies the energy required to For example, the radiolysis of aliphatic carboxylic acids
push an interstitial back into a vacancy. in the solid state yields hydrogen, carbon monoxide,
 3546 Sterilization: Radiation
Sterilization
Statistical–

and carbon dioxide. The relative yields of these gases in a particular bond usually requires 10
13–10
9 s. To
depend on the strengths of the bonds involved in radi- remove energy or charge from an activated molecule
olysis and their frequency of occurrence. These consid- effectively, the protector should have a slightly lower
erations apply as well to liquids and gases and suggest ionization or excitation potential. In fluid systems,
no special solid-state effects. the rate of charge transfer can be limited by diffusion,
Energy transfer in molecular crystals seems to be a in which case, the donor and acceptor must be in con-
well-established phenomenon. Irradiated crystals of tact. Excitation energy, however, can be exchanged
anthracene containing only a trace of naphthacene radiationlessly by molecules as much as 70 Å apart
show the characteristic green fluorescence of naphtha- by reasonance energy transfer, a dipole–dipole inter-
cene rather than the violet of the primary constituent. action. The process is more rapid than fluorescence
If the material is dissolved in benzene, the anthracene and competes favorably with dissociation. Transfer
fluorescence predominates. processes of the resonance type are extremely efficient
The irradiation of ice results in formation of in crystalline materials in which the high degree of order
trapped hydrogen and hydroxyl radicals as well as permits excitation energy to travel in excitons that trans-
the hydrated electron. verse the crystal in a time shorter than its vibrational
The irradiation of surface catalysts alters the relaxation time. Crystalline structure also facilitates
properties of these catalysts through defect production charge transfer by providing conduction bands in which
on their surface. These defects have been observed to electrons can freely move about.
enhance and inhibit catalytic activity in specific cases. Energy conversions within a molecule can decrease
For example, the irradiation of silica gel enhances the the probability of decomposition; energy can be dis-
rate of H2–D2 exchange on it. tributed so widely that its localization in any one bond
is improbable. Aromatic compounds are protective
because they can probably dissipate acquired exci-
CHEMICAL PROTECTION FROM tation energy throughout their extensively delocalized
IONIZING RADIATIONS p-systems.

The effects of ionizing radiations on chemical and bio- Scavenging Intermediates

logical systems may be minimized by the addition of
certain chemical compounds to the system to be The addition of certain compounds that react readily
irradiated. These compounds react either directly with free radicals can effectively prevent these products
with the radiation or, more often, with the reactive spe- of ionizing radiations, from causing secondary damage
cies produced by the radiations. In so doing, they are in the system. Molecular iodine is a very effective
themselves transformed into other substances, but their radical scavenger forming iodocompounds with radi-
transformation results in the preservation of the integ- cals and leaving behind iodine atoms to do further
rity of the original chemical or biological system. scavenging, e.g.:
At the molecular level, there are several mechanisms
that account for the protection of irradiated systems CH3 þ I2 ! CH3 I þ I 
by chemical agents. These are energy and charge trans-
fer, in which an ionized or excited molecule transfers CH3 þ I3 ! CH3 I þ I 
its charge or excess energy to a protecting molecule
either by collision or by resonance transfer; scaveng- Oxygen is a diradical that enhances radiation damage
ing, in which a protecting radical scavenger reacts with by forming radicals with other radicals. An example
radicals from the initial actions of the radiation before is the scavenging of hydrogen atoms by molecular oxy-
they can attack other molecules in the system; and complex gen to form the hydroperoxy radical.
formation, in which a protective molecule can form com-
plexes that are either more or less susceptible to radiation
damage than the original substance. Complexes

Certain compounds may exert protective action by

Energy and Charge Transfer forming molecular complexes with the original mole-
cules of the system. These complexes might be less
The transfer of charge or excitation energy must be fast sensitive to radiolysis or attack by radicals, or they
enough to compete with dissociation processes if may be better able to transfer charge and excitation
protection is to occur. In some cases, an activated energy than the original compound. For example, the
molecule can dissociate within 10
14 s, the time for one radiolytic degradation of polyisobutylene is reduced
molecular vibration. However, localization of energy by copolymerization with styrene. The radiation
Sterilization: Radiation 3547

Sterilization
Statistical–
resistance of the porphyrin ring is enhanced by com- groups of carbohydrates are especially radiosensitive.
plexing it with vanadium and other metals. Mannitol is readily oxidized to mannose and sorbitol
The most obvious application of chemical protec- to glucose. Although oxidation of primary alcohol
tion from ionizing radiations is to biological systems. groups is favored by aerobic conditions, high yields
For a protective agent to be biologically practical, it from the oxidation of secondary alcohol groups are
must be non-toxic at protective concentrations, widely favored by anoxic conditions.
distributed, and remain intact for long periods before The irradiation of polysaccharides results predomi-
irradiation. Many substances have been applied to nately in their degradation. This explains why fruits
this problem. To date, the most effective have been and vegetables become soft on irradiation. This degra-
compounds such as cysteine because of the scavenging dation is observed to occur both in solution and in the
property of the –SH group and the ease of oxidation of dry state.
the –NH2 group.
Adventitious impurities in pharmaceutical and
Amino Acids and Peptides
surgical supplies may act as energy transfer acceptors,
scavengers, or complexants and make radiation sterili-
The irradiation of amino acids results in transforma-
zation more difficult owing to radioprotective action.
tion of both the amino and the carboxylic functions.
In the dry state, glycine is decarboxylated to methyla-
mine on irrradiation; in dilute aqueous solution, how-
MOLECULES OF BIOLOGICAL SIGNIFICANCE ever, the amino group is hydrolyzed to give glyoxylic
acid, acetic acid, and formaldehyde. In solutions of
There are two distinct theories of the actions of ioniz- concentration greater than 2%, methylamine again
ing radiations on the compounds of living cells that becomes an important product. The other amino acids
result in chemical transformation that leads to are similar to glycine in their radiolytic behavior. Ala-
mutation or cell death. The first is the target theory. nine, for example, gives ethylamine and CO2 in the dry
This approach regards only those events that produce state and pyruvic acid and ammonia in aqueous
direct ionizations in biologically significant molecules solution.
as being important. The evidence for this is that in The aromatic amino acids, when irradiated in aque-
many cases, the amount of damage to a given organism ous solution, show effects that are typical of aromatic
varies logarithmically with the dose of radiation. This compounds and amino acids. Phenylalanine is deami-
implies that the amount of damage possible in a cell nated in aerated solutions with the formation of a
is proportional to the number of radiosensitive mole- ketone. The aromatic ring remains relatively stable to
cules remaining undamaged and, therefore, capable radiolytic decomposition.
of reacting. The other theory is based on an indirect The irradiation of peptides results in a chemistry
relationship between the incident radiation and the similar to that of the amino acids but also in the break-
affected, biologically significant molecules. In this age of the peptide bond. In aqueous solution, all irra-
approach, the solvent, water, interacts with the radi- diated peptides give ammonia whether or not free
ation, forming ions and radicals. These reactive species, amino groups are present.
in turn, react with the biologically significant molecules The thiol and disulfide containing amino acids
causing radiation damage. Radiation biology, under this degrade to keto acids with the evolution of CO2 and
approach, is simply a branch of the radiation chemistry H2S, when irradiated in the dry state, In solution, how-
of aqueous solutions. There is evidence that both the tar- ever, the thiol and disulfide groups are excellent radical
get and indirect processes occur. scavengers, and free radical attack on these groups pre-
In this section, the effects of ionizing radiations on cludes deamination. The ultimate result of irradiation
molecules known to have biological significance and of thiol-containing amino acids is their oxidation to
the relationship of the radiation chemistry of these disulfides. Thus, irradiation of an aqueous solution of
molecules to radiation effects observed in living organ- cysteine results in the formation of cystine. The
ism are considered. irradiation of the disulfides results in higher oxidation
products. For example, cystine gives cystine disulfox-
ide in aqueous solution. Reduction of disulfides to
Carbohydrates
thiols is not observed.
The irradiation of aqueous solutions of carbohydrates
has the same effect as it does on alcohols. The hydroxyl Proteins and Enzymes
groups are attacked to yield carbonyl compounds.
Under anoxic conditions, dimer products and, ulti- The irradiation of proteins results in the formation of
mately, polymers are also formed. The primary alcohol free radicals at the sites of —S—S— bonds. Aromatic
 3548 Sterilization: Radiation
Sterilization
Statistical–

amino acids in the proteins are also particularly sus- Para-aminobenzoic acid is destroyed on irradiation
ceptible to radiolysis; decarboxylation and deamina- in aqueous solution by deamination and decarboxyla-
tion being common results of irradiation. Rupture of tion. Sulfanilamide and sulfathiazole are inactivated
the peptide linkage is characteristic of the radiolysis presumably because of deamination. The cobalt in
of proteins. In the case of enzymes, the destruction vitamin B12 is reduced on irradiation from þ3 to the
of peptide linkages is accompanied by a decrease in þ2 state.
biological activity. This decrease continues after The plant hormone auxin has been shown to be
irradiation is stopped. The mechanisms of radiolysis radiosensitive. The product of the irradiation of auxin
in the dry state and in solution are different, but the (b-indoleacetic acid) is a polymer similar to that
results are usually similar. One of the more important obtained in the radiolysis of indole.
differences between these results is the degradation of
proteins by dry-state irradiation compared with
increase of molecular weight through cross-linking in Nucleic Acids
solution. In general, the radiation chemistry of proteins
and enzymes may be considered a special case of the The nucleic acids DNA and RNA are responsible for
radiation chemistry of peptides and amino acids. the transmission of genetic information and protein
synthesis. Both of these processes are dependent
on the ordering of purine and pyrimidine bases
Respiratory Proteins, Vitamins, that are bound to the main body of the molecule by
and Coenzymes phosphoric ester linkages. The main body of these
molecules consists of ribose (5-carbon sugar) mole-
Respiratory proteins cules linked together by phosphoric acid units to form
a long strand. The purine and pyrimidine bases
These substances are iron, porphyrin, protein com- branch off from the chain at the ribose sites. The
plexes. Irradiation of these substances may produce DNA molecule consists of two helically intertwined
effects in the porphyrin ring or in the protein, but oxi- strands of nucleic acid held together by hydrogen
dation or reduction of the iron is almost always bonding between purine pyrimidine pairs on opposite
involved. The iron in ferricytochrome-c is reduced to strands.
the ferrous state under neutral conditions. Under acid The irradiation of nucleic acids ruptures hydrogen
conditions, the ferric form is favored. Hemoglobin and bonds that hold DNA strands together results in dea-
oxhemoglobin are both oxidized from the ferrous to mination and dehydroxylation of purine and pyrim-
the ferric state, destroying the property of oxygen idine bases, fission of sugar base linkage, liberation
transport. Large radiation doses result in attack on of the purine bases, destruction of the pyrimidine
the porphyrin ring and denaturation of the protein. bases, oxidation of the sugar moiety, and breakage of
When irradiated in the dry state in the absence of the nucleotide chain with liberation of inorganic phos-
oxygen, hemoglobin is not oxidized, but it becomes phates. In the presence of oxygen, irradiation leads to
insoluble because of protein denaturation. Myoglobin the formation of hydroperoxides of the pyrimidine
behaves in a manner similar to that of hemoglobin bases but not of the purine bases.
but is considerably more radiosensitive. Myoglobin is In general, the purine bases appear to be much more
a copper-containing respiratory protein of molecular stable to radiolysis then pyrimidine bases. This is prob-
weight more than 10 times that of hemoglobin. In ably owing to the greater p-delocalization energy of the
this case, attack at the protein part of the molecule purines, which provides a pathway for non-destructive
predominates. energy dissipation. Furthermore, the pyrimidine bases
are known to undergo free radical reactions more
readily than the purine bases. The order adenine >
Vitamins and coenzymes guanine > cytosine > uracil > thymine has been
established for the relative stabilities of the bases to
The irradiation of coenzyme I (diphosphopyridine radiolysis. In the presence of oxygen in aqueous sol-
nucleotide) results in reduction of the pyridine-carbox- ution, uracil and thymine form stable hydroperoxides,
amido ring. The product of this reduction is probably a whereas cytosine forms an unstable hydroperoxide that
dimer that is itself radiosensitive. decomposes to a variety of products.
The B group vitamins, thiamine and riboflavin, are Irradiation of DNA in the solid state at liquid
destroyed on irradiation in dilute aqueous solutions. nitrogen temperature yields radicals in which, electron
Riboflavin is reduced in air-free solutions to a semiqui- spin resonance measurements indicate, the unpaired
none form. Nicotinic acid is decarboxylated on spin is delocalized over the entire chain and does
irradiation in air-saturated aqueous solutions. not belong to any one unit of the giant molecule.
Sterilization: Radiation 3549

Sterilization
Statistical–
The addition of small amounts of water to this system Therefore, the most likely targets to be damaged
does not alter the nature of the DNA radicals pro- are those with the first two criteria. The two cellular
duced, but a two-to-one excess of water results in the components that best satisfy all three criteria are
annihilation of the electron spin resonance signal the genomic material (DNA) and the cytoplasmic
for DNA with the appearance of a strong signal membrane.
due to water radicals. It has been postulated that this
protective effect is attributable to energy transfer that
is made possible in an excess of water by structuring
of the water, thus providing a pathway for the forma- Evidence Supporting DNA
tion of a delocalized water radical, or exciton. Electron as a Critical Target
spin resonance studies of irradiated nucleoprotein
solutions indicate that the protein takes most of the DNA is considered as one of the plausible critical
radiation damage, protecting the nucleic acid moiety. targets for radiation action because there is only one
The damages caused by ionizing radiations in or a few copies of the genome in each cell, because it
nucleic acid and their components are obviously detri- is large compared with other molecular components,
mental to the passage of genetic information that and because it plays a critical role in the proliferation
requires specific order of intact purine and pyrimidine and survival of the cell.
bases in the DNA strands. Alterations in these bases The most compelling pieces of experimental evi-
and the DNA molecules in general can lead to muta- dence that DNA is a critical target for the biological
tions and lethal genes. The disruption of RNA mole- action of ionizing radiation are the mutagenic effects
cules interferes with protein synthesis and can result of ionizing radiation and the existence of DNA
in eventual cell death. repair-deficient mutants of bacteria and cultured mam-
malian cells, which display a high sensitivity to killing
by ionizing radiation.
Cellular genetic information is determined by the
RESPONSE OF BIOLOGICAL SYSTEMS base sequence of the genomic DNA. Any alterations
TO IONIZING RADIATION in this base sequence, such as damage to the bases in
DNA (see above), will result in changes to this base
Identification of the Critical Target sequence and, thus, to potentially mutagenic events.
Whether the base sequence change will result in a
As noted previously, absorption of ionizing radiation phenotypic change depends on exactly where the base
energy depends on the atomic weights of the atoms change occurs. If it is in an essential region of the
of the material and the density of the material. In con- DNA for the gene coding for a protein product or
trast to the absorption of ultraviolet and visible radi- some functional or structural RNA product (tRNA
ation, the absorption of ionizing radiation is virtually or rRNA), then the base change will give rise to a
independent of the nature of the molecular structure. phenotypic mutant. Otherwise, the mutation will fall
Thus, the release of ionizing radiation energy in into the category of a ‘‘silent mutation,’’ which refers
biological systems is essentially independent of the to those genotypic changes that do not have any
molecular bonds contained in the different biological associated phenotypic changes. For either type of
molecules. This complicates the task of identifying mutation, the DNA is the component that must
the critical actions of this type of radiation. A good undergo change for the mutagenic event to occur.
understanding of the action of ionizing radiation on An important advance in the understanding of
living cells requires that the biologically important how radiation kills biological systems involved the
events should be known. discovery that certain cells were more sensitive to
Determination of the critical sites for killing and killing by radiation than others. This discovery
mutagenesis by ionizing radiation has required biologi- was first observed with the killing of bacteria by ultra-
cal experiments designed to answer this question. violet radiation. In the case of ultraviolet radiation, it
Several criteria essential for a target to qualify as a was rather straightforward to establish that the bio-
critical site for radiation action include: 1) relatively chemical damage involved in this differential sensi-
large size; 2) one or only a few copies in the cell; and tivity of cells was in DNA and, more specifically,
3) that it should serve a critical function for the growth that it was the result of the formation of intrastrand
and survival of the cell. The first two criteria result (or, much less frequently, interstrand) cyclobutane
from the statistical distribution of damaging events in pyrimidine dimers. Several factors facilitated
the cell. From target theory, it has been shown that the identification of this type of damage. First, pyrimi-
the physical distribution of energy released from ioniz- dines absorb ultraviolet light strongly at 260 nm, and
ing radiation follows a Poisson statistical distribution. the efficiency of killing by ultraviolet radiation is
 3550 Sterilization: Radiation
Sterilization
Statistical–

maximal at or near this wavelength. Second, there was Relative Sensitivity of Various
available a sensitive and quantitative assay for this type Biological Systems to Killing
of damage. Thus, it was possible to quantitatively mea- by Ionizing Radiation
sure this biochemical damage and establish its relation
to biological killing by the ultraviolet radiation. To design a protocol for sterilization of a solution or
Several decades later (in the 1960s), the same differ- material by ionizing radiation, it is necessary to estab-
ential sensitivity of bacteria to killing by ionizing radi- lish the exposure dose required to kill all living organ-
ation was observed, and the biochemical damage isms present, which is the definition of the sterilization
responsible for this differential sensitivity was determ- dose. There is a large amount of data that provides
ined to be single-strand breaks in the backbone of the measures of the radiosensitivity of diverse biological
DNA. DNA strand breaks can be measured by a sensi- systems to radiation. For pharmaceuticals, it is impor-
tive assay, using agarose gel electrophoresis, thus tant to consider viruses as well as pathogenic bacteria
affording the opportunity to quantitatively relate the and other pathogenic organisms when determining
production of strand breaks with cell killing. Addition- the exposure dose required for sterilization of a parti-
ally, the isolation of radiation-sensitive mutants that cular product. It is also necessary to consider the
lacked the ability to efficiently repair strand breaks physical and chemical environmental factors that cause
was achieved. Thus, there was good evidence that this variations in radiation sensitivity when establishing
type of damage was related to the killing action of sterilization doses for the material, for the reasons that
ionizing radiation. have been addressed previously. In general, the most
radioresistant biological systems are the viruses. Cer-
tain bacterial species (e.g., Micrococcus radiodurans)
Evidence that the Cytoplasmic Membrane are as resistant as many viruses. The 37% survival
Is a Critical Target dose forthese radioresistant bacteria is approximately
20kGy (2 Mrad).
The cytoplasmic membrane is critical for the survival
of the cell. It is large, and there is only one present
Survival Curves for Microorganisms
for each cell. Therefore, the cytoplasmic membrane
has the key characteristics that make it a likely critical
Ultimately, the goal of ionizing radiation treatment of
target for killing by ionizing radiation. Experimental
pharmaceuticals is to kill all living organisms, that is,
evidence that the membrane plays a role in the lethal
to sterilize the product. To achieve this, it is important
action of ionizing radiation involves the observation
to understand and know the survival curve response of
that oxygen increases the radiosensitivity of cells to
microorganisms to killing by radiation. Fig. 2 displays
killing by ionizing radiation. This radiosensitization
the various kinds of survival curves that may be
by oxygen during radiation exposure is commonly
observed for microorganisms. No numbers have been
called the oxygen effect and is active in the killing of
assigned to the axes, because only the overall kinetic
bacteria, mammalian cells, and plant cells, but not
response of killing by the radiation is of interest.
viruses. Also, the biological inactivation of free DNA
Curves a and b show the killing action for two hypo-
(either viral or transforming DNA) by ionizing radi-
thetical organisms with the same rate of killing but
ation does not display the oxygen effect.
with differing repair capacities. Organisms capable of
As a result of the different responses to the oxygen
repairing lethal damage will have a characteristic
effect, it is common to refer to the killing action of
threshold dose below which there is complete survival
ionizing radiation including a component that is
oxygen-dependent and one that is oxygen-independent
damage. Each of these results in cell-killing. The
oxygen-independent damage is considered to be dam-
age that involves the DNA, and the oxygen-dependent
damage is considered to be that to the cytoplasmic
Log survival

membrane. Typically, the efficiency of killing by ioniz-

ing radiation is two to four times greater when
exposure occurs in the presence of oxygen compared
with when exposure occurs in an anoxic environment.
Thus, it is reasonable to predict that between 50 and
c b a
75% of the lethal action of ionizing radiation results d
from membrane damage. For the oxygen effect to Radiation dose
occur, the oxygen must be present during radiation
exposure. Fig. 2 Biological survival curves.
Sterilization: Radiation 3551

Sterilization
Statistical–
from radiation exposure. As the radiation dose increases, of sterilized pharmaceuticals, the two physical methods
the repair system is not able to completely repair all radi- under consideration are heat and ionizing radiation.
ation damage, and there is an exponential decrease in the Concerns similar to those associated with the use of
viability of the population with increasing radiation ionizing radiation in food processing have been
doses. Alternatively, requirement for multiple ionization raised. After extensive discussions and hearings, the
events in a critical target, or ionization events in multiple FDA has approved the commercial use of ionizing
targets, can give rise to a threshold dose below which radiation for processing fruits, vegetables, poultry,
there is no observed radiation killing. beef, and spices. Before this technique is approved by
Curves c and d show the killing of organisms that the FDA as a physical agent for sterilization of phar-
lack the ‘‘shoulder’’ portion of curves a and b. The maceuticals, similar hearings will be required. The pri-
slope of the survival curve for organism d is greater mary issue to be considered when developing a
than for organism c because organism d is more sensi- protocol using ionizing radiation is the establishment
tive to radiation killing than organism c. Two organ- of the minimum and maximum allowable exposure
isms may differ in both the values of their threshold doses. A minimum dose limit is required, to ensure that
doses and in their kinetics of inactivation by radiation. the product has been exposed to a sufficiently high
It is important to keep in mind the shape and rate of dose that results in sterilization of the product. A
inactivation of unwanted organisms when designing maximum dose limit is required to avoid damage to
protocols for radiation sterilization. the product resulting from unnecessarily high exposure
doses. Exposure dose limits in the FDA regulations for
food processing include minimum and maximum
USE OF IONIZING RADIATION IN acceptable dose limits, which were established with
THE PREPARATION OF STERILE these goals in mind.
PHARMACEUTICALS FOR Experiments on the action of ionizing radiation on
HUMAN USE pharmaceuticals and the killing of bacteria by ionizing
radiation indicate that this treatment has the potential
At this writing, there are no approved procedures for to be used in terminal sterilization. Reports indicate
the preparation of sterilized pharmaceuticals by that there are no products formed in irradiated
exposure to ionizing radiation, although several pack- samples of penicillin G, neomycin, novobiocin, and
aging materials are approved for sterilization by doses dihydrostreptomycin, which are different from those
in the 5- to 25-kGy dose region. The U.S. Food and that are formed by acidic, basic, hydrolytic, and oxi-
Drug Administration (FDA) is considering rules that dative treatments of these antibiotics. At the same
would permit the use of ionizing radiation in the ter- exposure doses, there was a 1 million-fold reduction
minal sterilization of drugs for human use. The two in the number of viable bacterial spores, which are
processes under consideration are aseptic processing the most radiosensitive forms of endospore forming
and terminal sterilization for the preparation of sterile bacteria. Thus, the use of ionizing radiation may pro-
pharmaceuticals. These two methods differ in the vide an alternative to heat or chemical treatment for
manner by which sterilization of the product is the sterilization of pharmaceuticals. This could provide
achieved. In the case of aseptic processing, the product a solution to the problem of sterilization of heat-
and the container are sterilized separately, and then sensitive drugs. Also, avoidance of chemical sterili-
final packaging of the product is carried out under zation removes the possibility of contamination by
aseptic conditions. When terminal sterilization is used, residues of the chemical that was used for sterilization.
the finally packaged product is sterilized. The It has been reported that irradiation of the antibiotic
FDA favors the terminal sterilization approach cefotaxime formed radiation products from impurities
because fewer failures occur when using this technique present in the cefotaxime sample. One must be aware
compared with the failure rate of the aseptic processing of the role of solvents and chemicals other than the
techniques. When using terminal sterilization, the step drug itself when considering radiolytic changes of a
of transfer of the product into the sterilized container drug during sterilization.
under aseptic conditions is avoided, thus removing
the possibility for product contamination during the
transfer step. It is recognized that it will not be practical CONCLUSIONS
to use terminal sterilization for certain pharmaceuti-
cals, since they might be sensitive to alteration and The purpose of this article is to provide the pharma-
inactivation by the sterilizing agent, whereas they can ceutical industry with an overview of the physical,
be sterilized by filtration, when in solution. chemical, and biological actions of ionizing radiation
During ongoing deliberations of which techniques on molecules of interest to the industry, as well as to
should be approved by the FDA for the preparation provide a current perspective on the prospects of the
 3552 Sterilization: Radiation
Sterilization
Statistical–

practical use of ionizing radiation as a physical agent BIBLIOGRAPHY

in the sterilization of drugs for human use. Advantages
of using employing ionizing radiation for pharamaceu- Alper, T. Cellular Radiobiology 1979.
Barbarin, N.; Rollmann, B.; Tilquin, B. Role of residual solvents
tical sterilization include: in the formation of volatile compounds after radiosteriliza-
tion of cefotaxime. Int. J. Pharm. 1999, 178, 203–212.
Chemical protection from radiation effects. Nucleonics 1960, 18,
 effectiveness on all microorganisms 76–81.
Derr, D., Radiation Processing. In What Is It? Where Is It Going?
 no rise in temperature during treatment ASTM Standardization News 1993, 25–27.
 consistent and reproducible protocols Ebert, M., Howards, A., Eds.; Radiation Effects in Physics,
 readily controlled and validated methods. Chemistry and Biology; Year Book Medical Publishers: Chi-
cago, 1963.
Pullman, B.; Pullman, A. Quantum Biochemistry; John Wiley &
Sons: New York, 1963; 267–283.
Although there is currently no approval by the FDA Radiation chemistry. Nucleonics 1961, 19, 37–68.
for ionizing radiation sterilization, there are active Schwarz, H.A. Radiation chemistry. Ann. Rev. Phys. Chem.
1965, 16, 347–374.
hearings being held by the agency on this matter. Stock, D.A.; Achey, P.M. Repair of single-strand
Considering the desire by the FDA to encourage the breaks in DNA from cultured leptidopteran cells
terminal sterilization method for sterilization of phar- exposed to gamma radiation. In Invertebrate Cell System
Applications; Mitsuhashi, J., Ed.; CRC Press, 1989; 1,
maceuticals and the fact the ionizing radiation has 45–61.
distinct advantages over other treatment methods for State of the art symposium: radiation chemistry. J. Chem. Ed.
use in terminal sterilization as presented above, it is 1981, 58, 82.
Swallow, A.J. Radiation Chemistry of Organic Compounds; Per-
expected that there will be approval of the use of this gamon Press: London, 1960.
physical agent in sterilization of drugs for human use Symposium on the development of radiation chemistry. J. Chem.
after regulatory hearings have provided sufficient Ed. 1995, 36.
Tiliquin, B.; Crucq, A.S. Les mecanismes chimiques de la radio-
information to allow identification of the appropriate sterilisation de medicaments solides. J. Chim. Phys. 1999, 96,
conditions under which this agent can be used. Recent 167–173.
developments in the application of ionizing radiation Use of aseptic processing and terminal sterilization, the prep-
aration of sterile pharmaceuticals for human and veterinary
for sterilization are reported on Web sites http:// use. Federal Register, 1991; 56, No. 198, 51354–51358, Octo-
www.iaea.org and http://www.fda.gov. ber, 11, 1991.
Super Disintegrants: Characterization and Function
Larry L. Augsburger
Department of Pharmaceutics, University of Maryland at Baltimore,
Baltimore, Maryland, U.S.A.

Suspensions
Albert W. Brzeczko

Super–
APC/Niro Inc., Columbia, Maryland, U.S.A.

Umang Shah
Pfizer Global Research and Development, Morris Plains,
New Jersey, U.S.A.

Huijeong Ashley Hahm

Office of Generic Drugs, U.S. Food and Drug Administration, Rockville, Maryland, U.S.A.

INTRODUCTION manipulation. Carboxymethylation is performed by react-

ing starch with sodium chloroacetate in an alkaline
Disintegrating agents are substances routinely included medium and neutralizing with a citric or acetic acid,
in tablet formulations and in some hard shell capsule a process known as a Williamson ether synthesis. This
formulations to promote moisture penetration and dis- synthesis yields carboxymethylation of about 25% of the
persion of the matrix of the dosage form in dissolution glucose units. The by-products, which include sodium
fluids. An oral solid dosage form should ideally chloride, sodium glycolate, and sodium citrate or acetate,
disperse into the primary particles from which it was are partially washed out. The particle sizes of the disinte-
prepared. Although various compounds have been grants are increased by the substitution and cross-linking
proposed and evaluated as disintegrants, relatively processes.[3]
few are in common usage today. Traditionally, starch Sodium starch glycolates are generally spherical, a
has been the disintegrant of choice in tablet formula- characteristic that accounts for their good flowability.[4]
tions, and it is still widely used. However, starch is Fig. 1 shows the scanning electron photomicrographs
far from ideal. For instance, starch generally has to of some of the commercially available sodium starch
be present at levels greater than 5% to adversely affect glycolates.
compactibility, especially in direct compression. More-
over, intragranular starch in wet granulations is not as
effective as dry starch. In more recent years, several Croscarmellose Sodium
newer disintegrants have been developed. Often called
‘‘super disintegrants,’’ these newer substances can be Croscarmellose sodium is derived from internally cross-
used at lower levels than starch. Because they can be linking a cellulose ether, sodium carboxymethylcellulose,
a smaller part of the overall formulation than starch, which is a water soluble polymer. It is composed of
any possible adverse effect on fluidity or compactibility repeating units of cellobiose units, with each unit
would be minimized. These newer disintegrants may consisting of two anhydroglucose units linked by 1,4-
be organized into three classes based on their chemical b-glucoside. Each unit also has three hydroxyl groups.
structure (Table 1). The degree of substitution refers to the average number
of hydroxyl groups substituted by carboxymethyl groups.
Croscarmellose sodium is made from crude cellulose,
GENERAL CHEMISTRY AND SURFACE which is steeped in sodium hydroxide solution.[1] The
MORPHOLOGY cellulose is subsequently reacted with sodium mono-
chloroacetate to form carboxymethylcellulose sodium.
Sodium Starch Glycolate After completion of the substitution, the excess sodium
monochloroacetate slowly hydrolyzes to glycolic acid.
Sodium starch glycolate is a super disintegrant made from The glycolic acid converts a few of the sodium carboxy-
cross-linking sodium carboxymethylstarch.[1,2] Cross- methyl groups to the free acid, catalyzes the cross-linkage
linking involves a chemical reaction with phosphorus to form croscarmellose sodium, and forms sodium
oxytrichloride or sodium trimetaphosphate, or a physical chloride and sodium glycolate as the by-products.
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-100001724
Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved. 3553
 3554 Super Disintegrants: Characterization and Function

Table 1 Classification of ‘‘super disintegrants’’ (partial listing)

Structural type (NF name) Description Trade name (manufacturer)
Modified starches Sodium carboxymethyl starch; the ExplotabÕ(Edward Mendell Co.)
(Sodium starch glycolate, NF) carboxymethyl groups induces PrimojelÕ (Generichem Corp.)
hydrophilicity and cross-linking TabloÕ (Blanver, Brazil)
reduces solubility.
AcDiSolÕ (FMC Corp.)
Suspensions

Modified cellulose Sodium carboxymethyl cellulose

(Croscarmellose, NF) which has been cross-linked to Nymcel ZSXÕ (Nyma, Netherlands)
Super–

render the material insoluble. PrimelloseÕ (Avebe, Netherlands)

SolutabÕ (Blanver, Brazil)
Cross-linked poly-vinylpyrrolidone Cross-linked polyvinylpyrrolidone; Crospovidone MÕ (BASF Corp.)
(Crospovidone, NF) the high molecular weight and Kollidon CLÕ (BASF Corp.)
cross-linking render the material Polyplasdone XL (ISP Corp.)
insoluble in water.

Most of the by-products can be removed to achieve the croscarmellose sodium particles are fibers with
99.5% purity by extraction with alcohol. Croscarmellose fairly sharp ends, probably because of the milling
sodium may be milled to break the polymer fibers into process (Fig. 2).
shorter lengths and hence improve its flow properties.
Unlike sodium starch glycolate, crude croscarmellose
sodium particles do not flow very well because of their Crospovidone
twisted fibrous morphology and varying lengths. There-
fore, they are cryogenically milled to improve flowability. Crospovidone is a cross-linked homopolymer of
The scanning electron photomicrographs show that N-vinyl-2-pyrrolidone. The reactants, acetylene and

Fig. 1 Scanning electron photomicrograph of sodium starch glycolates: (A) ExplotabÕ; (B) PrimojelÕ; and (C) TabloÕ
(600
magnification).
Super Disintegrants: Characterization and Function 3555

Suspensions
Super–
Fig. 2 Scanning electron photomicrograph of croscarmelloses: (A) AcDiSolÕ; (B) Nymcel ZSXÕ; (C) PrimelloseÕ; and (D)
SolutabÕ (100
magnification).

formaldehyde, are used to form butynediol. The hydro- DISINTEGRANT ACTION

genation and subsequent cyclodehydrogenation of
butynediol form butyrolactone. The reaction of butyro- Although disintegrants are important components in
lactone with ammonia produces pyrrolidone that is solid dosage forms, their mechanism of action has not
then vinylated with acetylene under pressure. The linear been clearly elucidated. The mechanisms proposed in
polymerization of the vinylpyrrolidone yields polyvinyl- the past include water wicking, swelling, deformation
pyrrolidone, a soluble binder, while the popcorn (bran- recovery, repulsion, and heat of wetting. It seems likely
ched) polymerization yields crospovidone, an insoluble that no single mechanism can explain the complex
super disintegrant. The by-products of popcorn poly- behavior of the disintegrants. However, each of these
merization include vinylpyrrolidone and polyvinyl- proposed mechanism provides some understanding of
pyrrolidone. Crospovidone contains less than 1.5% of different aspects of disintegrant action.
the soluble material, which has been determined to be
polyvinylpyrrolidone by infrared spectroscopy.
Crospovidone particles have a very different appear- Water Wicking
ance from those of the other two classes of super disin-
tegrants. Crospovidone particles seem to consist of The ability of a disintegrant to draw water into the
aggregates of smaller particles that are fused together. porous network of a tablet is essential for effective dis-
This aggregation gives crospovidone a spongy, highly integration. For crospovidone, water wicking has been
porous appearance (Fig. 3). Scanning electron photomi- thought to be the main mechanism of disintegration.
crographs show that the reduction of particle size of Kornblum and Stoopak[5] observed that crospovidone
crospovidone increases the surface area per unit weight, swells very little, yet takes water into its network quite
but decreases the intraparticulate porosity and the rapidly. Even the extensively swelling sodium starch
spongy appearance.[4] glycolate showed improved disintegration when the
 3556 Super Disintegrants: Characterization and Function
Suspensions
Super–

Fig. 3 Scanning electron photomicrograph of crospovidones: (A) Crospovidone MÕ; (B) Kollidon CLÕ; (C) Polyplasdone XLÕ;
and (D) Polyplasdone XL-10Õ (150
magnification).

molecular structure was altered to improve water Super disintegrants draw water into the matrix
uptake, as observed by Rudnic, Kanig, and Rhodes[6] system at a faster rate and to a greater extent when
Unlike swelling, which is mainly a measure of volume compared to traditional starch.[9] Van Kamp et al.[10],
expansion with accompanying force generation, water utilizing a water uptake measurement device, were able
wicking is not necessarily accompanied by a volume to show that tablets that demonstrate greater uptake
increase. volume and rate, such as those containing sodium
The ability of a system to draw water can be sum- starch glycolate, disintegrated more rapidly. Although
marized by Washburn’s equation:[7] the hydrophobic lubricant, magnesium stearate, seemed
  to negatively affect the wicking process, those containing
2 g cos y sodium starch glycolate were less affected by the detri-
L ¼ rt ð1Þ
2Z mental effect of mixing with the hydrophobic lubricant.
Lerk et al.[11] also observed a decreased rate of wetting
The Washburn equation is too simplistic to apply to a when disintegrants were mixed with magnesium stearate
dynamic tablet-disintegration process, but it does show for various mixing times. The decrease in the rate of
that any change in the surface tension (g), pore size (r), wetting was proportional to the time of mixing. Most
solid–liquid contact angle (y), or liquid viscosity (Z) likely, this observation reflected a greater delamination
could change the water wicking efficiency (L ¼ length of magnesium stearate at longer mixing times.
length of water penetration in the capillary; t ¼ time).
For example, when Rudnic et al.[8] evaluated the disinte- Swelling
gration efficiency of different particle sizes of crospo-
vidone, those with the largest particle size range Although water penetration is a necessary first step for
(50–300 mm) yielded the shortest disintegration time. disintegration, swelling is probably the most widely
Large particle size probably yielded greater pore size and accepted mechanism of action for tablet disintegrants.
altered the shape of the pore. Indeed, longer fiber length Indeed, most disintegrants do swell to some extent, but
due to greater particle size could improve the efficiency the variability of this property between disintegrants
of capillary uptake of water into the dosage form matrix. reduces its plausibility as a sole mechanism.
Super Disintegrants: Characterization and Function 3557

The earliest attempt to measure swelling was to pression, and that the particles return to their precom-
measure the sedimentation volume of slurries. Nogami pression shape upon wetting, thereby causing the tablet
et al.[12] developed a reliable test to measure both to break apart. Hess,[20] with the aid of photomicro-
swelling and water uptake. Gissinger and Stamm[13] graphs, showed that deformed starch particles returned
modified this apparatus and found a positive corre- to their original shape when exposed to moisture.
lation between the rate of swelling and disintegrant Fassihi[21] concluded that at higher compression
action for some disintegrants. List and Muazzam[14] forces, disintegration may become dependent on

Suspensions
later adapted this apparatus to measure both rate of mechanical activation of the tablet, resulting from the

Super–
swelling and swelling force through the application of stored energy imparted by the compression process.
force and displacement transducers. They found that He examined the disintegration times of tablets made
disintegrants that generate large swelling forces are of EmdexÕ powder, magnesium stearate, and 5% disin-
generally more effective. tegrant. Regardless of the disintegrant used (sodium
For swelling to be effective as a mechanism of disin- starch glycolate, microcrystalline cellulose, corscar-
tegration, there must be a superstructure against which mellose sodium, or starch), the disintegration time
the disintegrant swells. Swelling of the disintegrant increased with increasing compression force, then
against the matrix leads to the development of a swelling decreased again when the compression force was above
force. A large internal porosity in the dosage form in 120 MN/m2.
which much of the swelling can be accommodated Research on deformation and its recovery in situ
reduces the effectiveness of the disintegrant. At the as a disintegration mechanism is incomplete. However,
same time, a matrix that yields readily through plastic such a mechanism may be an important aspect of
deformation may partly accommodate any disintegrant the mechanism of action of disintegrants such as
swelling if swelling does not occur at a sufficient crospovidone and starch that appear to exhibit little
rapidity. or no swelling. The efficacy of such disintegrants likely
The swelling of some disintegrants is dependent on would be dependent on the relative yield strength of
the pH of the media. Shangraw, Mitrevej, and Shah[3] the disintegrant and that of the matrix in which it is
reported that sedimentation volumes of anionic cross- compressed, since disintegration efficiency would surely
linked starches and celluloses are altered in acidic depend on how much deformation is sustained by the
media. Polyplasdone XLÕ and Starch 1500Õ were disintegrant particles. Also, time-dependent stress relax-
unchanged. In a separate study, Chen et al.[15] showed ation could possibly be a factor in the aging of such
that acetaminophen tablets containing PrimojelÕ and tablets in that any deformation induced into the dis-
AcDiSolÕ had longer disintegration and dissolution integrant, that cannot be sustained by intraparticulate
times in acidic medium compared to neutral medium. bonding, gradually may recover as the matrix relaxes.
Those containing Polyplasdone XLÕ showed no such
differences. Mitrevej and Hollenbeck[16] verified the
Repulsion Theory
remarkable swelling capacity of some ‘‘super disinte-
grants’’ by exposing individual particles deposited on
Ringard and Guyot-Hermann[22] have proposed a
slides to high humidities and observing their degree of
particle–particle repulsion theory to explain the obser-
swelling microscopically.
vation that particles that do not swell extensively, such
On the other hand, when Caramella et al.[17–19] evalu-
as starch, could still disintegrate tablets. In this theory,
ated different disintegrants for their ability to swell, no
water penetrates into the tablet through hydrophilic
correlation could be observed between the maximum
pores and a continuous starch network that can convey
disintegrating force and percent of particle swelling.
water from one particle to the next, imparting a signifi-
Because they did observe a correlation between the rate
cant hydrostatic pressure. The water then penetrates
of disintegrating force development and the disintegra-
between starch grains because of its affinity for starch
tion time, therefore, the authors suggested that the rate
surfaces, thereby breaking hydrogen bonds and other
of development of a disintegrating force is all-important.
forces holding the tablet together. Presently, this theory
Swelling capable of rapid force development may be
is not supported by adequate data.
preferred since a slowly developing force could hypo-
thetically allow tablets to relieve the stress generated
without bond disruption. Heat of Wetting

Matsumara[23] noticed that starch particles exhibit

Deformation Recovery slight exothermic properties when wetted, which was
thought to cause localized stress resulting from expan-
The deformation recovery theory implies that the shapes sion of air retained in the tablet matrix. Unfortunately,
of the disintegrant particles are distorted during com- this explanation, if valid, would be limited to only a
 3558 Super Disintegrants: Characterization and Function

few substances such as aluminum silicate and kaolinite. axial and radial swelling forces. As indicated in
List and Muazzam[24] found that exothermic wetting Figs. 4–6, tablet compaction contributes more to the
reactions were not exhibited with all disintegrants axial pressure than to the radial pressure when super
and that even when a significant heat of wetting was disintegrants representing the three main super disinte-
generated, disintegration time did not always decrease. grant classes were studied in model tablet formula-
Caramella et al.[25] found that an increase in tempera- tions. In all three cases, the maximum axial pressure
ture, which should cause air expansion, did not enhance in an anhydrous lactose matrix was well below that
Suspensions

maximum force generation in several formulations. observed with a dicalcium phosphate dihydrate matrix
Super–

Therefore, they concluded that expansion of air in when the disintegrants are compared at the same con-
pores from heat of wetting could not be supported by centration. The differences in disintegrant performance
the data. More recently, Luangtana-anan et al. have in soluble and insoluble matrices could be rationalized
examined the heat of wetting of powders and tablets in terms of pressure development and liquid uptake.
of magnesium carbonate and EmcompressÕ.[26] Fig. 7 compares the maximum axial disintegrating
Magnesium carbonate tablets with significantly pressure and disintegration times of the tablets con-
higher heat of wetting disintegrated more readily than taining 2% of the disintegrants in a matrix composed
the EncompressÕ tablets. Indeed, it would be interest- of dicalcium phosphate or lactose.[27] As can be seen,
ing to develop a model for the mechanism of tablet a higher disintegration pressure favors rapid disinte-
disintegration using a thermodynamic approach; how- gration of the dicalcium phosphate-based tablets, but
ever, heat of wetting alone probably is inadequate to a slower disintegration of lactose-based tablets. Higher
explain disintegration. initial axial disintegrating pressure rates also yield
shorter disintegration times for the dicalcium phos-
phate-based tablets, but no such correlation is seen
with the lactose-based tablets, whose disintegrating
Generation of a Disintegrating Force pressure rates are much lower than those of the dical-
or Pressure as a Unifying Principle cium phosphate-based tablets (Fig. 8). Maximum
water uptake and water uptake rate seem to be poor
The rate of generation of a disintegrating force may predictors of disintegration time, as seen in Figs. 9
be a unifying factor in the mechanisms of disinte- and 10. However, lactose-based tablets show a trend
gration.[19] Many proposed mechanisms may be visua- toward slower disintegration with faster liquid uptake.
lized as giving rise to a force. Brzeczko[27] developed It was suggested that faster liquid uptake leads to a
techniques to simultaneously measure the rate of liquid faster dissolution of lactose and increased porosity to
uptake into a tablet and the rate of generation of both accommodate swelling and/or structural recovery.

A B

0.5
2
Disintegration Pressure (MPa)
Disintegration Pressure (MPa)

0.4
1.5
0.3

1
0.2

0.5 0.1

0 0

50 50
100 100
200 200

Compression Pressure (MPa) Compression Pressure (MPa)

Radial Pressure Axial Pressure Radial Pressure Axial Pressure

DiTab Anhydrous Lactose

Fig. 4 The effect of compression pressure on the axial and radial disintegrating pressures of compacts made with lactose or
DitabÕ, and AcDiSolÕ (5%). (From Ref.[27].)
Super Disintegrants: Characterization and Function 3559

A B

1.2

Disintegration Pressure (MPa)

Disintegration Pressure (MPa)

0.35
1 0.3

0.8 0.25

Suspensions
Super–
0.6 0.2

0.15
0.4
0.1
0.2
0.05
0 0
50 50
100 100
200 200
Compression Pressure (MPa) Compression Pressure (MPa)
Radial Pressure Axial Pressure Axial Pressure Radial Pressure

DiTab Anhydrous Lactose

Fig. 5 The effect of compression pressure on the axial and radial disintegrating pressures of compacts made with lactose or
Ditab, and PrimojelÕ (5%). (From Ref.[27].)

Peppas[28] attribute the difference in disintegration Here, F is the disintegration force at time t, Fo is the
rate between soluble and insoluble matrices to two maximum force developed, k is an expansion rate
proposed phenomena—an interface-controlled mech- constant, and n signifies which of the two mechanisms
anism and a diffusion-controlled mechanism—as controls the disintegration. The interface controlled
represented in the following equation: phenomenon involves tablet particles breaking apart
from the interface of the tablet and the diffusion-
F=F1 ¼ 1  expðktn Þ ð2Þ controlled phenomenon involves particles diffusing

A B
Disintegration Pressure (MPa)

1.2
0.4
Disintegration Pressure (MPa)

1
0.3
0.8

0.6 0.2

0.4
0.1
0.2

0 0
50 50
100 100
200 200
Compression Pressure (MPa) Compression Pressure (MPa)
Radial Pressure Axial Pressure
Radial Pressure Axial Pressure
DiTab Anhydrous Lactose

Fig. 6 The effect of compression pressure on the axial and radial disintegrating pressures of compacts made with lactose or
DitabÕ, and Polyplasdone XLÕ (5%). (From Ref.[27].)
 3560 Super Disintegrants: Characterization and Function

Maximum Water Uptake

Maximum Axial Disintegrating 120
Dicalcium Phosphate
1.0 100 Lactose
0.9 Dicalcium Phosphate
Pressure (MPa)

0.8 Lactose 80
0.7

(g)
0.6 60
0.5
40
0.4
0.3
Suspensions

20
0.2
Super–

0.1 0
0.0 0 10 20 30 40 50
0 10 20 30 40 50 Disintegration Time (sec)
Disintegration Time (sec)
Fig. 9 Maximum water uptake versus disintegration time of
Fig. 7 Maximum axial disintegrating pressure versus dicalcium phosphate and lactose tablets containing 2% super
disintegration time of dicalcium phosphate and lactose disintegrants. 
¼ Dicalcium phosphate: no significant
tablets containing 2% super disintegrants. ¼ Dicalcium  correlation. ` ¼ Lactose: no significant correlation. (From
phosphate: r 2 ¼ 0.92, p < 0.05, significant correlation. Ref.[27].)
` ¼ Lactose: r 2 ¼ 0.81, p < 0.05, significant correlation.
(From Ref.[27].)
for the system containing b-lactose. In other words, the
super disintegrant’s interface-controlled mechanism could
away. Although it is thought that both happen simul- not overcome the diffusion-controlled mechanism of the
taneously, the degree to which disintegration depends b-lactose.[29]
on each system can differ. For example, those tablet
matrices with a relatively small n of about 0.6 are
thought to be dominated by the diffusional mechanism;
whereas, those with an n of greater than 0.9 are thought FACTORS AFFECTING
to be interfacial mechanism dominant. The value of n DISINTEGRANT ACTIVITY
would certainly differ based on the solubility of the
matrices. Particle Size
Since super disintegrants are highly hydrophilic yet
insoluble in water, they would be expected to be more Both the rate and force of disintegrant action may be
effective in breaking the tablet apart interfacially than dependent upon the particle size of the disintegrant.
controlling the diffusion per se. Indeed, Caramella et al. Smallenbroek, Bollhuis, and Lerk[30] found that starch
observed that disintegration occurred readily for tablets grains having relatively large particle sizes were more
containing insoluble calcium phosphate; whereas, tablets efficient than the smaller particle size grades. This is
containing highly soluble b-lactose disintegrated slowly. probably because the continuous hydrophilic network
Such phenomena were explained by a lower value of n of disintegrants is more efficiently accomplished by
the bigger particles. Also, Rudnic et al.[8] found that
coarser grades of crospovidone (50–100 mm, Grade B;
Pressure Rate (MPa/sec x 10–3)
Initial Axial Disintegrating

90
80 Dicalcium Phosphate 3.5
70 Lactose
Initial Water Uptake

3.0
60
Rate (mg/sec)

50 2.5
40 2.0
30
1.5
20
10 1.0
Dicalcium Phosphate
0 0.5 Lactose
0 10 20 30 40 50
0.0
Disintegration Time (sec) 0 10 20 30 40 50
Disintegration Time (sec)
Fig. 8 Initial axial disintegrating pressure rate versus disin-
tegration time of dicalcium phosphate and lactose tablets Fig. 10 Initial water uptake rate versus disintegration time
containing 2% super disintegrants. ¼ Dicalcium phos-  of dicalcium phosphate and lactose tablets containing 2%
phate: r 2 ¼ 0.95, p < 0.05, significant correlation. super disintegrants. 
¼ Dicalcium phosphate: no signifi-
` ¼ Lactose: r 2 ¼ 0.10, p < 0.05, no significant corre- cant correlation. ` ¼ Lactose: r 2 ¼ 0.47, p < 0.05, no sig-
lation. (From Ref.[27].) nificant correlation. (From Ref.[27].)
Super Disintegrants: Characterization and Function 3561

50–300 mm, Grade C) were more efficient than the finer and low levels of carboxymethylation was best for
particles (0–15 mm, Grade A). The differences in disin- tablet disintegration.
tegration efficiency between Grade B and Grade C
were not clear, however. When List and Muazzam[24]
Effect of Compression Force
evaluated two different grades of crospovidone particles
(100–200 mm and >315 mm), the efficiencies between the
Compression force affects disintegration time in
two grades were very similar (Table 2). Results for

Suspensions
different ways. First, it governs the penetration of
the other disintegrants, Amberlite IRP88Õ and potato

Super–
dissolution fluids into the matrix by controlling the
starch, support that coarser particle sizes allow more
porosity of the compact. Low compression force can
efficient disintegration than finer particles. For disinte-
lead to relatively high tablet porosity and can allow
grants that swell extensively, such efficiency can be
rapid penetration of water. However, it has often been
explained by the observed force development. Indeed,
observed that tablets containing starch exhibit disinte-
larger particles of sodium starch glycolate swelled more
gration times that tend to pass through minimum as
rapidly and to a greater extent than did the smaller
compression force increases.[21] At low compression
particles.[6]
forces, any possible swelling or deformation recovery
that may take place may be more or less accommo-
Molecular Structure dated by the porosity, whereas at intermediate com-
pression forces, a maximal disintegrating effect may
Disintegrants can vary in molecular structure based on develop. At high compression forces, fluid penetration
how they are manufactured or processed. Corn starch, may be impeded by a further reduction of porosity while
for example, contains different ratios of two sugar particle deformation of the disintegrants becomes more
fractions, amylose and amylopectin. Schwartz and important. In general, List and Muazzam[14] found
Zelinske[31] concluded that the linear polymer, amylose, increased swelling pressures at higher compression forces
was responsible for the disintegrant properties associa- when various amberlite resins, starches, and crospovi-
ted with starch, whereas the branched polymer, amylo- dones were employed at the 2.5% level in dicalcium
pectin, was responsible for the gummy property. phosphate matrix tablets (Table 2). Similar findings
Varying the amylose to amylopectin ratio did not were reported by and Fassihi[21] and Brzeczko.[27]
affect the porosities of the resulting tablets. Rudnic, In two different studies Khan and Rhodes[33,34]
Kanig, and Rhodes[6] evaluated the effect of cross- observed that tablets containing sodium starch glyco-
linking and carboxymethyl substitution in sodium late disintegrate relatively slowly at low compression
starch glycolate and concluded that the swelling of force, quickly at intermediate compression force, and
the disintegrant was largely inversely proportional to slowly again at high compression force. However, the
the degree of cross-linkage. Swelling also was inversely effects of compression force on the disintegration time
proportional to the level of substitution, but to a lesser of other types of disintegrants, such as cation exchange
degree. Shah, Bekersky, and Jarowski[32] found that resin, calcium sodium alginate, and various forms
carboxymethyl cellulose having high molecular weight of starches, varied widely. Perhaps the effect of

Table 2 Effect of particle size and compression pressure on swelling pressure and disintegration time
Compression Particle Swelling Disintegration
Disintegrant pressure (bar) size (lm) pressure (bar) time (s)
Amberlite IRP88Õ 625 <50 0.660 84
1560 <50 1.121 30
Amberlite IRP88Õ 625 100–200 1.083 52
1560 100–200 2.262 22
Potato starch 625 <50 0.165 254
1560 <50 0.310 164
Potato starch 625 80–100 0.234 160
1560 80–100 0.445 77
Polyplasdone XLÕ 625 100–200 0.898 31
1560 100–200 1.772 14
Polyplasdone XLÕ 625 >315 0.760 42
1560 >315 1.480 17
Tablet composition: 2.5% disintegrant, 1% magnesium stearate, and EmcompressÕ. (From Ref.[24].)
 3562 Super Disintegrants: Characterization and Function

slight postcompression expansion, which could explain

a slightly increased disintegration time compared to
the 5% disintegrant level.
The effect of compression force on disintegration
2400 efficiency seems, therefore, largely dependent on the
mechanism of the disintegrant action. The effectiveness
2000 of swelling or structure recovery may well be depen-
Suspensions

dent on attaining a compression force that achieves a

Super–

1600
critical porosity in the matrix. On the other hand, the
capillary uptake of liquid, which is a necessary precur-
D.T.(sg)

1200
sor to these mechanisms could be compromised if the
800 tablet matrix is compressed to too low a porosity.
300
260
400 220
180

)
Pa
0 140 Matrix Solubility

M
0

P(
2 100
4 6
8 10
The disintegrant mechanism seems to depend not only
%D
on the disintegrant itself but on the matrix as well.
Fig. 11 Surface response of disintegration time as functions of Disintegrants work most effectively in insoluble
compression pressure and percent disintegrant. (From Ref.[11].) matrices.[27] Insoluble matrices, such as those contain-
ing calcium phosphate do not disintegrate adequately
without disintegrants. On the other hand, tablets and
compression force on the disintegration time depends on capsules that primarily consist of water soluble fillers
the nature of the disintegrants, such as their mechanism or drugs tend to dissolve rather than to disintegrate,
of disintegration and deformation characteristics. even when disintegrating agents are present. It has
Munoz et al.[35] found that the effect of compression been suggested that during the dissolving process, the
pressure on disintegration time varied depending on the water acts as a plasticizer,[37] which can potentially
concentration used of the super disintegrant, ExplotabÕ. reduce the development of disintegrating force. In
Fig. 11 shows that shortest disintegration time could be addition, soluble materials that tend to swell can form
achieved at around 7% disintegrant concentration. At viscous plugs, which may impede further penetration
this concentration, compression force has little effect of moisture into the matrix. However, the addition of
on disintegration time. The disintegration time was more disintegrants almost predictably shortens disintegration
affected by compression force at low disintegrant time, despite the solubility of the matrix.
concentration, showing fastest disintegration time at
intermediate compression force. This type of biphasic
effect of compression force on disintegration time also Method of Incorporation in Granulation
was observed for AcDiSolÕ,[36] and the surface response
curve is very similar to that of ExplotabÕ. When disinte- The method of incorporation of disintegrants in granu-
gration times were studied at 5 and 10% disintegrant, lation has been controversial. Should the disintegrant
5% AcDiSolÕ yielded the lowest porosity, lowest yield be all extragranular, all intragranular, or divided between
pressure in Heckel analysis, and shortest disintegration these two locations? Shotton and Leonard[38] reported
time. At a 10% disintegrant level, the tablets showed a that maize starch, sodium calcium alginate, alginic acid,

Table 3 Effect of method of disintegrant addition in granules on the tablet properties

Crushing strength (kgf) Disintegration time (s)
Disintegrant
addition method Intra Equal Extra Intra Equal Extra
Control 6.5 664
4% PrimojelÕ 5.3 5.0 5.8 38 41 49
4% Ac-Di-SolÕ 3.8 4.8 5.7 110 126 148
4% Nymcel zsd 16Õ 4.0 4.3 6.5 499 540 488
4% Polyplasdone XLÕ 5.8 6.0 6.1 31 40 43
20% Potato Starch 3.3 3.4 2.1 69 80 110
(From Ref.[39].)
Table 4 Comparison of different modes of incorporation of disintegrants in granules on disintegration efficiency of tablets, as reported by different investigators
Drug (D), Order of
Binder (B), Disintegrants disintegration Additional
Super Disintegrants: Characterization and Function

Investigators Filler evaluated efficiency comments

Shotton and Leonard Sulfadiazine (D) Maize starch Extra > Intra Extragranular incorporation yielded
Povidone (B) Na calcium alginate fastest disintegration, but intragranular
No filler Alginic acid incorporation yielded finer particles.
Microcrystalline cellulose Equal distribution of disintegrants
Colloidal aluminum silicate is recommended.
Van Kamp, Bolhuis, and Lerk Prednisone (D) Potato starch Intra > Extra The difference between the modes of
Gelatin (B) incorporation for super disintegrants
Lactose (F) is not great.
PrimojelÕ Intra > Equal > Extra
Polyplasdone XLÕ Intra > Equal > Extra
NymcelÕ Extra > Intra > Equal
Khattab, Menon, and Sakr Paracetamol (D) Croscarmellose Na Equal > Extra > Intra Overall, the equal distribution of
Polyvinylpyrrollidone (B) disintegrants yielded the fastest
No filler disintegration and dissolution.
Na starch glycolate Equal > Intra or Extra
Crospovidone Equal > Intra > Extra
(From Refs.[38,39,41].)
3563

Super–
Suspensions
 3564 Super Disintegrants: Characterization and Function

and other disintegrants gave more rapid disintegration substituted hydroxypropylcellulose (L-HPC) than
when incorporated extragranularly than when incorpor- when polyvinylpyrrolidone or hydroxypropylmethyl-
ated intragranularly in a sulfadiazine granulation. They cellulose was the binder. In addition, the difference
also reported that the latter method gave a finer disper- seen in the effectiveness of starch in different modes
sion and concluded that the best compromise was to use of incorporation between the Shotton and Leonard[38]
both intra- and extragranular disintegrant. study and the Van Kamp, Bolhuis and Lerk[39] study
Van Kamp, Bolhuis, and Lerk[39] evaluated the may be related to the absence or presence of lactose,
Suspensions

method of incorporation of PrimojelÕ, AcDiSolÕ, a soluble filler. Unlike the Shotton and Leonard study,
Super–

and Polyplasdone XLÕ in prednisone tablets formed Van Kamp, Bolhuis, and Lerk used lactose as a soluble
from lactose granules. Whether the incorporation of filler, which might have reduced the relative effective-
the super disintegrant was intragranular, extragranu- ness extragranular starch, making the intragranular
lar, or evenly distributed in both sites, they found incorporation more favorable.
little or no difference in disintegration time, crushing The observations summarized in Table 4 make any
strength, or dissolution of prednisone. Interestingly, attempt to generalize that one method of incorporation
their results with potato starch showed differences that of disintegrant in granulation is better than another
did not agree with the earlier work of Shotton and difficult. However, when all of the data are taken
Leonard[38] in that intragranular starch was more together, it would appear that the combined addition
effective than extragranular starch (Table 3). of disintegrants both extragranularly and intragranu-
Gordon, Chatterjee, and Chowhan[40] reported that larly would provide the best opportunity for optimal
dissolution of naproxen, a poorly soluble drug at disintegrant activity.
gastric pH levels, was faster when AcDiSolÕ was incor-
porated intragranularly, compared to when it was
incorporated extragranularly or evenly distributed Effect of Reworking
between the intra- and extragranular portions. Even
more recently, a study reported by Khattab, Menon, The effect of recompressing a wet massed microcrystal-
and Sakr[41] showed that the combined incorporation line cellulose matrix containing super disintegrants on
of intra- and extragranular disintegrating agents (sodium swelling force kinetics also has been considered.[43]
starch glycolate, croscarmellose sodium, or crospo- When the disintegrants were placed extragranularly,
vidone) in a paracetamol granulation resulted in faster only ExplotabÕ among those considered retained good
disintegration and dissolution than either extragranular efficiency after reworking. When placed intragranularly,
or intragranular incorporation alone. all disintegrants had reworking efficiencies equivalent to
More studies are necessary to elucidate the effect of that of the nondisintegrant control. Adding 2% disinte-
other factors, such as the type of binder, the type of grant extragranularly prior to the second compression
filler, and the solubility of the matrix, which may sig- restored disintegrant behavior for Polyplasdone XLÕ,
nificantly affect the effectiveness of disintegrants in but only partial restoration was seen for AcDiSolÕ.
different modes of incorporation. For example, Becker, In further work,[44] reworked tablets containing 2%
Rigassi, and Bauer-Brandl[42] found that extragranular disintegrant extragranularly were studied. The data in
crospovidone was more effective in an acetaminophen Table 5 illustrate that maximal swelling forces were
tablet when the binder was maltodextrin (Licab DSHÕ), reduced in all cases, but there was no correlation with
pregelatinized maize starch (Lycab PGSÕ), or low tablet disintegration time.

Table 5 Rework efficiency (%RE) of super disintegrants

Disintegrant Relative Fs Relative Fs
(2%) (35% porosity)a (40% porosity)a % REb
Control 0.842 0.848 45
Õ
Polyplasdone XL 0.941 0.926 64
ExplotabÕ 0.737 0.863 86
Õ
AcDiSol 1.045 0.951 45
a Maximum swelling force (1st compression)
Re  Fs ¼
Minimum swelling force (2nd compression)
b AUC (1st compression)
%RE ¼
AUC (2nd compression)
AUCs are the area under curve from the disintegration time versus compression pressure graphs.
(From Ref.[44].)
Super Disintegrants: Characterization and Function 3565

Incorporation in Hard Gelatin Capsules multifactorial study, all main factors, including disinte-
grant type, compression force, level of lubricant, and
The utility and performance of super disintegrants in filler type, were found to have significant effects on
direct-fill powder formulations for hard shell capsules dissolution (Figs. 12 and 13).[46] In most cases at lower
filled on tamping machines are roughly analogous to disintegrant concentration, increasing the tamping
those of direct compression tablet formulation. In a force improved the dissolution of hydrochlorothiazide,
study where capsules were filled under controlled most likely due to reduced porosity. When the filler was

Suspensions
tamping force conditions using an instrumented Zanasi changed from lactose to dicalcium phosphate, the

Super–
LZ 64 dosator machine, a dicalcium phosphate based magnitude and the order of effectiveness of the disinte-
formulation containing hydrocholorothiazide and dif- grants changed.
ferent super disintegrants were tested for dissolution Like the experience with tablets, the effect of disin-
times.[45] The croscarmelloses were found to be more tegrants in already rapidly soluble capsule matrices is
effective than sodium starch glycolate in promoting lower than that in water insoluble matrices. Perhaps
hydrochlorothiazide dissolution, whereas crospovi- doubling the concentration normally required for
done was the poorest in this regard. In a follow-up tablets is needed to effect efficient disintegration and
significantly affect dissolution. This need for higher

100
100

90
90
0.5% Mg Stearate
6% AcDiSol

80
6% Primojel

0.5% Mg Stearate
1% Mg Stearate

80

6% Primojel
4% AcDiSol

4% Primojel

31.7 kg

6% AcDiSol
22.6 kg
10.1 kg
% Labeled Content Dissolved in 30 Minutes

70 4% AcDiSol
% Labeled Content Dissolved in 30 Minutes

70

31.7 kg
22.6 kg
60
60

1% Mg Stearate
10.1 kg
Control

4% Primojel

50
50

40 40

30 30
Control

20 20
Compr Force

Compr Force
Disintegrant

Disintegrant
Lubricant

Lubricant

10 10

0 0

Fig. 12 The averaged effect of disintegrant, compression Fig. 13 The averaged effect of disintegrant, compression
force, and lubricant on the release of hydrochlorothiazide force, and diluent on the release of hydrochlorothiazide from
from anhydrous lactose based capsules. Control ¼ 0% disin- dicalcium phosphate based capsules. Control ¼ 0% disinte-
tegrant. (From Ref.[47].) grant. (From Ref.[47].)
 3566 Super Disintegrants: Characterization and Function

disintegrant concentration probably reflects the greater precursor to all other mechanisms. Not all mechanisms
pore structure of capsule plugs compared to com- are well supported by research. Disintegrants appear to
pressed tablets. At equivalent concentrations in model function by multiple mechanisms, but a predominant
lactose or dicalcium phosphate-based systems, sodium mechanism seems to be characteristic of each disinte-
starch glycolate and croscarmellose sodium were more grant type. Regardless of their validity, all proposed
effective than crospovidone in promoting dissolution mechanisms at least have the potential to generate a
of hydrochlorothiazide from capsules manufactured disintegrating force within the matrix and this appears
Suspensions

with the same tamping force.[47] For either filler, to be a unifying concept.
Super–

disintegration times and swelling correlated well with

dissolution.
ACKNOWLEDGMENT
NEW DISINTEGRANTS
The authors wish to express sincere appreciation and
gratitude to Ms. Lynn DiMemmo, FMC Co., Princeton,
Gellan gum and Xanthan SMÕ appear to have per-
N.J., for the SEM’s of the super disintegrants.
formance characteristics similar to those of super disin-
tegrants. Gellan gum is an anionic polysaccharide of
linear tetrasaccharides, and is derived from Pseudo-
monas elodea.[48] When 4% gellan gum was incor- REFERENCES
porated in an ibuprofen tablets, its disintegration
time was 4 min, which was much superior than that 1. Handbook of Pharmaceutical Excipients, 2nd Ed.;
American Pharmaceutical Association: Washington, DC,
obtained using dried starch or Avicel PH 102Õ 1994; 141–144, 462–466.
(>15 min), and comparable to those of ExplotabÕ, 2. Bolhuis, G.K.; Van Kamp, H.V.; Lerk, C.F. Effect of vari-
AcDiSolÕ, and Kollidon CLÕ (4–7 min). ation of degree of substitution, crosslinking, and purity of
the disintegration efficiency of sodium starch glycolate.
Xanthan SMÕ is a new USP xanthan derivative with Acta Pharm. Tech. 1984, 30, 24–32.
higher hydrophilicity and lower gelling tendency.[49] In 3. Shangraw, R.F.; Mitrevej, A.M.; Shah, M.N. A new era of
aspirin tablets with 3% Xanthan SMÕ disintegration tablet disintegrants. Pharm. Tech. 1980, 4, 49–57.
4. Shah, U.S. Evaluation of the Functional Equivalence of
time was about 10 min. Increasing the concentration Different Sources of Super Disintegrants in Pharmaceutical
of disintegrant beyond 3% did not improve the disinte- Tablets. Dissertation, University of Maryland: Baltimore,
gration time, whereas the most effective concentration 1996; 16–26, 92–112.
5. Kornblum, S.S.; Stoopak, S.B. A new tablet disintegrating
of AcDiSolÕ was 5%, yielding a disintegration time of agent: cross-linked polyvinylpyrrolidone. J. Pharm. Sci. 1973,
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ecular structure on the function of sodium starch glycolate
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303–320.
7. Washburn, E.W. The dynamic of capillary flow. Phys. Rev.
1921, 17, 273–283.
CONCLUSIONS 8. Rudnic, E.M.; Lausier, J.M.; Chilamkarti, R.N.; Rhodes,
C.T. Studies of the utility of cross-linked polyvinylpyrroli-
In summary, super disintegrants are excipients used done as a tablet disintegrant. Drug Dev. Ind. Pharm.
1980, 6, 291–309.
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forms to aid dissolution in vivo. Commonly used super compression of pharmaceuticals. Pharm. Ind. 1984, 46,
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10. Van Kamp, H.V.; Bolhuis, G.K.; De Boer, A.H.; Lerk,
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differ from traditional starch in that they are effective at disintegration. Pharm. Acta Helv. 1986, 61, 22–29.
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K. Interaction of tablet disintegrants and magnesium stea-
provides formulation scientists greater flexibility, parti- rate during mixing. II. Effect on dissolution rate. Pharm.
cularly in designing direct compression tablets. However, Acta Helv. 1982, 57, 282–286.
the effectiveness of both starch and super disintegrants 12. Nogami, H.; Nagai, T.; Fukuoka, E.; Sonobe, T. Disinte-
gration of the aspirin tablets containing potato starch and
depends heavily upon the composition of the tablet microcrystalline cellulose in various concentrations. Chem.
matrix, compression pressure, and in the case of granu- Pharm. Bull. 1969, 17, 1450–1455.
lation, the method of incorporation. 13. Gissinger, D.; Stamm, A. A comparative evaluation of the
properties of some tablet disintegrants. Drug Dev. Ind.
Because of the complexities involved, the mechanism Pharm. 1980, 6, 511–536.
of action of super disintegrants is not well understood. 14. List, P.H.; Muazzam, U.A. Swelling—the driving force
Some of the proposed mechanisms include water wick- behind disintegration. Pharm. Ind. 1979, 41, 1075–1077.
15. Chen, C.R.; Lin, Y.H.; Cho, S.L.; Yen, S.Y.; Wu, H.L.
ing, swelling, deformation recovery, particle repulsion, Investigation of the dissolution difference between acidic
and heat of wetting. Water uptake is a necessary and neutral media of acetaminophen tablets containing
Super Disintegrants: Characterization and Function 3567

super disintegrant and a soluble excipient. Chem. Pharm. 37. Murthy, K.S.; Ghebre-Sellassie, I. Current perspectives on
Bull. 1997, 45, 509–512. the dissolution stability of solid oral dosage forms. J.
16. Mitrevej, A.; Hollenbeck, R.G. Photomicrographic analysis Pharm. Sci. 1993, 82, 113–126.
of water vapor sorption and swelling of selected super 38. Shotton, E.; Leonard, G.S. Effect of intragranular and
disintegrants. Pharm. Tech. 1982, 6, 48–50. extragranular disintegrating agents on particle size of disin-
17. Caramella, C.; Columbo, P.; Conte, U.; Gazzaniga, A.; tegrated tablets. J. Pharm. Sci. 1976, 65, 1170–1174.
LaManna, A. The role of swelling in the disintegration pro- 39. Van Kamp, H.V.; Bolhuis, G.K.; Lerk, C.F. Improvement
cess. Int. J. Pharm. Tech. Prod. Mfr. 1984, 5, 1–5. by super disintegrants of the properties of tablets contain-
18. Caramella, C.; Columbo, P.; Conte, U.; LaManna, A. ing lactose, prepared by wet granulation. Pharm. Weekbld.

Suspensions
Swelling of disintegrant particles and disintegrating force Sci. Ed. 1983, 5, 165–171.
of tablets. Labo-Pharma. Probl. Tech. 1984, 32, 115–119. 40. Gordon, M.S.; Chatterjee, B.; Chowhan, Z.T. Effect of the

Super–
19. Caramella, C.; Columbo, P.; Conte, U.; LaManna, A. mode of croscarmellose sodium incorporation on tablet dis-
Tablet disintegration update: the dynamic approach. Drug solution and friability. J. Pharm. Sci. 1990, 79, 43–47.
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21. Fassihi, A.R. Mechanisms of disintegration and compact- 45, 687–691.
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J. Pharm. 1986, 32, 93–96. binders in wet granulation: a comparison using model for-
22. Guyot-Hermann, A.M.; Ringard, J. Disintegration mecha- mulations of different tabletability. Drug Dev. Ind. Pharm.
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particle–particle repulsive force. Drug Dev. Ind. Pharm. 43. Gould, P.L.; Tan, S.B. The effect of recompression on dis-
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23. Matsumara, H. Studies on the mechanism of tablet com- Drug Dev. Ind. Pharm. 1985, 11, 441–460.
pression and disintegration. IV. Evolution of wetting heat 44. Gould, P.L.; Tan, S.B. The effect of recompression on the
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24. List, P.H.; Muazzam, U.A. Swelling—a driving force in 45. Botzolakis, J.E.; Small, L.E.; Augsburger, L.L. Effect of
tablet disintegration. Pharm. Ind. 1979, 41, 1075–1077. disintegrants on drug dissolution from capsules filled on a
25. Caramella, C.; Ferrari, F.; Conte, U.; Gazzaniga, A.; dosator-type automatic capsule-filling machine. Int. J.
LaManna, A.; Colombo, P. Experimental evidence of disin- Pharm. 1982, 12, 341–349.
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26. Luangtana-anan, M.; Catellani, P.L.; Colombo, P.; hard-gelatin capsules. J. Pharm. Pharmacol. 1984, 36, 77–84.
Dinarvand, R.; Fell, J.T.; Santi, P. The role of bond weak- 47. Botzolakis, J.E.; Augsburger, L.L. Disintegrating agents in
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Pharm. Biopharm. 1992, 38, 169–171. Dev. Ind. Pharm. 1988, 14, 29–41.
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Tablet Disintegration Processes, University of Maryland: maceutical dosage forms. Drug Dev. Ind. Pharm. 1997, 23,
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Pharm. 1989, 51, 77–83. Investigation on a new modified USP xanthan with tablet-
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1975, 64, 166–168. action. Pharm. Tech. 1984, 8, 50–63.
34. Khan, K.A.; Rhodes, C.T. Effect of variation in compac- Mahmoud, H.M.; El-Shaboury, M.H. Effect of certain tablet
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Jimemez-Castellanos, M.R. Effect of explotabÕ on the disintegration. I. The effect of penetrating rate on tablet
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Pharm. 1998, 24, 785–791. Pesonen, T.; Paronen, P.; Ketolainen disintegrant properties of
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Jimemez-Castellanos, R. Disintegrating efficiency of cros- 139–147.
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 Supercritical Fluid Technology in Pharmaceutical Research
Aaron S. Mayo
Uday B. Kompella
Department of Pharmaceutical Sciences, University of Nebraska Medical Center,
Omaha, Nebraska, U.S.A.
Suspensions
Super–

INTRODUCTION HISTORY OF SCF TECHNOLOGY (FOOD

PROCESSING TO PHARMACEUTICALS)
A supercritical fluid (SCF) is a substance whose tem-
perature and pressure are simultaneously above its Industrial applications[1,2] of supercritical fluids had
critical point. Critical temperature (Tc) is the highest humble beginnings. The observation of the supercriti-
temperature at which a gas can be converted to a liquid cal phase was first cited in 1822 by Cagniard de la
by an increase in pressure. Critical pressure (Pc) is the Tour.[3] In the said work, the disappearance of the
highest pressure at which a liquid can be converted to a gas–liquid boundary by visual inspection of a sealed
traditional gas by an increase in the liquid temperature. glass container at elevated temperatures was noted.
The phase diagram of a pure substance illustrating the Following this, in 1879, Hannay and Hogarth[4]
supercritical region is depicted in Fig. 1. In practice, reported the ability of an SCF to dissolve low-pressure
substances at a reduced temperature (T/Tc) and solid materials. They observed that pressure increases
reduced pressure (P/Pc), ranging from 1.01 to 1.11 resulted in increased dissolution of metal chlorides in
and from 1.01 to 1.5, respectively, are regarded as SC ethanol, whereas pressure decreases caused dis-
SCFs. In the critical region, the liquid and gaseous solved materials to precipitate as a ‘‘snow.’’ This
phases are indistinguishable, as the substance exhibits increased solubility was later determined to be higher
a liquidlike density and gaslike viscosity and diffusiv- than the solubility predicted by vapor pressure alone.
ity, allowing for good mixing and mass transfer. These During the first half of the 20th century, SCF tech-
characteristics can be manipulated because SCFs show nology attracted little attention. The spark igniting
high rates of changes in the solubility of non-volatile interest came from process improvement technology.
solutes in response to small temperature and/or press- In 1936, Wilson et al. devised a propane deasphalting
ure modifications. process that made use of the increased solvating power
Critical temperatures of substances vary over a wide of propane near critical temperature and pressure con-
range, allowing for SCF selection for specific applica- ditions. This process enabled the selective separation of
tions. Supercritical CO2 is especially useful in the food lube oil feed stocks into paraffin wax, asphalt, heavy
and pharmaceutical industries where toxicity of the ends, naphthenes, and purified light oil.[5,6] Based on
extraction medium, solvent entrapment, and thermal this, the Solexol process was developed several years
stability of materials are concerns. Specifically, SCFs later by utilizing propane at near critical conditions
required for processing thermally labile compounds for the purification and separation of vegetables and
such as proteins and peptides should have a critical fish oils. The purpose of the process was to concentrate
temperature that is close to ambient conditions (e.g., polyunsaturated triglycerides in vegetable oils and to
CO2 and ethylene with critical temperatures of extract vitamin A from fish oils.[6,7]
31.1
C and 9.3
C, respectively). In 1971, Zosel[8] of the Studiengesellschaft Kohle
The purpose of this article is to provide an overview discovered that caffeine could be removed from green
of the applications of SCF technology in pharmaceuti- coffee beans in a reasonable time at moderate tempera-
cal research and product development. It is important tures using CO2 as a supercritical solvent for extrac-
to recognize that SCF technology is by no means a new tion. The coffee brewed from the decaffeinated beans
technique with a history dating back to over a century. produced a consistent flavor as naı̈ve beans and had
Current pharmaceutical applications of SCF tech- the advantage of being ‘‘natural,’’ considering that
nology include: drug extraction and analysis, drug par- there was no use of chlorinated chemical solvents.
ticle and drug polymorph engineering, and preparation These observations led to the application of SCF tech-
of drug delivery systems. This article will address nology for decaffeination of green coffee in the 1970s
advances in these areas as well as scale up issues as the first major commercialized SCF process for
associated with SCF pharmaceutical processes. comestibles.
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-120012011
3568 Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved.
Supercritical Fluid Technology in Pharmaceutical Research 3569

success in pharmaceutical applications of SCFs is the

Bradford Particle Design. The company underwent
restructuring in January 2003 to form Nektar
Therapeutics from a fusion of three entities: Bradford
Particle Design, Inhale, and Shearwater, reflecting a
broader approach to drug delivery.
Currently, the SCF technology has a widespread

Suspensions
utility ranging from food processing to pharmaceutical

Super–
applications. The use of SCFs such as CO2 is proving
to be environmentally benign and economical, with
the added advantage of reduced residual solvents in
food and pharmaceutical products.

FUNDAMENTALS OF SCF TECHNOLOGY

Fig. 1 Pressure–temperature phase diagram of a pure
component indicating the supercritical region. (From SCF Selection
Ref.[61].)
Careful SCF selection is important to reduce proces-
sing time and to improve efficiency. However, the
The first citations on SCF technology applied to choice is usually restricted by the desire of reasonable
active pharmaceutical ingredients are related to pure critical parameter values and costs, chemical inertness,
component precipitation. The goal was to reduce drug low toxicity, and little environmental impact.[9] The
particle size and to enhance a drug’s dissolution rate most frequently used solvent in pharmaceutical proces-
and/or bioavailability.[22–25,31,45–47] Subsequent reports sing is supercritical carbon dioxide (SC CO2), which
addressed the coprecipitation of a drug and a biocom- has low critical temperature (31.1 C) and pressure
patible polymer.[25–36,43] Some authors compared the (73.8 bar), permitting the processing of labile pharma-
characteristics of drugs or composites formed by ceuticals. Table 1 summarizes the most commonly used
supercritical processes to those formed by conven- SCFs for a variety of applications. Being inexpensive,
tional precipitation processes. The effect of process non-toxic, abundant, and environmentally safe, SC
parameters on morphology, particle size, and particle CO2 is an attractive solvent or antisolvent. With small
size distribution were often analyzed. Recent investiga- variations in pressure and temperature, SC CO2 sol-
tions improved traditional SCF processes by enhanc- vent power is tunable. Although CO2 is non-polar with
ing mass transfer and preventing agglomeration.[40–42] a dipole moment of zero, the polarity can be manipu-
The number of patents related to ‘‘SCF’’ and lated by adding small amounts of entraining solvents
‘‘pharmaceutical’’ is in excess of 400 from 1975 to such as alcohols, esters, and ketones.[9] The solvent
the present. Pharmaceutical applications of SCF power, or lack thereof, is an essential aspect in selecting
processing came to fruition during the 1990s. One key an SCF.

Table 1 Properties of commonly used SCFs

Critical Critical Critical density Solubility parameter
Solvent pressure (bar) temperature ( C) (g/mL) (d1), (cal/cm3)0.5a
Ethylene 49.7 9 0.22 1.7
Carbon dioxide 73.8 31.0 0.468 2.0
Nitrous oxide 71.5 36 0.45 2.0
Propylene 46 91.8 0.232
Propane 42.5 96.8 0.217 1.5
Ammonia 111 132 0.24 2.5
n-Pentane 33.7 196.6 0.237
Ethanol 61.4 240.8 0.276
Water 221.2 374.1 0.315 3.5
a
Estimated according to the method of Giddings (1968).
(From Ref.[44].)
 3570 Supercritical Fluid Technology in Pharmaceutical Research

SCF Processes RESS, GAS, and PCA techniques are summarized

subsequently.
Expansion of a supercritical solution containing a RESS is useful for materials that are soluble in CO2;
solute and precipitation of solute dissolved in an however, many polymer, biopolymer, and drug
organic solvent utilizing SCFs as antisolvents are two substances are not CO2-soluble, thereby limiting this
basic techniques of SCF processing for particle pro- application. Cosolvents such as methanol or acetone
duction.[10,11] The most common expansion methods can increase the solvent power of SCFs during RESS.
Suspensions

are rapid expansion of supercritical solutions (RESS) In drug delivery applications, RESS has been used to
Super–

and supercritical fluid nucleation (SFN). Precipitation prepare polymeric films, microparticles, nanoparticles,
methods employing SCFs as antisolvents include: liposomes, and porous foams. A typical RESS appa-
gas antisolvent recrystallization (GAS), supercritical ratus consists of a preheater, an extraction unit, and a
antisolvent recrystallization (SAS), supercritical precipitator (Fig. 2A). After the solute has been placed
antisolvent with enhanced mass transfer (SAS-EM), in the extraction unit, the compressed and preheated
solution-enhanced dispersion by supercritical fluids supercritical solvent is passed through this unit. When
(SEDS), precipitation with compressed antisolvents the solute has dissolved in the SCF, the supercritical
(PCA), and aerosolized solvent extraction system solution is expanded through a capillary or laser-
(ASES). The basic schematic of RESS, GAS, and drilled nozzle into a precipitation unit. Typical nozzles
PCA/SAS/ASES processes are depicted in Fig. 2. have internal diameters between 25 and 150 mm with
The primary requirements for processing with short lengths to prevent precipitation inside the nozzle
due to the rapid pressure drop. Rapid decompression
of the solution in the precipitation unit leads to high
Heated supersaturation and precipitation of desired solids.
A nozzle High supersaturation ratios and rapidly propagating
mechanical perturbations greater than those achieved
by thermal perturbations afford RESS the potential
CO2 Source

Pre-heater

Precipitator

Particles
Extractor

to produce small, uniformly sized particles. Product

contamination is minimal because the solvent is a
dilute gas after expansion and no additional solutes
are needed to initiate precipitation. Pre-expansion
and postexpansion temperature and pressure, nozzle
geometry, and solute properties are among the factors
that determine the size and morphology of delivery
B systems prepared by RESS.
GAS processing uses a dense gas as an antisolvent
for precipitating a solute dissolved in an organic
CO2 Source

Precipitator

solvent (or another SCF).[11,12] Introduction of an anti-

Expanded solvent gas or an SCF into a batch of organic solvents
solution
containing a dissolved solute results in dissolution of
Particles the gas (many gases are at least partially miscible with
organic solvents), lowering the solvent strength of the
liquid phase. On sufficient solvent strength reduction,
the organic solution will no longer be a good solvent
for the solute initiating the onset of nucleation, thus
Solution
inducing precipitation (Fig. 2B). A solute must be
C soluble in an organic solvent or another SCF to be
Nozzle processed by GAS and must be reasonably insoluble
in the antisolvent gas or SCF. Moreover, the solvent
CO2 Source

Solution must be at least partially miscible with the antisolvent.

Precipitator

SC CO2 exhibits weak solvent strength toward

many organic and polymeric solutes. This antisolvent
Particles or non-solvent nature of SCFs can further be
employed by spraying an organic solution of the drug
and/or polymer through a nozzle into a compressed
gas or SCF (Fig. 2C). This process, termed PCA,
Fig. 2 Schematics of (A) RESS; (B) GAS; and (C) PCA/ employs either a liquid or supercritical antisolvent.
SAS/ASES. (From Ref.[44].) When a supercritical antisolvent is used, the process
Supercritical Fluid Technology in Pharmaceutical Research 3571

is often referred to as a SAS process or ASES. GAS, Chromatography

PCA, SAS, and ASES processes offer the flexibility
of solvent choice and higher solute throughput than GC, LC, and SFC chromatography units have been
RESS. For this reason, GAS and related processes used in combination with SFE units. The SFE-LC
are used more often in producing drug delivery techniques aim to separate analytes of high polarity,
systems, ranging from films to nanoparticles. Various molecular weight, or high thermal lability that cannot
nozzle types, including standard capillary nozzles, be addressed via GC or SFC separations. SFE-LC

Suspensions
ultrasonic atomizers, and coaxial nozzles, have been interfacing is the most intricate and difficult due to a

Super–
employed to spray solutions in these processes. combination of a preparative technique that produces
Parameters affecting the control of particle size and gas with a chromatographic technique that handles a
morphology include: operating pressure, temperature, liquid mobile phase. A preferred coupling is SFE-GC
jet breakup, droplet size, and mass transfer rates. The because of the high sensitivity and flexibility of com-
antisolvent addition rate influences the size of the final patible detectors; however, the analyte must be ther-
product; with slow antisolvent addition, there is low mally labile. With SFE-SFC, because the solvent
supersaturation and relatively few nuclei are formed. used to inject the sample is the same as the mobile
Because a large quantity of the solute still remains phase, the basic requirement for effectively coupling
in the solvent, these nuclei grow to large crystals. two techniques (i.e., compatibility between system 1
Rapid antisolvent addition rates produce high super- output and system 2 input) is met. An additional
saturation, and large numbers of nuclei are formed. advantage of this technique over SFE-GC and
With low solute amounts remaining in the solvent, SFE-LC is the reduced probability of insoluble sample
nucleation will not occur to a great extent. components reaching the chromatographic column.[9]

Drug Analysis in Human/Animal

APPLICATIONS IN PHARMACEUTICAL Tissues and Fluids
RESEARCH
A GC mass spectrometry (MS) method following SFE
Drug Extraction and Analysis was used to determine amphetamines in hair.[13] Fol-
lowing sequential washes with sodium dodecyl sulfate
Supercritical fluid extraction (SFE or fractionation) can (SDS), CH2Cl2, methanol, and water, and drying, the
be effectively used for selective removal of components sample was extracted with 90% CO2 and 10%
from a variety of solid, semisolid, and liquid matrices CHCl3/isopropanol at 262 bar and 70
C in the
for subsequent analysis. The matrices encountered in dynamic extraction mode for 30 min. Mephentermine
pharmaceutical applications are complex, and analytes was used as an internal standard. The detection limits
of interest are at trace levels, which SFE can address were 0.02–0.1 ng/mg, relative standard deviations were
with such matrices. SFE can effectively remove undesir- 11–28%, and recoveries were 71–84%.
able components from pharmaceutical products, and Huopalahti and Henion[14] reported anabolic ster-
various chemicals extracted with SFE can be impreg- oid extraction from bovine tissues. SFE was performed
nated into other matrices as well. There are two modes with CO2 at 60
C and 405 bar, with analytes collected
in which extraction and analysis can be accomplished: in precooled methanol and quantitated using LC-MS
off-line, in which extracted analytes are collected in a analysis. Residues of 2-thiouracil, 6-methyl-2-thiouracil,
device independent of the measurement instrument, 6-propyl-2-thiouracil, and 6-phenyl-2-thiouracil in bovine
and on-line, in which the extraction step is combined muscle tissues were extracted with SC CO2 and analyzed
with a separation technique [gas chromatography using GC-MS.[15]
(GC), liquid chromatography (LC), or SCF chromato- Temazepam[16] and morphine[17] have been sepa-
graphy], followed by drug quantitation. In addition to rated from a whole blood matrix using SC CO2. In
these modes of extraction and analysis, there are two the first instance, the extractants were CO2 and ethyl
modes for extraction purposes: static and dynamic. acetate at 65
C and 207 bar, and recoveries ranged
For static mode, samples are allowed to steep in the from 80% to 100%. The extraction was monitored at
extractant, whereas in dynamic mode, the extractant 254 nm, with individual separation of the analytes car-
continuously flows through the extraction vessel. ried out by high-performance liquid chromatography
Both modes of extraction encounter problems such as (HPLC). For morphine extraction from dried blood,
restrictor plugging (dynamic) and longer extraction methanol/triethylamine was added as a modifier to
times (static). To achieve optimal separation and 90% CO2 at 100
C and 241 bar, and the extracts were
analysis, a combination of the two modes is often analyzed by GC-MS. The SFE method proved to be
used.[9] faster and cleaner than solid-phase extraction (SPE).
 3572 Supercritical Fluid Technology in Pharmaceutical Research

Drug Analysis in Pharmaceutical Preparations dissolution rate, and bioavailability. In recent years,
crystal and particle engineering of pharmaceutical
SFE has been used to extract active components from materials and drug delivery systems with SCF tech-
tablets. Methanol was used as the modifier of CO2 for nology has gained momentum due to the limitations
the extraction of benzodiazepines from tablets or of conventional crystallization, comminution (crush-
capsules in static and dynamic modes for 5 and ing, grinding, and milling), and spray drying
10 min, respectively.[18] The SFE conditions were set approaches. Conventional recrystallization using
Suspensions

at 65
C and 101 bar, and the drugs were analyzed using liquid antisolvents requires extensive use of organic
Super–

GC-MS analysis. solvents. Mechanical forces during comminution may

An interesting SFE application consists of assay degrade some pharmaceuticals, and spray drying
development for quantitation of the active enantiomer may cause thermal stress and degradation of some
of misoprostol in a controlled-release drug formu- products. Table 2 shows a comparison of these
lation.[19] SC CO2 modified with 5% formic acid at conventional techniques with that of SCF processing.
75
C and 334 bar was effective in cleaving the covalent To further demonstrate the controllability of particle
linkage between misoprostol and the polymer, and morphology and associated differences between con-
subsequent extraction. With this approach, high drug ventional crystallization and SCF processing, scanning
recoveries were observed over a short extraction per- electron micrographs of both freeze-dried and SEDS-
iod. Static/dynamic SFE with CO2 and 8.7% methanol processed lysozymes are shown in Fig. 3.[21]
modifier gave optimal extractions from sustained- Fusaro and Mazzotti[22] demonstrated the versa-
release tablets.[20] The average recovery was 98.6%, tility of the GAS process in controlling paracetamol
and this SFE method used 80% less solvent than the precipitation. The average particle size of monoclinic
traditional liquid extraction procedure. paracetamol crystals formed ranged from 50 to
250 mm, with the particle size decreasing as the CO2
antisolvent addition rate is increased. Additionally,
Preparation of Drug Powders and Polymorphs the study addressed the robustness of the precipitation
procedure by quasi-scaling. Crystals produced using
The demand for small, narrowly distributed particles 400-mL and 1-L precipitation vessels were qualitatively
with well-defined polymorphic form is critical in drug and quantitatively in agreement. Table 3 summarizes
delivery applications, considering the direct role that the use of various SCF techniques for controlled
particle size and polymorphism play in solubility, crystallization of plain drug particles.

Table 2 Comparison of different particle formation techniques

Factors Micronization (milling) Spray drying SCF precipitation
Process Secondary Direct Direct
Typical product Relatively dense powders Low-density powders Low-density powders
description formed by irregularly formed by composite formed by loosely packed,
shaped, rough-surface porous or hollow regularly shaped
crystalline particles amorphous particles crystalline particles
Major advantage area Established, proven Good particle aerodynamics Pure non-cohesive powders
Typical composition Pure small molecules Biological or small active Pure small molecules or
molecules with excipients composites with polymers
Potential for Low High High
coformulation
Control over Adequate for small molecules; Poor for small molecules; Excellent for small molecules;
physical stability poor for biomolecules adequate for biomolecules adequate for biomolecules
Batch consistency Poor Adequate Good
Particle size control Adequate High Good
FPF target 30% 80% 60%
Current industrial scale Commercial Small manufacturing Pilot
Current cost Low Moderate More expensive than other
two technologies
Potential for development Low Moderate High
(From Ref.[38].)
Supercritical Fluid Technology in Pharmaceutical Research 3573

SCF technology can now be claimed as useful in

producing particles ranging from 5 to 2000 nm.[25] This
patent covers a process that rapidly expands a solution
of the compound and phospholipid surface modifiers
in a liquefied gas into an aqueous medium, which con-
tains the phospholipids. Expanding into an aqueous
medium prevents particle agglomeration and particle

Suspensions
growth, thereby producing particles of a narrow size

Super–
distribution.

Drug and Delivery System Design

Fig. 3 Scanning electron microscopy (SEM) of (A) a freeze- Microparticles (RESS)

dried lysozyme sample and (B) the same sample prepared
via SEDS. (From Ref.[21].) SCF preparations of various drug and polymeric
microparticles have utilized both RESS and assorted
antisolvent techniques. Composite microparticles can
be formed in fewer steps using SCF processes, as
Aiming to increase dissolution rate and bio- shown in Fig. 4. Tom and Debenedetti[26] used a RESS
availability, particles of the highly insoluble calcium process to produce DL-poly(lactic acid) (DL-PLA) parti-
channel blockers, nifedipine and felodipine, and the cles of lovastatin, a cholesterol-reducing drug. Particles
hypolipidemic agent, fenofibrate, were micronized by containing embedded lovastatin needles were formed
precipitation from gas-saturated solutions (PGSS) and were unsuitable for drug delivery, but this was
technique.[23] These various preparations were sprayed the first study to explore the coprecipitation of a drug
into the compressed gas through a calibrated nozzle, and a polymer from a supercritical solvent. The RESS
with orifice sizes ranging from 250 to 400 mm. Depen- process has also been used to coprecipitate naproxen
dent on the pre-expansion conditions, 15–30-mm with L-poly(lactic acid) (L-PLA) from SC CO2.[27] In
nifedipine particles and 42-mm felodipine particles were this study, SC CO2 was passed through an extraction
obtained. The fenofibrate processed at 70
C and 190 vessel containing a mixture of naproxen and L-PLA,
bar produced particles from 7 to 32 mm, and these and then expanded through a 50-mm capillary tube at
larger particles (larger than native samples) occurred 190 bar and a pre-expansion temperature of 114
C.
due to agglomeration. The particle morphology was spherical with diameters
Kordikowski, Shekunov, and York[24] utilized liquid ranging from 20 to 90 mm. Table 4 summarizes various
and SC CO2 in the SEDS process to control the drug carrier particles prepared using different SCF
polymorphic forms of sulfathiazole. The investigation processing techniques.
studied effects of temperature and CO2/solvent flow In addition to low-molecular weight drugs, modified
rates on generating pure polymorphs of the four enan- RESS processes are capable of encapsulating macro-
tiotropic forms. Sulfathiazole was initially saturated molecular drugs such as proteins and peptides.
in acetone and methanol at 1.5% and 1.0% wt/vol, Mishima et al.[28] reported the microencapsulation of
respectively. Both saturated solutions were processed lipase and lysozyme in polymeric particles. The parti-
at 200 bar over a temperature range of 0–120
C. The cles were prepared using polymers such as PEG 4000,
acetone flow rate 0.2–12.8 mL/min per 10 mL/min PEG 6000, PEG 20,000, poly(methyl methacrylate),
CO2 was varied to attain a mole fraction of acetone poly(DL-lactic acid), poly(DL-lactide-co-glycolide), and
of 0.01–0.42. Variations in temperature and compo- PEG poly(propylene glycol)–PPG–PEG, a triblock
sition produced changes in polymorphic form, attain- copolymer. The solubility of these polymers in CO2
ing only amorphous form and form I. This indicated was enhanced by using low-molecular-weight alcohols
that crystallization from acetone was kinetically con- (methanol, ethanol, and isopropanol) as cosolvents. In
trolled. From the saturated methanol solution, the flow this process, known amounts of polymer, protein,
rate 0.2–25.6 mL/min per 10 mL/min CO2 was varied and cosolvent were placed in the high-pressure vessel
to produce a mole fraction of methanol of 0.02–0.72. containing SC CO2. This mixture was stirred by an
By selecting an appropriate temperature (at all flow agitator rotating at 200 rpm for approximately 4 hr, and
rates), three pure forms of sulfathiazole were crystal- then kept for more than 1 hr without agitation. The
lized: form I, form III, and form IV. Methanol as a dissolved polymer with dispersed protein preparation
hydrogen bond donor possessed a greater ability to was sprayed through a capillary nozzle toward a target
stabilize different forms of sulfathiazole than acetone. plate. Parameters such as the thickness of the polymer
 Suspensions
Super–
3574

Table 3 Plain drug particles prepared using SCF processes

Flow rate

(b) Drug Temperature/

Process/ Nozzle solution pressure Mean
Drug solvent id (lm) (a) CO2 (mL/min) (
C/bar) size (lm) References
Paracetamol GAS/acetone (a) 0.067–6 5–40/– 50–250 [22]
aggregated
Nifedipine PGSS 400/250 165,175, and 15–30 [23]
185/100–200
Insulin SAS/DMSO 30/50 (a) 9–26 L/min (b) 0.9–1.7 28–46/90.6–14.2 [47]
Lysozyme SAS/DMSO 30/50 (a) 10–21 (b) 0.2–2.4 27–45/91–142 1–5 [47]
Trypsin SAS/DMSO 30/50 (a) 5–20 L/min (b) 0.5–2.1 27–47/73–136 1–5 [47]
Insulin GAS/DMSO 30 (a) 8 L/min (b) 0.3 25 and 35/86.2 2–3 [59]
Salmeterol PCA (a) 8 (b) 0.1–0.3 35/100 10–16 [60]
Xinafoate 35/200 9–12
35/300 4–7
Budesonide PCA 100 (a) 10–50 (b) 1.4–2.7 35 and 40/78.9–139 1–2 [31]

PGSS ¼ particles from gas-saturated solution (gaseous CO2 is dissolved in a melted drug and the gas-saturated solution is expanded).
Supercritical Fluid Technology in Pharmaceutical Research
Supercritical Fluid Technology in Pharmaceutical Research 3575

(A) Emulsion solvent evaporation (B) Supercritical fluid respectively (vs. 10% theoretical for both). These were
method process processed at room temperature and flowing CO2
Aqueous phase Organic phase antisolvent at 69.0 bar through a 100-mm nozzle.
PVA solution Polymer + Drug Drug + Polymer Martin et al.[31] precipitated budesonide and
(2% w/v) + Solvent in SCF or organic budesonide–PLA microparticles by a PCA technique.
solvent Utilizing SC CO2 as an antisolvent at various tempera-
Mixing & tures (35
C and 40
C) and pressures (78.9, 85.8, 92.7,

Suspensions
sonication
and 138 bar), the drug solution and antisolvent were

Super–
While stirring, add dropwise Expansion or spraying passed coaxially, as drug solution was introduced
to excess aqueous phase into antisolvent through a 100-mm capillary tube. Typical experiments
and stir overnight involved two 20þ sec sprays of budesonide–PLA
solution at 1.4 mL/min, followed by CO2 equivalent
Centrifuge to obtain Microparticles volume purging to remove the residual solvent (methylene
microparticles
chloride). The mean diameters of processed budeso-
nide and budesonide–PLA particles were 1–2 mm. The
Wash particles (3x) with
DI H2O and reconstitute
budesonide–PLA microparticles had a loading of
in DI H2O 7.94%, which was equivalent to an 80% encapsulation
efficiency. The in vitro release studies indicated a 50%
Lyophilize reconstituted budesonide release from the microparticles at the end of
product to obtain 28 days. The unprocessed budesonide, SCF-processed
dry powder budesonide, and SCF-processed budesonide–PLA parti-
cles are shown in Fig. 5.
Microparticles
The aerosol solvent extraction system has been used
by investigators to prepare microparticles.[32,33] Bleich
et al. tested a variety of biodegradable polymers
Fig. 4 Preparation of composite microparticles using con- including L-PLA, DL-PLA, DL-PLGA (lactide/glycolide
ventional and SCF approaches. 50 : 50 and 75 : 25), and poly(n-hydroxybutyric acid)
(PHB) for their suitability to be processed by an ASES
process utilizing SC CO2. It was found that both
coating the protein, particle size, morphology, and par- DL-PLA and DL-PLGA (75 : 25) were extracted by SC
ticle size distribution were controlled by changing the CO2. DL-PLGA (50 : 50) formed a film at the bottom
feed composition of the polymer. For instance, increas- of the collecting vessel. Both semicrystalline polymers
ing the feed concentration of PEG 6000 from 10 to DL-PLA and PHB formed spherical microparticles with
20 wt.% resulted in a particle size increase from 20 to mean sizes between 1 and 10 mm.[34]
70 mm. However, no information regarding protein Ghaderi et al.[35] used SEDS to produce microparti-
encapsulation efficiency was provided. cles of L-PLA, DL-PLG, DL-PLA, and PCL. Solutions
of these polymers in different organic solvents were
Microparticles (antisolvent processes) sprayed into SC CO2. The size and morphology of
the particles formed strongly depended on: solvent
Most drugs do not exhibit good solubility in SC CO2; type, polymer concentration, temperature, pressure,
therefore, large numbers of studies employed CO2 as and density of the antisolvent gas. The experiment
an antisolvent. Mueller and Fisher[29] first patented a for DL-PLGA used a mixture of acetone, ethyl acetate,
process that used an antisolvent SCF process to pro- and isopropanol in a ratio of 4 : 5.6 : 0.4. Addition of
duce polymeric drug microparticles of clonidine hydro- isopropanol in small amounts, a poor solvent for DL-
chloride encapsulated by DL-PLA. The polymer/drug PLGA, helped attain higher degrees of supersatura-
solution was dissolved in methylene chloride and tion. This agent aided in the production of discrete
sprayed into a vessel containing SC CO2. Bodmeier amorphous microparticles of this polymer. L-PLA
et al.[30] prepared polymeric drug-loaded microparti- formed discrete microparticles with mean diameters
cles by atomizing organic polymer/drug solutions by from 0.5 to 5.3 mm under various processing conditions
the PCA process. Low polymer concentrations (1%), (temperature and pressure of 35–40
C and 130–185 bar,
high CO2 flow rate and density, and low temperatures respectively). The semicrystalline polymer L-PLA was
(below the glass transition temperature Tg of the more easily processed by the SEDS technique, showing
polymer) favored the formation of small-diameter, size distributions sensitive to CO2 density alterations.
narrowly distributed L-PLA microparticles. PLA The particle size increased (0.5–5.3 mm) as the CO2
encapsulated chloropheniramine maleate and indo- density (0.85–0.69 g/mL) decreased. The amorphous
methacin with drug loadings of 3.73% and 0.73%, polymers DL-PLA and PCL also formed microparticles
 Suspensions
Super–

Table 4 Drug–polymer composite particles prepared using SCF processes

3576

Flow rate

(b) Drug Temperature/

Nozzle solution pressure
Polymer/drug Process/solvent id (lm) (a) CO2 (mL/min) (
C/bar) Morphology References
a
L-PLA/naproxen RESS/SC CO2 50 60/187–210 [27]
90–140b 29–40c
PEG/lysozyme or lipase RESS/EtOH 280 (a) 20 (b) 1 35/200 [28]
PMMA/lysozyme or lipase RESS/EtOH 280 (a) 20 (b) 1 35/200 [28]
PLA/lysozyme or lipase RESS/EtOH 280 (a) 20 (b) 1 35/200 [28]
PLGA/lysozyme or lipase RESS/EtOH 280 (a) 20 (b) 1 35/200 [28]
PEG–PPG–PEG triblock RESS/EtOH 280 (a) 20 (b) 1 35/200 [28]
copolymer/lysozyme or lipase
L-PLA/Clorpheneniramine PCA/CH2Cl2 100 25/69 and [30]
maleate (10/3.73)d agglomerated
L-PLA/indomethacin (10/0.73)d PCA/CH2Cl2 100 25/69 [30]
e
PLA/Naltrexone PCA/CH2Cl2 1000f (a) 20 (b) 1 35–38/85–90 [43]
(2, 5, 10/1.7, 6.1, 9.2)d
PLA/gentamicine PCA/CH2Cl2 1000f (a) 20 (b) 1 35–38/85–90 [43]
(5, 10, 15, 20/4.2, 12.2, 17.4, 24.7)d
PLA/rifampin (5–50/0.5–37.4)d PCA/CH2Cl2 1000f (a) 20 (b) 1 35–38/85–90 [43]
PLA/Budesonide PCA/CHCl3 100 (a) 10–50 (b) 1.4–4.7 35 and 40/78.9–139 and [31]

agglomerated
PLGA/estriol (100/110)d ASES/CH3OH þ — (a) 11 kg/hr (b) 6 34/100 and [33]
TFE þ CH2Cl2 agglomerated
PLGA/BSA (100/114.15)d ASES/CH3OH þ — (a) 11 kg/hr (b) 6 34/100 and [33]
TFE þ CH2Cl2 agglomerated
Triblock polymerg/estriol ASES/CH3OH þ — (a) 11 kg/hr (b) 6 34/100 and [33]
(100/103.2)d TFE þ CH2Cl2 agglomerated
Triblock polymerg/BSA ASES/CH3OH þ — (a) 11 kg/hr (b) 6 34/100 and [33]
(100/105.6)d TFE þ CH2Cl2 agglomerated
BSA ¼ bovine serum albumin; PLA ¼ polylactic acid; PLGA ¼ poly(lactic-co-glycolic acid); PEG ¼ poly(ethylene glycol); PPG ¼ poly(propylen -trifluoroethanol.
a
Extraction temperature/pressure.
b
Pre-expansion temperature.
c
Expansion temperature.
d
Percent theoretical loading/experimental loading.
e
Dissolved in dichloromethane by hydrophobic ion pairing with AOT.
f
Vibrating nozzle at 120 kHz.
Supercritical Fluid Technology in Pharmaceutical Research

g
b-Poly-L-lactide–co-D,L-lactide–co-glycolide (62.5 : 12.5 : 25).
Supercritical Fluid Technology in Pharmaceutical Research 3577

CO2

Small non-porous SC SC CO2 penetrates

particles conditions particles

Suspensions
Super–
Depressurization SC CO2 and particle
expansion

Rapid escape
of CO2

Large porous
particles

Fig. 6 Mechanism of large porous polymeric particle forma-

tion by pressure quench method using SC CO2. (From
Fig. 5 Above: SEM of (A) unprocessed and (B) SCF- Ref.[36].)
processed budesonide particles at 40
C and 85.8 bar. Below:
SEM of budesonide–PLA particles processed at 40
C and
85.8 bar, 7.94% wt/wt budesonide. (From Ref.[31].) for pulmonary delivery. Furthermore, large porous
deslorelin–PLGA microparticles better sustained pul-
monary drug delivery compared with conventional
particles.[37]
with mean diameters ranging from 105 to 216 mm and There are definite advantages in producing powders
from 27 to 212 mm, respectively. for respiratory formulations using SCF processes.[38]
These include: enhanced dispersibility at low airflow
Porous microparticles rates or aerodynamic turbulent stress, reduced
adhesive and cohesive interactions due to relatively
SCFs have been utilized in the formation of large low surface energy, and the possibility of using drug
porous deslorelin–PLGA microparticles.[36] Following powders with carriers or excipients. Dry powder
the preparation of deslorelin–PLGA (50 : 50) micro- inhalers (DPIs) require relatively low dispersion forces
particles by a conventional emulsion–solvent evapora- to deaggregate powders. SCF-processed materials
tion method, these particles were exposed to SC provide a more consistent crystalline form and low dis-
CO2 for discrete time intervals followed by rapid persion forces, providing a potential means to deliver
isothermal depressurization. A schematic of the large respiratory drugs independent of the specific geometry
porous polymeric particle formation mechanism is or flow rate of a given DPI.[38]
depicted in Fig. 6. Exposure of SC CO2 (1200 psi and
33
C for 30 min) increased the particle size from 2.2 Microporous foams
to 13.8 mm and average pore diameter from 90 to
190 nm. Pores were visible as irregular surfaces, as seen Hile et al.[39] prepared porous PLGA foams using an
in Fig. 7. After SC CO2 treatment, residual methylene SCF technique capable of releasing an angiogenic
chloride levels were undetectable (<25 ppm), which agent, basic fibroblast growth factor (bFGF), for tissue
was drastically different from untreated microparticles engineering applications. In this technique, a hom-
(4500 ppm) and much less than the United States Phar- ogenous water-in-oil emulsion consisting of an aque-
macopeia (USP) limit (500 ppm). The drug loading was ous protein phase and an organic polymer solution
5.16  0.93%, which was not significantly different was prepared first. This emulsion was filled in a longi-
from the conventionally prepared particles, indicating tudinally sectioned and easily separable stainless steel
that no drug was lost during the SC CO2 processing. mold. The mold was then placed into a pressure cell
The large porous particles exhibited reduced macro- and pressurized with CO2 at 80 bar and 35
C. The
phage uptake, indicating their potential application pressure was maintained for 24 hr to saturate the
 3578 Supercritical Fluid Technology in Pharmaceutical Research
Suspensions
Super–

Fig. 7 SEM of deslorelin–PLGA microparticles (A) before and (B) and (C) after SC CO2 treatment at 33
C and 1200 psi for
30 min. (From Ref.[36].)

polymer with CO2 for the extraction of methylene exhibiting linear release kinetics over a 7-week period.
chloride. Finally, the setup was depressurized for By employing hydrophobic ion pairing, this study
10–12 sec, creating a microporous foam that sustained applied SCF processing to ionic pharmaceuticals.
the release of the growth factor.
Macromolecule powders
Nanoparticles
Operating conditions with SC CO2 are considered
Chattopadhyay and Gupta achieved significant benign, especially for processing macromolecules such
improvement in an antisolvent technique using a as peptides, proteins, and nucleic acids. The conven-
SAS-EM process, yielding particles with narrow size tional method of protein production lyophilization
distributions and mean sizes up to 10-fold smaller. In subjects macromolecules to freeze–thaw cycles, poten-
SAS-EM, the solution jet is deflected by an ultrasonic tially affecting solution conformation and biological
vibrating surface at a frequency that atomizes the jet. activity.[44]
The ultrasound field generated by the vibrating surface a-Chymotrypsin was precipitated from both com-
enhances mass transfer and, by promoting mixing, pre- pressed CO2 and a liquid fluorinated antisolvent,
vents agglomeration. Particle size is easily controlled hydrofluoroether (HFE). Made in an acidic environ-
by adjusting the vibration intensity of the deflecting ment to inhibit self-proteolysis, the a-chymotrypsin
surface through the variable power supply of the solutions at a concentration of 12 mg/mL were intro-
ultrasound transducer. This technique demonstrated duced via a 100-mm-diameter capillary nozzle into
nanoparticle formation of different pharmaceuticals flowing CO2/ethanol (2.4 : 1 molar ratio) antisolvent
such as lysozyme, tetracycline, and griseofulvin.[40–42] using a PCA technique. Operating conditions of the
Falk et al.[43] used a PCA technique employing an SCF antisolvent were 36
C and 136 bar. The protein
ultrasonic spray nozzle that vibrates at 120 kHz to pro- precipitates were spherical with some agglomeration.
duce nanoparticles of gentamicin, naltrexone, and Use of HFE as an antisolvent produced monodisperse
rifampin with L-PLA. The drug/polymer particle mor- particles that retained protein activity.[45]
phology was spherical, ranging from 200 to 1000 nm in Debenedetti et al. used an antisolvent method to
diameter. The experiment used hydrophobic ion pair- form microparticles of insulin and catalase. Protein
ing to solubilize gentamicin and naltrexone in methy- solutions in hydroethanolic mixture (20 : 80) were
lene chloride. In this process, the polar counter ions allowed to enter a chamber concurrently with SC
(sulfate and chloride) of the ionic pharmaceutical agent CO2.[46] The SCF expanded and entrained the liquid
were replaced with a hydrophobic one (sodium bis-2- solvent, precipitating submicron protein particles.
ethyl-hexyl sulfosuccinate). The more lipophilic rifam- Supercritical antisolvent processing methods also
pin was not solubilized by hydrophobic ion pairing and expose proteins to potentially denaturing environments,
produced rifampin particles with low loading efficiency including organic and supercritical non-aqueous sol-
(0.5–37.4% experimental vs. 5–50% theoretical). How- vents, high pressure, and shearing forces, which can
ever, gentamicin had a high loading efficiency (4.3, unfold proteins such as insulin, lysozyme, and trypsin
12.2, 17.4, and 24.75% wt/wt measured values vs. 5, 10, to various degrees.[47] This led to the development of a
15, and 20% wt/wt theoretical values, respectively) and method, wherein the use of organic solvents is com-
the prepared particles showed low burst release while pletely eliminated to fully obtain active insulin particles
Supercritical Fluid Technology in Pharmaceutical Research 3579

of 1.5–500 mm.[48] In this application, insulin was allowed Frederiksen et al.[54] developed a laboratory-scale
to equilibrate with SC CO2 for a predetermined time, method for the preparation of small liposomes by
and the contents were decompressed rapidly through a encapsulating a solution of aqueous soluble fluorescein
nozzle to obtain insulin powder. Plasmid DNA particles isothiocyanate (FITC) dextran using SC CO2 as a sol-
can also be prepared using SCFs.[49] An aqueous buffer vent. This system, comprised of two main sections,
(pH 8) solution of 6.9 kb plasmid DNA and mannitol contained a high-pressure unit where 1-palmitoyl-2-
was dispersed in supercritical CO2 and a polar organic oleoylphosphatidylcholine (POPC) and cholesterol

Suspensions
solvent using a three-channeled coaxial nozzle. The were dissolved in SC CO2. The second section, a low-

Super–
organic solvent acts as a precipitating agent and as a pressure unit, expanded and mixed the homogenous
modifier, enabling non-polar CO2 to remove the water. SCF solution and aqueous solution containing the
High dispersion in the jet at the nozzle outlet facilitated marker, yielding liposomes. In comparison to the etha-
the rapid formation of dry particles of small size. The nol injection method of liposomal formation, the SCF
plasmid DNA recovered 80% of its original supercoiled method required 15 times less organic solvent to get
state on reconstitution in water. the same encapsulation efficiency. The length and inner
diameter of the encapsulation capillary influenced the
Inclusion complexes encapsulation volume, encapsulation efficiency, and
average size of the liposomes. Thus the critical param-
Enhancing the solubility of drugs such as piroxicam eter for scaleup considerations of this SCF liposome
can be achieved in the formation of inclusion com- system is the encapsulation capillary dimension.
plexes with cyclodextrins. Previous preparations of Preparations of liposomes via SCF processing are
non-polar drug inclusion complexes involved the use designated as critical fluid liposomes (CFLs). CFLs
of organic solvents, producing a large residual solvent have successfully encapsulated hydrophobic drugs
concentration in these inclusion complexes.[50] Cyclo- such as taxoids, camptothecins, doxorubicin, vincris-
dextrins have also been used for the entrapment of tine, and cisplatin.[55] In addition, stable paclitaxel
volatile aromatic compounds after supercritical extrac- liposomes with a size of 150–250 nm were obtained.
tion.[51] Van Hees et al.[52] employed SCF-aided pro- Aphios Co.’s patent (US Patent 5,776,486) on Super-
duction of piroxicam and b-cyclodextrin inclusion FluidsTM CFL describes a method that is useful for
complexes by exposing the physical mixture of the nanoencapsulation of paclitaxel and campothecin in
piroxicam-b-cyclodextrin (1 : 2.5 mol/mol) to SC CO2. aqueous liposomal formulations called TaxosomesTM
The optimum complex yield of 98.5% occurred after and CamposomesTM, respectively.
6 hr of contact with SC CO2 at 15 MPa and 150
C.
On static mode operation, the depressurization of this Polymer impregnation
mixture occurred within 15 sec into a collection car-
tridge, where the contents were ground and homoge- SC CO2 used as a carrier of drug molecules into a poly-
nized. The influence of temperature, pressure, and SC mer matrix has a number of advantages such as the
CO2 exposure time to the complex was studied, and plasticizing ability of CO2 (based on specific interac-
it was determined that temperature and exposure time tions between CO2 and polymer moieties), which both
predominantly governed the inclusion complex yield. enhances the diffusion rates of drug molecules into the
An increase in yields from 1.39% to 94% occurred polymer and facilitates solvent removal. Polymer plas-
with a corresponding temperature increase from 50
C ticization is accompanied by the swelling of the poly-
to 150
C at a constant pressure of 45 MPa. Using an mer matrix, with a concomitant increase in the free
organic, solvent-free, single-step SCF process, Bandi[53] volume of the polymer. Moreover, SC CO2 can reduce
successfully formed budesonide–HPbCB inclusion the melting temperature of semicrystalline polymers.
complexes. These effects are crucial to the impregnation and modi-
fication of polymeric materials.
Liposomes Sheridan et al.[56] incorporated vascular endothelial
growth factor (VEGF) into three-dimensional porous
Liposomes are lipid bilayer vesicles that are artificially matrices of bioabsorbable materials without the use
created to transport biological molecules and other of organic solvents or high temperatures. This gas
therapeutic agents. The requirement for large amounts foaming process combined copolymers of lactide and
of organic solvents during production and the low glycolide (PLGA 50 : 50, 75 : 25, and 85 : 15) with lyo-
encapsulation efficiency of drug products are among philized VEGF and exposed the mixture to CO2 at
the limitations of liposomal production. These pro- 850 psi and 25
C for 24 hr followed by depressurization
blems can be overcome by using SCF processing, at 340 psi/min. The VEGF incorporation efficiency
utilizing SC CO2 as a cosolvent for viable liposome was 90% and 72% for the 85 : 15 and 75 : 25 PLGA,
formation. respectively. The release profiles for 85 : 15 PLGA
 3580 Supercritical Fluid Technology in Pharmaceutical Research

(84% released within the first 2 days) and 75 : 25 (60% product fabrication is among the many advantages
sustained release for 70 days) were quite different, associated with SCF technology. This technology
which was attributed to copolymer intrinsic viscosity should be considered a tool, rather than a panacea,
and porosity changes. Importantly, bioactivity studies in attempts to provide solutions for areas discussed
demonstrated that incorporated and released growth within the article.
factor retained more than 90% of its activity for the
75 : 25 PLGA. These polymeric scaffolds show great
Suspensions

promise in engineering new tissues and promoting

Super–

the vascularization of developing tissues. ACKNOWLEDGMENTS

Polymer fractionation This work was supported by NIH grants DK 064172

and EY 013842.
Current polymeric drug delivery applications employ
polymers that contain a mixture of molecular weight
fractions. SCF technology can provide a means to
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CONCLUSIONS
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Surfactants in Pharmaceutical Products and Systems
Owen I. Corrigan
Trinity College, University of Dublin, Dublin, Ireland

Anne Marie Healy

Suspensions
Department of Pharmaceutics, School of Pharmacy, University of Dublin, Dublin, Ireland

Super–
INTRODUCTION against the inward pull. The work required to increase
the surface area by unit area is termed the surface free
Surface-active agents (surfactants) are substances energy.
which, at low concentrations, adsorb onto the surfaces At the interface between two condensed phases, the
or interfaces of a system and alter the surface or inter- dissimilar molecules in the adjacent layers facing each
facial free energy and the surface or interfacial tension. other have potential energies greater than those of
Surface-active agents have a characteristic structure, similar molecules in the respective bulk phases. This
possessing both polar (hydrophilic) and non-polar is due to the fact that cohesive forces between like
(hydrophobic) regions in the same molecule. Thus molecules tend to be stronger than adhesive forces
surfactants are said to be amphipathic in nature. The between dissimilar molecules. Thus the interfacial ten-
wide range of uses for surfactants in pharmaceutical sion is the force per unit length existing at the interface
products and systems is the subject of this article. between two immiscible or partially miscible con-
densed phases and the interfacial free energy is the
work required to increase the interface by unit area.
PHYSICOCHEMICAL BACKGROUND
Adsorption Phenomena
Surface and Interfacial Tension; Surface
and Interfacial Free Energy Adsorption may be defined as the process of enrich-
ment of one or more substances at a surface[1] or as
Atoms and molecules at surfaces and interfaces possess the taking up of one substance at the surface of
energies significantly different from those of the same another.[2] It can occur at any type of interface. How-
species in the bulk phase. The term ‘‘surface’’ is usually ever, in the context of pharmaceutical systems the
reserved for the region between a condensed phase interfaces where surfactant adsorption is important
(liquid or solid) and a gas phase or vacuum, while are the gas–liquid, liquid–liquid, gas–solid, and
the term ‘‘interface’’ is normally applied to the region liquid–solid interfaces.
between two condensed phases.
In the case of a liquid–gas interface, molecules of Adsorption at liquid–liquid and
the liquid in the boundary can only develop attractive liquid–gas interfaces
cohesive forces with molecules situated below and
adjacent to them. They can develop attractive adhesive Considering a system of two immiscible phases (e.g.,
forces with molecules of the gaseous phase. However at heptane and water), a surface-active molecule that is
the gas–liquid interface, these adhesive forces are quite adsorbed at the interface between the two liquids will
small. The net effect is that molecules at the surface of tend to orient itself with its hydrophilic end toward
the liquid have potential energies greater than those of the more polar liquid (water), and its hydrophobic
similar molecules in the interior of the liquid and end toward the less polar liquid (heptane). Thus the
experience an inward force toward the bulk of liquid. surfactant molecules replace water and/or heptane
This force pulls the molecules of the interface together molecules of the original interface. The interaction
and the surface contracts. across the interface is then between the hydrophilic
Thus, the surface of a liquid behaves as if it were in group of the surfactant and the water molecules on
a state of tension—the surface tension (g)—due to the one side of the interface, and between the hydrophobic
contracting force acting in all directions in the plane of group of surfactant and heptane on the other side of
the surface. the interface. These interactions are much stronger
In order to extend the surface of a liquid it is neces- than the original interactions between the unlike mole-
sary to bring molecules from the interior to the surface cules of heptane and water; therefore the interfacial
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-100200018
Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved. 3583
 3584 Surfactants in Pharmaceutical Products and Systems

tension is significantly reduced by the adsorption of 4. Adsorption by polarization of p electrons,

surfactant at the interface (i.e., the inward pull for each where the adsorbate contains electron-rich aro-
phase at the interface is reduced). matic nuclei and the adsorbent has strongly
Air consists of molecules that are mainly non-polar. positive sites
Surface tension reduction by surfactants at the air– 5. Adsorption by dispersion forces, i.e., London–
aqueous interface occurs due to adsorption of surfactants van der Waals dispersion forces acting between
at the interface, with the hydrophilic end of the surfactant adsorbate and adsorbent
Suspensions

oriented toward the liquid. The presence of the sur- 6. Hydrophobic bonding.
Super–

factant molecules reduces the net inward pull toward

the bulk liquid, and therefore reduces the surface tension.
The effect of a surfactant on the lowering of surface
Contact Angles and the Wetting of Solids
tension is shown in Fig. 1. The surface tension is low-
ered even at low concentrations of surfactant. As the
A drop of liquid when placed on a flat, homogeneous
surfactant concentration is increased, the surface layer
solid surface, comes to equilibrium, assuming a shape
becomes saturated with surfactant molecules, and
which minimizes the total free energy of the system.
micelles form within the bulk liquid as an alternative
The angle between the liquid and the solid is called
way of shielding the hydrophobic portions of the sur-
the contact angle (y), the angle being measured
factants from the aqueous environment; the surface
through the liquid (Fig. 2). The contact angle may be
tension tends to a constant value. Micelles are small
calculated if the surface and interfacial tensions are
aggregates of surfactant in which the surfactant mole-
known from Young’s equation given in Eq. (1) or (2).
cules are arranged in such a way that the hydrophobic
ends are shielded from the surrounding aqueous
gSA ¼ gSL þ gLA cos y ð1Þ
environment. The concentration at which micelles first
appear in solution is termed the critical micelle concen-
or
tration (CMC).
gSA
gSL
cos y ¼ ð2Þ
Adsorption at solid–liquid interfaces gLA

Adsorption of surfactant from an aqueous solution where gLA is the surface tension of the liquid, gSL is the
onto a solid surface may involve specific chemical interfacial tension existing between the solid and liquid
interaction between the surfactant (adsorbate) and phases, and gSA is the surface tension (or surface free
the surface (adsorbent). energy) of the solid. If y < 90 , wetting of the solid
Common interactions that can occur[3] include: is said to take place. If y > 90 , wetting does not take
place. The term ‘‘wetting’’ refers to the displacement
1. An ion-exchange process from a surface of one fluid by another. It is most com-
2. An ion-pairing interaction monly applied to the displacement of air from a liquid
3. Acid–base interaction via either hydrogen bond- or solid surface by water or an aqueous solution. The
ing between substrate and adsorbate or Lewis term ‘‘wetting agent’’ is applied to any substance that
acid–Lewis base reaction increases the ability of water or an aqueous solution to
displace air from a liquid or solid surface.
For good wetting, cos y should be as close as possi-
ble to 1; that is, y should be as close as possible to 0.

A
60 γLA
γ (mNm–1)

γSA γSL

B
30 θ γLA
CMC

Log c γSL
γSA

Fig. 1 Schematic plot of surface or interfacial tension (g) vs. Fig. 2 Contact angles. In (A), y < 90 , and wetting of the
logarithm of the surfactant concentration (c). solid occurs; in (B), y > 90 , and wetting does not take place.
Surfactants in Pharmaceutical Products and Systems 3585

From Young’s equation, it can be seen that if gLA or The adsorption of surfactants onto solid surfaces is
gSL was minimized, cos y would be maximized, and important with respect to their detergent properties,
wetting would be promoted. their use as wetting agents in solid pharmaceutical dos-
Contact angles of water on powders of pharmaceu- age forms, and as stabilizers for suspension formula-
tical importance are usually measured by preparing tions. The mode of action of surfactants in each of
disks of the powder by compression or melting. these systems is discussed further below.
However, compaction may change the surface, so mak-

Suspensions
ing the measured result of little relevance. Contact

Super–
angles on finely divided solids can also be determined Micellization
by packing the powder into a tube and measuring the
penetration of liquids into the packing. As mentioned previously, surfactant molecules
Three types of wetting phenomena have been have the ability to form micelles in aqueous solution.
described:[4] adhesional wetting, spreading wetting, These micelles are colloidal-sized clusters of molecules.
and immersional wetting. Micellization is an alternative to interfacial adsorption
The way in which a particular system behaves for removing hydrophobic groups from contact
depends on the interfacial energies between the solid with the aqueous environment, thereby reducing
substrate and any contacting liquid, and between the the free energy of the system. In micelles, the hydro-
liquid and the second fluid (air). By manipulating these phobic groups are directed toward the center of the
factors, the wetting process can be controlled. This surfactant aggregate. In cases where there is little
may be achieved by the use of surfactants. distortion of the surrounding solvent by the hydro-
phobic group, there is little tendency for micellization
Modification of the wetting process by to occur, such as in water when the hydrophobic group
the use of surfactants of the surfactant is short or in the case of non-aqueous
solvents.
The effect of surfactants on the wetting process is a One of the most important applications of micelliza-
result of their adsorption at various interfaces with a tion in the context of pharmaceuticals is their ability to
resulting alteration of interfacial tensions. As has been solubilize drugs of poor aqueous solubility.
noted from Young’s equation, the wetting process is Micelles are dynamic species; there is a constant
promoted if either gLA or gSL or both are reduced with rapid interchange of surfactant molecules between
gSA remaining unchanged. Surfactants almost always the micelle and the bulk solution. Micelles cannot,
cause a reduction in gLA, however, the same cannot therefore, be regarded as rigid structures with a defined
be said for gSL and the effect on the interfacial tension shape, although an average micellar shape may be
depends on the nature of the adsorption. Thus the considered.
addition of a surface-active agent to the system does The main types of micelles recognized[3] are:
not always promote wetting, and spreading may in fact
be made more difficult.[4] 1. Small spherical
If adsorption of the surfactant molecules at the 2. Elongated cylindrical, rodlike micelles with
solid–liquid interface occurs in such a manner that they hemispherical ends (prolate ellipsoids)
are oriented with their polar ends toward the substrate 3. Large, flat lamellar micelles (disklike extended
and hydrophobic ends toward the liquid, the wettabil- oblate spheroids)
ity of an aqueous solution is reduced. This orientation 4. Vesicles, more or less spherical structures, con-
of surfactants molecules at the surface occurs if they are sisting of lamellar micelles arranged in one or
adsorbing to ionic or polar substrates (ion-exchange more concentric spheres.
or ion-pairing mechanism). However, at higher con-
centrations of surfactant, the surfactant ions adsorb In non-aqueous solvents, surfactants may form
by hydrophobic interaction with the already adsorbed ‘‘inverted micelles’’ where the hydrophilic heads of
layer, thus exposing their hydrophilic ends to the the surfactant molecules are present in the center of
solution in such a way that the surface becomes the micelle with the hydrocarbon chains extending out-
more readily wetted. Thus, the contact angle may first ward into the solvent. Dipole–dipole interactions hold
increase and subsequently decrease following the the hydrophilic heads of the surfactant molecules
addition of more surfactant to a solution. In contrast, together in the core, and in certain cases hydrogen
where adsorption occurs onto non-polar surfaces by, bonding between head groups can also occur.
for example, van der Waals attraction, the surfactant Micellar shape can be affected by changes in tem-
molecules are oriented with their hydrophilic groups perature, concentration and the presence of added
toward the liquid, the hydrophilicity of the substrate electrolyte to the liquid phase. Changes in any of these
is increased, and it becomes more wettable. factors may affect micellar size, shape, and aggregation
 3586 Surfactants in Pharmaceutical Products and Systems

number (number of surfactant monomers in the to ‘‘solubilize’’ both lipid soluble and water soluble
micelle). agents.
Several of the phase structures produced by surfac-
tants have potential as carriers and vehicles for drugs
Phase Behavior of Surfactants and also as targeting systems, used to direct the drug
to a specific site in the body.[5]
Equilibrium phase structures
Suspensions
Super–

As the concentration of a surfactant solution is SURFACTANT CLASSIFICATION

increased, structures of the types depicted in Fig. 3
may be encountered.[5] At concentrations well above Surfactant molecules may be classified based on the
the CMC, a more ordered structuring of the solution nature of the hydrophilic group within the molecule.
occurs. Two main types of liquid crystalline phases The four main groups of surfactants are defined as
may be identified: the middle phase, M, exhibiting follows:
a hexagonal array of indefinitely long, mutually paral-
lel rods; and the neat phase, G, with a lamellar struc- 1. Anionic surfactants, where the hydrophilic
ture. The liquid crystalline hexagonal phase, like the group carries a negative charge, such as car-
micellar phase, can exist either in a normal or reverse boxyl (RCOO
), sulphonate (RSO3
) or sul-
orientation. The order of phase structures formed phate (ROSO3
).
upon increasing surfactant concentration generally Examples of pharmaceutical importance include
follows a well defined sequence (Fig. 4) with a ‘‘mirror potassium laurate, CH3(CH2)10COO
Kþ, and
plane’’ through the lamellar phase in such a way sodium lauryl sulphate, CH3(CH2)11SO4
Naþ.
that normal phase structures can be considered to 2. Cationic surfactants, where the hydrophilic
be ‘‘oil-in-water’’ and the reverse structures to be group carries a positive charge (e.g., quaternary
‘‘water-in-oil.’’[5] ammonium halides, R4NþCl
). Examples of
pharmaceutical importance include cetrimide,
Modified phase structures a mixture consisting mainly of tetradecyl
(ca. 68%), dodecyl (ca. 22%), and hexadecyltri-
In addition to the equilibrium phase structures men- methylammonium bromides (ca. 7%), as well
tioned above, non-equilibrium surfactant phase struc- as benzalkonium chloride, a mixture of alkyl-
tures exist thatare also finding applications in drug benzyldimethylammonium chlorides of the gen-
delivery. Vesicular forms of surfactants are generally eral formula [C6H5CH2Nþ(CH3)2R]Cl
, where
formed by dispersing lamellar phases in an excess of R represents a mixture of the alkyls from
water (or non-aqueous polar solvents such as ethylene C8H17 to C18H37.
glycol or dimethylformamide) or, in the case of 3. Ampholytic surfactants (also called zwitterio-
reversed vesicles, in an excess of oil. With most surfac- nic surfactants), where the molecule contains,
tants, vesicles are non-equilibrium structures that will or can potentially contain, both a negative and
eventually re-equilibrate back into the lamellar phases a positive charge, (e.g., the sulfobetaines,
from which they originated. Vesicles are structural RNþ(CH3)2CH2CH2SO3
). Examples of phar-
analogs of liposomes (discussed later); they are maceutical importance include N-Dodecyl-N,
approximately spherical structures and have the ability N-Dimethylbetaine, C12H25Nþ(CH3)2CH2COO
.

Surfactant Spherical Rod-shaped Hexagonal Lamellar Reverse Reverse

Molecules Micelles Micelles Phase Phase Hexagonal Micelles
Phase

Fig. 3 Equilibrium phase structures of surfactant molecules. (From Ref.[5], reproduced by permission of the Royal Society of
Chemistry.)
Surfactants in Pharmaceutical Products and Systems 3587

Increasing surfactant concentration

'oil-in-water' 'mirror plane' 'water-in-oil'

H2O Micelle (L1) < Hexagonal (H1) < Lamellar (L0) < Reversed Hexagonal (H2) < Reversed Micelle (L2) Solid

Cubic (I1) Cubic (V1) Cubic (V2) Cubic (I2)

Suspensions
Fig. 4 Idealized phase sequence in surfactant-water systems. (From Ref.[5], reproduced by permission of the Royal Society of

Super–
Chemistry.)

4. Nonionic surfactants, where the hydrophile car- LIQUID SYSTEMS

ries no charge but derives its water solubility
from highly polar groups such as hydroxyl Solutions
or polyoxyethylene (OCH2CH2O–) groups.
Examples of pharmaceutical importance include Surfactants as solubilizing agents
polyoxyethylated glycol monoethers (e.g., cetoma-
crogol), sorbitan esters (SpansÕ) and polysorbates Solubilization can be defined as ‘‘the preparation of
(TweensÕ). a thermodynamically stable isotropic solution of a
substance normally insoluble or very slightly soluble
Tables 1–4 in the article Surfactants in Pharmaceu- in a given solvent by the introduction of an additional
tical Products and Systems in Volume 14 of the first amphiphilic component or components.’’[4] The
edition of this encyclopedia, together with the refer- amphiphilic components (surfactants) must be intro-
ences cited therein, give listings of some of the surfac- duced at a concentration at or above their critical
tants most commonly used in pharmaceuticals, along micelle concentrations. Simple micellar systems (and
with the purpose(s) for which they are usually reverse micellar) as well as liquid crystalline phases
employed. and vesicles referred to above are all capable of solubi-
lization. In liquid crystalline phases and vesicles, a ter-
nary system is formed on incorporation of the
SURFACTANT USES IN PHARMACEUTICAL solubilizate and thus these anisotropic systems are
PREPARATIONS not strictly in accordance with the definition given
above.[4]
Because of their unique functional properties, surfac-
tants find a wide range of uses in pharmaceutical pre- Solubilization by micelles
parations. These include, depending on the type of
product, improving the solubility or stability of a drug The location of a solubilized molecule in a micelle is
in a liquid preparation, stabilizing and modifying the determined primarily by the chemical structure of the
texture of a semisolid preparation, or altering the flow solubilizate. Solubilization can occur at a number of
properties of a granulate, thus aiding in the processing different sites in a micelle:
of the final tablet dosage form. In addition to their use
as excipients to improve the physical and chemical 1. On the surface, at the micelle–solvent interface,
characteristics of the formulation, surfactants may be 2. Between the hydrophilic head groups,
included to improve the efficacy or bioperformance 3. In the palisades layer, i.e., between the hydro-
of the product. The properties of surfactants are such philic groups and the first few carbon atoms
that they can alter the thermodynamic activity, solu- of thehydrophobic groups that comprise the
bility, diffusion, disintegration, and dissolution rate outer regions of the micelle core,
of a drug. Each of these parameters influence the 4. More deeply in the palisades layer, and
rate and extent of drug absorption. Furthermore, sur- 5. In the micelle inner core.
factants can exert direct effects on biological mem-
branes thus altering drug transport across the In aqueous systems, non-polar additives such as
membrane. The overall effect of inclusion of a surfac- hydrocarbons tend to be intimately associated with
tant in a pharmaceutical formulation is complex and the hydrocarbon core of the micelle. Polar and semi-
may be beyond those initially intended. Surfactants polar materials, such as fatty acids and alcohols are
may reduce the effectiveness of antimicrobials or pre- usually located in the palisades layer, the depth of pen-
servatives included in a formulation.[6] They also have etration depending on the ratio of polar to non-polar
the capacity to damage biological membranes. structures in the solubilizate molecule.
 3588 Surfactants in Pharmaceutical Products and Systems

In reverse micelles (formed in non-polar solvent capacity of the system, thereby greatly enhancing drug
systems containing surfactant), polar additives may solubility.
be solubilized in the core where a polar interaction of Solubilization has been used for many years in the
head groups occurs. formulation of phenolic antiseptic and disinfectant
A preferred location of the solubilizate molecule solutions. In the case of Cresol and Soap Solution
within the micelle is largely dictated by chemical struc- (Lysol) and Chloroxylenol Solution B.P., soap micelles
ture. However, solubilized systems are dynamic and are used to solubilize the phenolic substances. The
Suspensions

the location of molecules within the micelle changes soap (anionic surfactant) is formed by reaction of pot-
Super–

rapidly with time. Solubilization in surfactant aqueous assium hydroxide with a suitable oil such as linseed oil
systems above the critical micelle concentration offers (in Cresol and Soap Solution) or castor oil (in Chlor-
one pathway for the formulation of poorly soluble oxylenol Solution). The solubilizing potential of sur-
drugs.[6] From a quantitative point of view, the solubi- factant solutions for hydrophobic species has also
lization process above the CMC may be considered to been exploited in the design of cholelitholytic solvents
involve a simple partition phenomenon between an for gallstone dissolution with some limited success.
aqueous and a micellar phase. Thus the relationship
between surfactant concentration Cm and drug solubility Stability of drugs in solubilized systems
Ctot is given by Eq. (3).
Solubilization of a drug by incorporation into micelles
may affect its stability.[6] In the micelle, the molecular
Ctot ¼ Cs þ PCs Cm ð3Þ
environment of the drug molecules changes their prox-
imity and orientation with respect to each other, which
where Cs is the drug solubility in the absence of surface- may affect activity. In a micelle, the drug molecules
active agent and P is the distribution coefficient of drug may be protected from attacking species such as
between the micelle and bulk phases. A plot of Ctot versus hydronium or hydroxide ions and the stability of the
Cm is linear with a slope of PCs, which is the solubilizing drug may be increased. The difference in environment
capacity of the micelle.[7] between the micellar and bulk aqueous phases may be
The effect of altering the pH of the vehicle, in the such that reaction rates may be radically changed by
case of a partly ionized drug will be to alter the appar- the transfer of solute to micelles. Micellar systems
ent partition coefficient. Thus the effect of increasing may be used to deliberately alter the rates and direc-
the pH of a vehicle containing an acidic drug is to tions of chemical reactions.[6]
reduce the proportion of drug in the micellar phase.
If the surfactant is a weak electrolyte, it may induce AB block copolymer micelles
a concentration-dependent change in pH thus altering
drug partitioning and solubility.[8] It is well known that block copolymers in a selective
In general the solubilizing capacity for surfactants solvent (a good solvent for one block but a non-solvent
with the same hydrocarbon chain length increases in for the other) form a micellar structure through the
the order anionic < cationic < non-ionic, the effect association of the insoluble segments.[9] In contrast
being attributed to a corresponding increase in the area with micelles formed from low molecular weight sur-
per head group, leading to looser micelles with less factants, block copolymer micelles dissociate slowly
dense hydrocarbon cores which can accommodate to free polymeric chains. They have a greater capacity
more solute. for solubilizing aromatic molecules and express lower
The solubilizing capacity for a given surfactant CMCs. The AB block copolymers are considered use-
system is a complex function of the physicochemical ful vehicles for hydrophobic drugs.
properties of the two components which, in turn, influ- Only a few block copolymers form micellar struc-
ence the location or sites where the drug is bound to tures in aqueous milieu. One example is a series of
the micelle. The molar volume of the solubilizate polyethylene oxide/polypropylene oxide/polyethylene
together with its lipophilicity are important factors, oxide block copolymers known as Pluronics (trade-
the former reducing and the latter increasing solubili- name) or poloxamers. The poloxamers have been used
zation.[8] widely in pharmaceuticals, particularly as emulsifiers
Many pharmaceutical products contain a number of for intravenous lipids.[6] At low concentrations, polox-
solutes potentially capable of being solubilized within amer monomers are thought to form monomolecular
the micellar phase. Thus competition can occur micelles by a change in configuration in solution.[6]
between solutes resulting in an altered solubilizing At higher concentrations, aggregation of the monomo-
capacity. Furthermore, the addition of a second highly lecular micelles occurs. The aggregates so formed show
solubilized component to form a mixed micellar system the ability to solubilize drugs and increase the stability
may greatly alter the structure, size and solubilizing of solubilized materials. Poloxamers have low toxicity
Surfactants in Pharmaceutical Products and Systems 3589

and their solubilization capabilities might prove useful of surfactant ions to oppositely charged sites on the
in the delivery of hydrophobic drugs, although multi- particle surface, resulting in a lowering of the electrical
molecular micelle formation with core-shell structure energy barrier to the close approach of two particles to
is uncertain under physiological conditions.[10] each other. Flocculation may also occur by a bridging
Other block copolymers have been prepared and mechanism. A long (usually polymeric) surfactant mol-
studied as formulation adjuvants for hydrophobic ecule containing functional groups at various sites may
drugs, e.g., poly(ethylene oxide)/poly(aspartic acid) adsorb onto sites on the surface of adjacent particles,

Suspensions
and poly (ethylene oxide)/poly(b-benzyl-L-asparate) holding the particles together in a loose arrangement.

Super–
block copolymers have been used with adriamycin.[11,12] Alternatively if the surfactant molecules adsorb in such
a manner that the molecule extends into the liquid
phase, interaction of the extended portions of surfac-
Suspensions tant molecules adsorbed to different particles result
in bridging of those particles.
If a suspension is to be produced by a dispersion Another method of employing surfactants to
technique (as opposed to precipitation techniques), achieve flocculation is to first treat the particles with
surfactants may be used in the formulation to aid an ionic surfactant to disperse them. A readily soluble
dispersion of the solid particles in the liquid. This is electrolyte is then added which has the effect of com-
particularly important if the powder is not readily pressing the electrical double layer surrounding each
wetted by the liquid vehicle. Surfactants can reduce particle, allowing flocculation to occur. Subsequent
the interfacial tension between the solid particles and dilution of this type of system will redisperse it (due
the liquid vehicle. The advancing contact angle is to a decrease in electrolyte concentration).
reduced, and wetting of the solid particles promoted.
Such a system is said to be deflocculated. The inclusion
of a surface-active agent to improve powder wettability Emulsions
can often improve the bioavailability of the formu-
lation. Emulsification is one of the most important applica-
The forces at the surface of a particle affect the tions of surface-active agents in pharmaceutical
degree of flocculation and agglomeration in a suspen- systems. The phenomenon has been extensively studied
sion. Particles dispersed in a liquid medium may and many books and chapters of books have been
become charged in one of two main ways. Ionic species devoted to the subject.
present in solution may be adsorbed at the surface or, Macroemulsions are either oil in water (o/w) or
alternatively charges on the surface may arise due to water in oil (w/o). The type of emulsion formed
ionization of groups (such as carboxyl groups for depends largely on the emulsifying agent used; the pro-
example) which may be located at the surface. The cess and relative proportions of the oil and water
surface charge will influence the distribution of ions phases are less important. In general, o/w emulsions
in the aqueous medium surrounding the solid particles. are produced by emulsifying agents that are more sol-
The result is the formation of what is known as an uble in the water phase than in the oil phase, and w/o
‘‘electric double layer.’’ If the surface charge is posi- emulsions are produced by emulsifying agents that are
tive, immediately adjacent to the surface will be a more soluble in the oil phase. It is also possible to form
region of tightly bound solvent molecules and negative a multiple emulsion. For example, a small water or
counterions. Thus, the first layer is tightly bound, while aqueous solution droplet may be enclosed in a larger
the second layer (which still contains an excess of nega- oil droplet which is itself dispersed in an aqueous
tive ions) is more diffuse.[13] As two particles approach phase. Such a system is referred to as a ‘‘water-in-oil-
each other in aqueous medium, a weak attractive in-water’’ (w/o/w) emulsion. It is also possible to from
force exists just beyond the range of the double- an o/w/o emulsion.
layer-repulsive forces. This region is responsible for Many medicinal agents which have an unpalatable
the particle interaction termed ‘‘flocculation.’’ taste or texture can be made more acceptable for oral
Flocculated particles are weakly bonded, settle rap- administration when formulated as emulsions. Mineral-
idly, do not form a cake and so are easily resuspended. oil-based laxatives, oil soluble vitamins and high-fat
For this reason it is frequently desirable to promote nutritive preparations are frequently administered in
flocculation in a suspension. the form of o/w emulsions. It has been shown that in
The inclusion of surfactants in the formulation is some cases the absorption of drugs may be enhanced if
one way of achieving what is known as ‘‘controlled formulated as emulsions.[14] Emulsions (o/w) have also
flocculation.’’ Surfactants can cause dispersed solids been used for the intravenous administration of lipid
to flocculate by a number of different mechanisms.[3] nutrients. Radiopaque emulsions have been used as
The first is where there is an electrostatic attraction diagnostic agents in X-ray examinations.
 3590 Surfactants in Pharmaceutical Products and Systems

Emulsification is widely used in pharmaceutical pro- may arise either from adsorption of ions from the
ducts for external application such as lotions and aqueous phase or from frictional contact between drop-
creams, and in aerosol products to form foams. Semi- lets and the aqueous phase. In the latter case, the phase
solid emulsified formulations are discussed below. with the higher dielectric constant is positively
Based on the size of the dispersed particles or dro- charged.[3]
plets, emulsions may be classified[15] into
Microemulsions
Suspensions

1. Macroemulsions, droplets 0.1–50 mm, opaque

Super–

emulsions Microemulsions consist of large or ‘‘swollen’’ micelles,

2. Microemulsions, droplets 10–100 nm, trans- containing an internal phase similar to that found in a
parent dispersions. solubilized solution.[15] Unlike macroemulsions, they
appear as clear, transparent solutions. They tend to
Stabilization of the dispersion of one immiscible be more thermodynamically stable than macroemul-
liquid in another requires the addition of an emulsify- sions and can have essentially infinite lifetimes
ing agent which is commonly a surfactant or a mixture assuming no change in composition, temperature and
of surfactants. pressure. This is in contrast to macroemulsions which,
In the formation of an emulsion, one of the two although they may remain stable for long periods of
immiscible liquids is broken up into droplets which time, will ultimately undergo phase separation to
are dispersed in the other liquid. The dispersion of attain a minimum in free energy. Microemulsions can
one liquid in another immiscible liquid leads to a large generally be obtained by gentle mixing of the ingredi-
increase in interfacial free energy because of the ents of the emulsion. In this respect, they differ from
increase in the area of the interface. The emulsifying macroemulsions which require intense agitation for
agent stabilises the emulsion by adsorbing at the their formation. Microemulsions are usually prepared
liquid–liquid interface as an oriented interfacial film. with more than one surfactant or using a mixture of
This film reduces the interfacial tension between the surfactant and cosurfactant (e.g., a polar compound
liquids and also decreases the rate of coalescence of of intermediate chain length).
the dispersed droplets by forming mechanical, steric Microemulsions have been studied as drug delivery
and/or electrical barriers around them. systems, in particular for topical and transdermal drug
A strong mechanical barrier lessens the chance of delivery.[16,17]
droplets coalescing on collision. For maximum mech-
anical stability, the interfacial film of the adsorbed sur- Microspherical particles prepared by
factants should be close packed with strong lateral emulsification processes
interactions. For this reason, a mixture of two or more
surfactants is commonly used as the emulsifying agent, Emulsification–evaporation processes are widely used
such as a combination of a water-soluble surfactant in the preparation of polymer based microspherical
and an oil-soluble surfactant. In pharmaceutical (o/w) drug-loaded particulates. For example, hydrophobic
systems a mixture of a sorbitol ester (SpanÕ) with a drug-loaded PLA (polylactic acid) or PGLA (polylac-
polyoxyethyleneated sorbitol ester (TweenÕ) is often tide-co-glycolide) biodegradable microspheres are
used. The water soluble Tween tends to a have a great- often prepared from emulsions containing a non-
er interaction with the aqueous phase, its hydrophilic aqueous dispersed phase of dichloromethane contain-
group extending further into the water than that of ing the drug and polymer in an aqueous continuous
the non-oxyethyleneated ester. This is believed to phase. For the preparation of hydrophilic drug loaded
facilitate interaction of the hydrophobic groups of microspheres a double-emulsion process may be neces-
the molecules, as they can approach each other more sary. The nature of the surfactants used to stabilize
closely in the interfacial film. the emulsion phases can greatly influence the size,
The interfacial film can also stabilize the emulsion size distribution, surface morphology, loading, drug
by producing repulsive electrical forces between release, and bioperformance of the final multiparticu-
approaching droplets. This repulsion is due to surface late product.
charge on the droplets. The surface charge effect is
believed to be important only in the case of o/w emul-
sions. The source of surface charge is the hydrophilic Aerosols
head of the surfactant molecules which is oriented
toward the aqueous continuous phase. In emulsions Surfactants are found in both solution and suspension
containing ionic surfactant molecules, the charge on formulations of metered dose inhalers (MDIs). The
the disperse phase droplets is due to the amphipathic most common surfactants found in pressurised aerosol
ion. In the case of non-ionic surfactants, the charge preparations include sorbitan trioleate (Span 85),
Surfactants in Pharmaceutical Products and Systems 3591

oleic acid, and lecithins at concentrations of 0.1–2.0% wetting, solubilizing, detergent and penetration-
(w/w). These agents are non-volatile liquids which enhancing properties.
dissolve in the propellant blend. Their function in the Water-in-oil emulsions traditionally contain surfac-
formulation is to provide lubrication for the metering tants of natural origin such as cholesterol, wool fat,
valves and, in the case of suspension formulations, to wool alcohols, lanolin, divalent salts of fatty acids
maintain the disperse nature of the drug. soaps, calcium oleate and/or synthetic agents of low
The three surfactants commonly used in chlorofluro- hydrophilic-lipophilic balance (HLB) (indicating high

Suspensions
carbon (CFC)-based MDI formulations are insoluble lipophilicity), such as Spans (fatty acid esters of sorbi-

Super–
in the CFC-replacement propellants, hydrofluoroalk- tan). An example of such a product is Oily Cream B.P.
ane (HFA) 134a and HFA 227. Possible formulation which consists of a 1 : 1 mixture of wool alcohols and
alternatives involve the use of an adjuvant such as water.
ethanol to aid dissolution of the surfactant or a novel Oil-in-water creams, for topical use, generally con-
surfactant. Several companies have investigated novel tain mixed emulsifiers/surfactants; one of which is a
materials among which are fluorosurfactants, poly- water soluble surfactant with a high HLB, the other
oxyethylenes and drugs coated with surfactant.[18] being an amphiphile, usually a long chain fatty alcohol
(e.g., of chain length C14 to C18) or acid (e.g., palmitic
Controlled flocculation in metered-dose or stearic). The water soluble surfactant may be anio-
aerosol suspensions nic (e.g., sodium lauryl sulphate), cationic (e.g., cetri-
mide), or non-ionic (e.g., cetomacrogol, Tweens).
Controlled flocculation is a widely used technique for These mixed-surfactant systems are used not only
stabilizing suspended systems. The aim is to alter par- for their ability to form complex condensed films at
ticle surface charge or to achieve particle separation via the liquid–liquid interface, enhancing the stability of
steric hindrance with the help of appropriate stabiliz- the emulsion, but also because of their ability to impart
ing excipients. However this is particularly difficult to ‘‘body’’ to the product, resulting in a semisolid pro-
achieve in non-polar systems such as suspensions in duct rather than a liquid. Mixed emulsifiers control
CFC (or HFA) propellants. Controlled flocculation the consistency of a cream by forming a viscoelastic
to optimise the stabilisation of MDIs has been recom- network throughout the continuous phase of the emul-
mended by Ranucci et al.[19] but disputed by Hickey, sion. The network results from the interaction of
Dalby, and Byron.[20] the mixed emulsifier with water, forming a liquid crys-
talline phase.

Liposomes Foams

Liposomes are single- or multilayered phospholipids Emulsification is used in aerosol products to produce
vesicles. They are roughly spherical in shape and con- foams which are generally formulated as o/w emul-
sist of lipid bilayers alternating with aqueous regions. sions. The liquified propellant forms the disperse phase
Liposomes have shown potential as drug delivery of the emulsion, and the medication is usually in the
systems. The exact location of a drug molecule in a aqueous continuous phase. On discharge from the
liposome depends on its physicochemical composition pressurised container, the propellant vaporizes to form
and the composition of the lipids. Water soluble drugs bubbles which remain trapped within the aqueous
may be included in the aqueous phase, and oil-soluble phase giving rise to a foam. These are referred to as
drugs may be added to the membrane-forming phos- ‘‘stable foam’’ products. Nonaqueous stable foams
pholipid. An extensive account of the pharmaceutical may also be formulated, where the water is replaced
use of liposomes is found in the article ‘‘Liposomes by various glycols such as polyethylene glycol. ‘‘Quick
as Pharmaceutical Dosage Forms,’’ by Y. Barenholz, breaking foams’’ result when the propellant is in the
and D.J.A., Crommelin, Volume 9 of the first edition external phase. The product is emitted as a foam and
of this encyclopedia.[21] collapses into a liquid.

Biological Effects on Percutaneous Absorption

SEMISOLID SYSTEMS
Surfactants—traditionally common constituents and
Surfactants are major constituents of pharmaceutical, stabilizers of topical vehicles, ranging from hydro-
cosmetic, and food semisolid formulations, many of phobic agents such as oleic acid to hydrophilic
which are emulsions, either oil in water (o/w) or water sodium lauryl sulphate—have been tested as penetra-
in oil (w/o). They are included for their stabilizing, tion enhancers to improve transdermal drug delivery.
 3592 Surfactants in Pharmaceutical Products and Systems

Ionic surfactants are thought to enhance transdermal ‘‘boundary lubricant,’’ that is, the polar regions of
absorption by disordering the lipid layer of the stratum the molecule adhere to the metal surface of the die wall
corneum and by denaturation of keratin. The use of (in tablet manufacture). Adsorption of magnesium
penetration enhancers in general, and surfactants in stearate to the powder or granule surfaces also pre-
particular, in transdermal therapeutic systems has been vents agglomeration of the feed material and aids flow.
reviewed by Walters.[22] Lubricants may be classified as water-soluble or
water-insoluble. The latter are generally more effective
Suspensions

than water-soluble lubricants and can be used at a

Super–

SOLID DOSAGE FORMS lower concentration.[26] Common water-insoluble

lubricants (which are surfactants) include magnesium
Surface-active agents have been widely shown to stearate, calcium stearate, sodium stearate, and stearic
enhance drug dissolution rates. This may be due to acid; water-soluble lubricants include sodium lauryl
wetting effects, resulting in increased surface area, sulphate and magnesium lauryl sulphate.
effects on solubility and effective diffusion coefficient Sodium lauryl sulphate is used in the production of
or a combination of effects. Consequently surfactants hard gelatin capsules where it is added to the gelatin
have been included in tablet and capsule formulations solution during the preparation stage. The stainless
to improve wetting and deaggregation of drug particles steel molds are lubricated prior to dipping into the
and thus increase the surface area of particles available gelatin solution and sodium lauryl sulphate is added
for dissolution. to reduce the surface tension of the mix and cause
This wetting effect is found to be operative at con- the mold pins to wet more uniformly.[27]
centrations below the CMC. The effect of surfactants
on the dissolution of solids is complex. In addition to
effects on the available surface area, surfactants in Solid Dispersion Systems
concentrations above the CMC can increase drug
solubility and hence the effective concentration gradi- The bioavailability of hydrophobic drugs can be
ent. However they also reduce the effective rate of drug increased by strategies designed to enhance the dissolu-
diffusion as a consequence of drug solubilization tion rate of the drug. This has been achieved in many
within micelles. Models to quantify the effect of surfac- cases by forming a solid dispersion of the drug in a
tant concentration on drug dissolution have been suitable carrier, often a hydrophilic polymer such as
developed.[23] For solids whose dissolution is under sig- polyethylene glycol (PEG) or polyvinylpyrrolidone
nificant surface control, surfactants may further influ- (PVP). The drug is dispersed in the carrier by copreci-
ence the dissolution process. In this regard the pitation from a suitable solution containing both
enhancing effect of surfactants on the dissolution rate drug and carrier, by melting both components
of cholesterol has been widely studied.[24] together, or by some other process involving a phase
change. By using relatively high concentrations of car-
rier and a rapid precipitating process, the drug may
form as an amorphous or molecularly dispersed high
Hard Gelatin Capsules and Tablets energy phase in the carrier. A number of workers have
used surfactants as the carrier material to achieve this
Wetting agents enhanced dissolution effect. Among the surfactants
employed are polyoxyethylene stearate, Renex 650,
Surfactants are used in capsule[25] and tablet formula- poloxamer 188, Texafor AIP deoxycholic acid, and
tions as wetting agents to aid dissolution. Tweens and Spans. Surfactants have also been added
to conventional drug–polymer solid dispersions to
Lubricants, anti-adherents, and glidants further improve drug release properties. Sjokvist,
Nyström, and Aldén[28] found that the incorporation
The primary function of tablet lubricants is to reduce of sodium dodecyl sulphate (1–2%) in griseofulvin
the friction arising at the interface of tablet and die (3–10%)-PEG solid dispersions eliminated any traces
walls during compression and ejection. Lubricants also of crystalline drug, griseofulvin being present as a solid
possess antiadherent (prevention of sticking to the solution. Other three-component solid dispersions
punch and, to a lesser extent, to the die wall) and gli- containing surfactants have also been reported
dant (improvement of flow characteristics of powders such as Tween 20-Griseofulvin-PEG and Tween 20-
or granulates) characteristics and are useful in the pro- Oxodipine-PEG.[29] Problems have been reported
cessing of hard gelatin capsules. however as to the physical stability of surfactant
Magnesium stearate is used extensively as a lubri- containing systems, dissolution rates decreasing over
cant in tablet manufacture. It is an example of a a 12-month period.[30]
Surfactants in Pharmaceutical Products and Systems 3593

Matrix Systems of gastric emptying and retarding the movement of

drug to the absorption site by increasing the viscosity
Drug release from non-disintegrating inert matrices, of the formulation. This is thought to be especially true
fabricated from hydrophobic carriers such as poly- of polyoxyethylene derivatives. Bile salts, which are
ethylene, is improved by the presence of surfactants physiological surfactants, have been shown to affect
in the dissolution medium. Drug release was shown the rate of gastric emptying. The presence of bile salts
to be a function of the pore size distribution of the in the stomach has also been shown to affect ionic

Suspensions
matrix and the permeation pressure of the release movement across the gastric mucosa, thus increasing

Super–
media defined by its surface tension and contact angle. the movement of hydrogen and chloride ions out of
Inclusion of dioctyl sodium succinate, which reduced the lumen.
the contact angle below 90 , greatly enhanced drug Surfactants may also affect the rate and extent of
release; increasing the concentration of polysorbate drug absorption by exerting an influence on the per-
in the range of 0.001–0.1% had the same effect.[31] Sur- meability of the biomembrane. Competitive binding
factants have also been included in matrix-type drug of the surfactant to the membrane protein is con-
delivery systems to aid penetration of the dissolution sidered to be partially responsible for enhanced drug
medium thus increasing the rate and extent of drug absorption in many cases. Alternatively, the enhance-
release. ment may be due to allosteric rearrangement of the
membrane protein which is triggered by the binding
of one or more permeating species.
Suppositories
Nakanishi, Masada, and Nadai[33] studied the effect
of a range of surfactants on the rectal absorption of
Several non-ionic surface-active materials have been
sulphaguanidine and found absorption to be increased.
developed as suppositories vehicles. Many of these
The increase was associated with histological changes
bases, known as water-dispersible bases, can be used
in rectal membrane, increasing the rectal permeability.
for the formulation of both water-soluble and oil-
The same authors found that surfactants such as
soluble drugs.[32] The surfactants most commonly used
sodium deoxycholate and sodium dodecyl sulphate
are thepolyoxyethylene sorbitan fatty acid esters
used together with the chelating agent EDTA could
(Tweens), the polyoxyethylene stearates, and the sorbi-
increase the rectal absorption of macromolecules such
tan fatty acidesters (Spans). These surfactants may be
as inulin, insulin, and albumin.
used alone, blended, or with other suppository base
The membrane effects of surfactants are explained
materials to yield a wide range of melting points and
by a combination of membrane-surfactant binding,
consistencies.
disruption of membranes through solubilization into
Surface-active agents are widely used in combi-
lipoproteins, proteins, and mixed micelles, protein–
nation with other suppository bases. The inclusion of
protein interactions, and selective solubilization of
these agents in the formulation may improve the
some membrane components by the surfactant. The
wetting and water-absorption properties of the sup-
structure of the surfactant may play a role in determin-
pository. In addition, emulsifying surfactants help to
ing the range and extent of the influence of a particular
keep insoluble substances suspended in a fatty base
surfactant on drug absorption. It appears that the
suppository.[32]
greatest effect is achieved by molecules having a
The inclusion of a surfactant in the suppository for-
C12–C16 hydrocarbon chain, polyoxyethylene chain
mulation may enhance the rectal absorption of drugs.
lengths between 10 and 20, and molecular areas
The effect has been attributed to the formation of
between 1.0 and 1.6 nm2.[4] These effects, in the case
mixed micelles. It has been suggested that the presence
of drugs of low aqueous solubility, are in addition to
of the micelle facilitates the incorporation of the lipid
the higher absorption rate, arising from an increase
component of the mixed micelle into the biological
in drug solubility.[34,35]
membrane. This lipid then enhances the fluidity and
Surfactants, at high concentrations, exhibit some
permeability of the membrane to the poorly absorbed
toxicity and have the ability in many cases to disrupt
drug. It appears that the colorectal mucous membrane
a membrane. Both ionic and non-ionic surfactants
is more sensitive to the effects of mixed micelles than
have been shown to assist the breakdown of the
the gastrointestinal membrane of the small intestine.
mucous layer covering the epithelium and at high con-
centrations are thought to interfere with the membrane
Surfactant Influence on Drug Absorption from itself, which may lead to disruption of membrane
the Gastrointestinal Tract metabolism, particularly with regard to enzyme sys-
tems associated with the membrane. Adverse reactions
In the context of oral dosage forms containing surfac- to drug formulation agents including surfactants have
tants, these agents may play a role in reducing the rate been reviewed by Weiner and Bernstein.[36]
 3594 Surfactants in Pharmaceutical Products and Systems

DIRECT ACTIONS OF SURFACTANTS interfaces of the lung and lower surface tension at end-
expiration thus preventing alveolar collapse. If the
Detergents amount or quality of endogenous surfactant is inade-
quate, inspiratory pressure and the work of breathing
Detergents are surfactants that are used for the must increase in order to re-expand the alveoli with
removal of foreign matter from a solid surface. The each breath and permit adequate gas exchange. As
process involves many of the actions specific to surfac- the infant grows tired, progressive respiratory failure
Suspensions

tant molecules. The surfactant requires good wetting occurs.

Super–

properties to ensure good contact with the solid sur- Phosphatidylcholine is the major component of
face. It must also have the ability to remove dirt into endogenous surfactant, constituting about 60% of total
the bulk liquid. This is achieved by a lowering the phospholipids, and dipalmitoylphosphatidylcholine
dirt–liquid and solid–liquid interfacial tensions, thus (DPPC) is the primary surface-tension lowering
reducing the work of adhesion between the dirt and phospholipid.
the solid and enabling the dirt to be readily detached. The surfactant replacement therapy treatment used
Once detached, adsorption of surfactant at the dirt may be either ‘‘natural’’ or ‘‘artificial.’’ Natural sur-
particle surface prevents deposition, allowing the dirt factants are derived from bovine or porcine animal
to be washed away. If the dirt is oily it may be emulsi- lungs or human amniotic fluid. Synthetic or artificial
fied or solubilized by the surfactant. surfactants are composed of DPPC and spreading
agents such as unsaturated phosphatidylglycerol or
tyloxapol and hexadecanol.[39]
Antimicrobial Activity

Significant antimicrobial effects have been associated

NATURALLY OCCURRING SURFACTANTS
with cationic surfactants, in particular the quaternary
compounds. The action mechanism of quaternary sur-
Of the naturally occurring surfactants, the bile salts
factants involves disruption of the cell membrane, pro-
and phospholipids are of particular importance.
tein denaturation, and enzyme inhibition. Quaternary
compounds are able to lyse cells at relatively low con-
centration, resulting in leakage of cell contents into the Phospholipids
surrounding medium. Quaternary ammonium and
some phosphonium surfactants are used as topical The phospholipids are widely found in biological mem-
disinfectants in commercial dermatological products, branes and can be used as emulsifiers especially for
in surgical hand scrubs, and in the irrigation of skin intravenous fat emulsions, and as a key component
wounds. The most commonly used quaternary com- of liposomes. The elucidation of factors governing
pounds employed for theirantimicrobial effects are the solubilization of drugs in phospholipid dispersions
cetylpyridinium chloride, benzalkonium chloride, can provide some clues as to the biological role of
benzethonium chloride and cetyltrimethylammonium interactions with lipid systems in vivo.[4] Phospholipids
bromide.[37] Other surfactants, containing more than have been discussed above and in reference[21] in the
one quaternary (or positively ionizable group) are among context of liposomes.
the most active substances known in terms of anti-
microbial activity. Included in this group are dequali-
Bile Salts
nium acetate and chlorhexidine gluconate which have
been used in throat lozenges and mouthwashes. The
Bile salts are carboxylic acids (C22–C28) with a cyclo-
lysis of cells can also occur in the presence of anionic
pentenophenanthrene nucleus containing a branched
surfactants, although these are in general weaker in their
chain of 3–9 carbon atoms ending in a carboxyl group.
antimicrobial activity. A wide range of anionics, in parti-
Structurally they form micelles which are different
cular sodium lauryl sulphate and its homologs, finds wide
from the conventional spherical micelles synonymous
application in mouthwashes.[37]
with amphiphiles having a distinct hydrocarbon chain.
The hydrophobic feature of the bile salts is associated
Respiratory Distress Syndrome (RDS) with one surface of the steroid nucleus, and conse-
quently intermolecular association is much more
In 1959, surfactant deficiency was identified as the restricted. Primary and secondary micelles have been
major pathogenic factor in respiratory distress syn- proposed, the former consisting of two to four mole-
drome in infants.[38] Pulmonary surfactant is a complex cules, the latter being composed of aggregates of the
mixture of phospholipids, neutral lipids, and specific primary micelles. The CMC is less distinct and is
proteins which spread as a monolayer at the air-liquid highly dependent on the structure of the specific bile
Surfactants in Pharmaceutical Products and Systems 3595

salt, in particular the number of hydroxy groups and has been achieved after immunization with iscom-
their orientation. based vaccines, against viruses like the Epstein–Barr
Many studies have been completed in order to virus[42] and the measles virus.[43]
assess the effect of bile salts on the bioavailability of
poorly soluble drugs. Bile salts for example, have been
shown to enhance the absorption of sulphaguanidine
and urogastrone. Bile salts may also play a role in SURFACE ACTIVITY OF DRUGS

Suspensions
enhancing the transport of a compound from the

Super–
lumen of the intestine to the systemic circulation. Such A large number of drug molecules exhibit surface
absorption involves overcoming the resistance of the activity, that is, they tend to accumulate at interfaces,
aqueous boundary layer and the membrane epithelium depress surface tension and associate to form aggre-
to the passage of the drug. gates in solution. Although the hydrophobic groups
Bile salts readily form mixed micelles with lipid-like of most drugs are aromatic, they still behave like typi-
molecules such as lecithins or fatty acids. These mixed cal surfactants (which possess flexible hydrophobic
micelles are structurally very different from the simple chains), inasmuch as these aromatic groups have a high
micelles and generally have a much greater solubilizing degree of flexibility. (Drugs that exhibit association
capacity for hydrophobic molecules, both biological characteristics typical of surface active agents and
and synthetic. The solubility of DDT, a non-polar, may reduce surface tension are reviewed in Ref.[4].)
water insoluble molecule, for example, in bile salt Most of the drugs form micelles at concentrations
micellar solution can be increased to a far greater that they do not attain in vivo. It is therefore their sur-
extent by the addition of unsaturated long chain fatty face activity, rather than their self-association tendency
acids, probably because of mixed micelle formation. which is more important biologically. Surface-active
drugs will tend to bind hydrophobically to proteins
Saponins and other macromolecules and to associate with other
amphipathic substances such as bile salts, phospholi-
Saponins are glycosides found in certain plants which pids, and receptors. As with other surface-active
are characterized by their property of producing a agents, surface-active drugs may interact directly with
frothing aqueous solution. The term ‘‘saponin’’ is biological membranes. The possible biological implica-
derived from the Latin ‘‘sapo’’ meaning soap. Plant tions of surface activity is discussed by Attwood and
materials containing saponins have been used for a Florence[4] in relation to the phenothiazine tranquilli-
long time in many parts of the world for their detergent sers and local anesthetics.
properties, for example, in Europe, the root of Sapo-
naria officinalis and in South America the bark of
Quillaja saponaria.[40]
The saponin structure is either of the steroidal REFERENCES
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terpenoid type. Triterpenoid saponins are found, for 1. Rupprecht, H.; Lee, G. Adsorption at solid surfaces. In
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Quillaja saponaria and other species of Quillaja and 2. Myers, D. Surfaces and interfaces: general concepts. In
is used as an emulsifying agent. Liquorice, the root Surfaces, Interfaces, and Colloids: Principles and Appli-
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of which also contains triterpenoid saponins, has long 3. Rosen, M.J. Surfactants and Interfacial Phenomena;
been used in pharmacy as a flavoring agent, demulcent, Wiley Interscience: New York, 1989.
and mild expectorant. 4. Attwood, D.; Florence, A.T. Surfactant Systems. Their
Chemistry, Pharmacy and Biology; Chapman and Hall:
London, New York, 1983.
Iscoms 5. Lawrence, M.J. Surfactant systems: their use in drug delivery.
Chem. Soc. Rev. 1994, 23 (6), 417–424.
6. Florence, A.T. Drug solubilization in surfactant systems,
Iscoms (Immune-stimulating complexes) are stable drugs and the pharmaceutical sciences. In Techniques
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40 to 100 nm. They are promising carriers for antigens 7. Ong, J.T.H.; Manoukian, E. Micellar solubilization of timo-
in subunit vaccines. Iscoms are considered to be multi- besone acetate in aqueous and aqueous propylene glycol
micellar structures, shaped and stabilized by hydro- solutions of nonionic surfactants. Pharm. Res. 1988,
5 (11), 704–708.
phobic interactions, electrostatic repulsion, steric 8. Fahelelbom, K.M.S.; Timoney, R.F.; Corrigan, O.I.
factors and possibly hydrogen bonds.[41] Protection Micellar solubilization of clofazimine analogs in aqueous
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solutions of ionic and nonionic surfactants. Pharm. Res. 27. Jones, B.E. Gelatin additives, substitutes and extenders. In
1993, 10 (4), 631–634. Hard Capsules: Development and Technology; Ridgway,
9. Riess, G.; Hurtrez, G.; Bahadur, P. Block copolymers. In K., Ed.; The Pharmaceutical Press: London, 1987; 54.
Encyclopedia of Polymer Science and Engineering, 2nd Ed.; 28. Sjokvist, E.; Nyström, C.; Aldén, M. Physicochemical
Mark, H.F., Bikales, N.M., Overberger, C.G., Menges, G., aspects of drug release, 13. The effect of sodium dodecyl-
Eds.; Wiley-Interscience: New York, 1985; Vol. 2, 324–434. sulfate additions on the structure and dissolution of a
10. Kataoka, K.; Kwon, G.S.; Yokoyama, M.; Okano, T.; drug in solid dispersions. Int. J. Pharm. 1991, 69 (1),
Sakurai, Y. Block-copolymer micelles as vehicles for drug 53–62.
delivery. J. Control. Release 1993, 24 (1–3), 119–132. 29. Veiga, M.D.; Escobar, C.; Bernad, M.J. Dissolution beha-
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11. Yokoyama, M.; Miyauchi, M.; Yamada, N.; Okano, T.; vior of drugs from binary and ternary systems. Int. J.
Sakurai, Y.; Kataoka, K.; Inoue, S. Polymer micelles as Pharm. 1993, 93 (1–3), 215–220.
Super–

novel drug carrier—adriamycin-conjugated poly(ethylene 30. Sjokvist-Saers, E.; Nyström, C.; Aldén, M. Physicochemical
glycol) poly(aspartic acid) block copolymer. J. Control. aspects of drug release, 16. The effect of storage on drug
Release 1990, 11 (1–3), 269–278. dissolution from solid dispersions and the influence of cool-
12. Kwon, G.S.; Naito, M.; Yokoyama, M.; Okano, T.; ing rate and incorporation of surfactant. Int. J. Pharm.
Sakurai, Y.; Kataoka, K. Physical entrapment of adriamy- 1993, 90 (2), 105–118.
cin in AB block copolymer micelles. Pharm. Res. 1995, 31. Singh, P.; Desai, S.J.; Simonelli, A.P.; Higuchi, W.I. Role of
12 (2), 192–195. wetting on the rate of drug release from inert matrices. J.
13. Martin, A.N.; Bustamante, P. Physical Pharmacy: Physical Pharm. Sci. 1968, 57 (2), 217–226.
Chemical Principles in the Pharmaceutical Sciences, 4th Ed.; 32. Coben, L.J.; Lieberman, H.A. Suppositories. In The Theory
Lea and Febiger: Philadelphia, 1993; 386–388, 541–542. and Practice of Industrial Pharmacy, 3rd Ed.; Lachman,
14. Carrigan, P.J.; Bates, T.R. Biopharmaceutics of drugs L., Lieberman, H.A., Kanig, J.L., Eds.; Lea & Febiger:
administered in lipid-containing dosage forms I: GI absorp- Philadelphia, 1986; 564–588.
tion of griseofulvin from an oil-in-water emulsion in the rat. 33. Nakanishi, K.; Masada, M.; Nadai, T. Effect of pharmaceu-
J. Pharm. Sci. 1973, 62, 1476–1479. tical adjuvants on the rectal permeability to drugs. IV.
15. Eccleston, G. Encyclopedia of Pharmaceutical Tech- Effect of pharmaceutical adjuvants on the rectal per-
nology, 1st Ed.; Swarbrick, J., Boylan, J.C., Eds.; Marcel meability to macromolecular compounds in the rat. Chem.
Dekker, Inc.: New York, 1992; Vol. 5, 137–188. Pharm. Bull. 1984, 32, 1628–1632.
16. Osborne, D.W.; Ward, A.J.I.; O’Neill, K.J. Microemulsions as 34. O’Reilly, J.R.; Corrigan, O.I.; O’Driscoll, C.M. The effect
topical drug delivery vehicles I. Characterization of a model of simple micellar systems on the solubility and intestinal-
system. Drug Dev. Ind. Pharm. 1988, 14 (9), 1203–1219. absorption of clofazimine (B663) in the anesthetized rat.
17. Linn, E.E.; Pohland, R.C.; Byrd, T.K. Microemulsions for Int. J. Pharm. 1994, 105 (2), 137–146.
intradermal delivery of cetyl alcohol and octyl dimethyl 35. O’Reilly, J.R.; Corrigan, O.I.; O’Driscoll, C.M. The effect
PABA. Drug Dev. Ind. Pharm. 1990, 16 (6), 899–920. of mixed micellar systems, bile-salt fatty-acids, on the solu-
18. Bowman, P.A.; Greenleaf, D. Non-CFC metered dose inha- bility and intestinal-absorption of clofazimine (B663) in the
lers: the patent landscape. Int. J. Pharm. 1999, 86 (1), 91–94. anesthetized rat. Int. J. Pharm. 1994, 109 (2), 147–154.
19. Ranucci, J.A.; Dixit, S.; Bray, R., Jr.; Goldman, D. Appli- 36. Weiner, M.; Bernstein, I.L. Adverse Reactions to Drug
cation of controlled flocculation to metered dose aerosols Formulation Agents; Marcel Dekker, Inc.: New York, 1989.
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20. Hickey, A.J.; Dalby, R.N.; Byron, P.R. Effects of surfac- Forms: Disperse Systems, 2nd Ed.; Lieberman, H.A.,
tants on aerosol powders in suspension, implications for Rieger, M.M., Banker, G.S., Eds.; Marcel Dekker, Inc.:
airborne particle size. Int. J. Pharm. 1988, 42, 267–270. New York, 1996; Vol. 1, 211–286.
21. Barenholz, Y.; Crommelin, D.J.A. Liposomes as pharma- 38. Avery, M.E.; Mead, J. Surface properties in relation to atel-
ceutical dosage forms. In Encyclopedia of Pharmaceutical ectasis and hyaline membrane disease. Am. J. Dis. Child.
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Marcel Dekker, Inc.: New York, 1994; Vol. 9, 1–39. 39. Fujiwara, T.; Maeta, H.; Chida, S.; Morita, T.; Watabe, Y.;
22. Walters, K.A. Penetration enhancers and their use in trans- Abe, T. Artificial surfactant therapy in hyaline-membrane
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Hadgraft, J., Guy, R.H., Eds.; Drugs and the Pharmaceuti- 40. Evans, W.C. Saponins, cardioactive drugs and other ster-
cal Sciences; Marcel Dekker, Inc.: New York, 1989; Vol. 35, oids. In Trease and Evans’ Pharmacognosy, 14th Ed.; WB
197–246. Saunders Company Ltd.: London, 1996; 293–309.
23. Higuchi, W.I. Diffusional models useful in biopharmaceu- 41. Kersten, G.F.A. Aspects of Iscoms. Stuctural, Pharmaceuti-
tics. Drug release rate processes. J. Pharm. Sci. 1967, 56, cal and Adjuvant Properties. Ph.D. thesis, University of
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24. Higuchi, W.I.; Su, C.C.; Park, J.Y.; Gulari, E. Mechanism 42. Morgan, A.J.; Finerty, S.; Lövgren, K.; Scullion, F.T.;
of cholesterol gallstone dissolution. Analysis of the kinetics Morein, B. Prevention of epstein-barr (EB)virus-induced
of cholesterol monohydrate dissolution in taurocholate/ lymphoma in cottontop tamarins by vaccination with the
lecithin solutions by the mazer, benedek, and carey models. EB virus envelope glycoprotein GP 340 incorporated into
J. Phys. Chem. 1981, 85 (2), 127–129. immune-stimulating complexes. J. Gen. Virol. 1988, 69,
25. Newton, J.M.; Rowley, G.; Törnblom, J.-F.V. Further stud- 2093–2096.
ies on the effect of additives on the release of drug from 43. de Vries, P.; van Binnendijk, R.; van der Marel, P.; van
hard gelatin capsules. J. Pharm. Pharmac. 1971, 23 (suppl.), Wezel, A.L.; Voorma, H.O.; Sundquist, B.; UytdeHaag,
156S–160S. F.G.C.M.; Osterhaus, A.D.M.E. Measles virus fusion pro-
26. Banker, G.S.; Peck, G.E.; Baley, G. Tablet formulation tein presented in an immune-stimulating complex (iscom)
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Suspensions
Robert A. Nash
Consultant, Mahwah, New Jersey, U.S.A.

Suspensions
Super–
INTRODUCTION contain 125–500 mg of active solid material in a 5-ml
(teaspoonful) dose, whereas a drop concentrate may
A suspension is a particular class or type of dispersion provide the same amount of insoluble drug in a 1- or
or dispersed system in which the internal or suspended 2-mL dose. Antacids and radiopaque suspensions con-
phase is dispersed uniformly by mechanical agitation tain relatively high amounts of suspended material for
throughout the external phase (called the suspending oral administration. The vehicle may be a syrup, a sor-
medium or vehicle). The internal phase, consisting of bitol solution, or a gum-thickened, water-containing
a homogeneous or heterogeneous distribution of solid artificial sweetener because in addition to ingredients,
particles having a specific size range, is maintained uni- safety, taste, and mouthfeel are important formulation
formly throughout the suspending vehicle with the aid considerations. In the case of limited shelf life (low
of a single or a particular combination of suspending chemical stability of the insoluble drug), the dosage
agent(s). In addition, unlike in a solution, the sus- form may be prepared as a dry granulation or powder
pended particles exhibit a minimum degree of solu- mixture that is reconstituted with water prior to use.
bility in the external phase. In a colloidal suspension,
the solids are less than about 1 mm in size. In a coarse
suspension, they are larger than about 1 mm. The prac- Topical Suspensions
tical upper limit for individual suspendable solid parti-
cles in coarse suspensions suspensions is approximately Historically, the externally applied ‘‘shake lotion’’ is
50–75 mm. When one or more types of solid particles the oldest example of a pharmaceutical suspension.
that constitute the internal phase are pharmaceutically Calamine Lotion USP, as well as other dermatological
useful and/or physiologically active, the system is preparations, is closely associated with the technical
known as a pharmaceutical suspension. development of the pharmaceutical suspension.[2]
In an emulsion, the particles of the internal phase Because safety and toxicity considerations are most
are spherical or liquid droplets that are dispersed readily dealt with in terms of dermatological accept-
throughout a liquid external phase. Even though the ability, many useful suspending agents were first intro-
particles may be liquid only at elevated temperatures duced in topical formulations.[3] In addition, the
(50–80
C) and semisolid or rigid at room temperature, protective action and cosmetic properties of topical
as long as they appear spherical on careful microscopic lotions usually require the use of high concentrations of
examination, they are generally considered to be emul- the dispersed phase, often in excess of 20%. Therefore,
sified rather than suspended. Thus, a clue to the pres- topical lotions represent the best example of suspensions
ence of a suspended particle is its lack of sphericity that exhibit low settling rates.[4] Various pharmaceutical
or its definitive lattice structure. Exceptions to this gen- vehicles have been used in the preparation of topical
eral rule are spherical microspheres and related spheri- lotions, including diluted oil-in-water or water-in-oil
cal solid microparticles. emulsion bases, determatological pastes, magmas, and
clay suspensions.

CLASSIFICATION
Parenteral Suspensions
Martin and Bustamante[1] list three general classes
The solids content of parenteral suspensions is usually
of pharmaceutical suspensions: orally administered
between 0.5 and 5.0%, except for insoluble forms of
(sometimes referred to as mixtures); externally applied
penicillin, in which concentrations of the antibiotic
(topical lotions); and injectable (parenteral).
may exceed 30%. These sterile preparations are
designed for intramuscular, intradermal, intra-lesional,
Oral Suspensions intra-articular, or subcutaneous administration. The
viscosity of a parenteral suspension should be low
The solids content of an oral suspension may vary con- enough to facilitate injection. Common vehicles for
siderably. For example, antibiotic preparations may parenteral suspensions include preserved 0.9% saline
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-100200021
Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved. 3597
 3598 Suspensions

solution or a parenterally acceptable vegetable oil. The a large number of low-density organic materials and
primary factor governing the selection of injectable pharmaceutical substances, such as charcoal and sul-
ingredients is safety. Ophthalmic suspensions that are fur. The hydrophobic nature of the latter group is
instilled into the eye must be prepared in a sterile accentuated by entrapped air adsorbed on the surface
manner. The vehicle employed is essentially isotonic of these particles. Hydrophobic materials may be
and aqueous in composition. wetted by oils and semipolar liquids and are called
lipophilic solids; conversely, hydrophilic materials
Suspensions

behave like lipophobic solids in oils.

Super–

UTILITY OF SUSPENSIONS Hydrophilic solids can be suspended easily in water

without the aid of a water-dispersible surfactant or
A suspension is often chosen as pharmaceutical dosage wetting agent, and conversely hydrophobic solids can
form for drugs insoluble in water and aqueous fluids be suspended in oils and non-polar vehicles without the
at the dosage required for administration and when use of lipid-soluble surfactants. The crystal density of
attempts to solubilize the drug would compromise hydrophilic solids usually ranges from 1.5 to 6.9 g/cm3,
stability and safety. For oral administration, the taste whereas the crystal density of hydrophobic solids usually
of a bitter or unpleasant drug can often be masked ranges from 0.9 to 2.2 g/cm3.
by choosing an insoluble form of the active drug.
An aqueous suspension is a useful oral dosage form
for administering insoluble or poorly soluble drugs.
PARTICLE SIZE CONSIDERATIONS
The large surface area of the dispersed drug particles
often facilitate absorption. Unlike drug particles con-
The mean particle diameter and the particle size distri-
tained in tablets or capsules, the dissolution of drug
butions of suspended insoluble drugs are important
particles in suspension and subsequent absorption
considerations in formulating stable pharmaceutical
commence upon dilution in gastrointestinal fluids.
suspensions. Hiestand[5] defines the lower limit of
Finely divided particles dissolve faster and have higher
coarse suspensions as those containing particles larger
relative solubilities than do similar macroparticles.
than 0.1 mm. Except for a number of clays, oxides,
The parenteral suspension is an ideal dosage form
charcoal, and pigments, the average particle size of
for prolonged or ‘‘depot’’ release. In the administra-
most drugs and pharmaceutical rarely falls below
tion of a drug as an aqueous or oleaginous suspension
1 mm. Although most submicron inorganic excipients
into subcutaneous or muscular tissue, the drug is
appear to behave like hydrophilic solids, most insolu-
deposited at the injection site. The depot acts as a
ble drugs and pharmaceutical excipients are usually
reservoir, slowly releasing drug at a rate related to both
soft, organic, essentially crystalline hydrophobic solids
the intrinsic aqueous solubility of the drug form and
ranging in particle size from several microns to several
the type of suspending vehicle, either aqueous or oily
hundred or more.
for the purpose of maintaining prolonged systemic
Drug particle size is an important factor influencing
absorption of the drug from the injection site.
product appearance, settling rates, drug solubility,
in vivo absorption, resuspendability, and overall stabil-
ity of pharmaceutical suspensions. Insoluble drug par-
HYDROPHILIC/HYDROPHOBIC SOLIDS
ticles are seldom uniform spheres or cubes, even after
size reduction and classification. Wide distributions
Insoluble solids, regardless of particle size, that have a
in particle size often lead to high-density suspensions.
relatively low interfacial tension and are readily wetted
Systems with widely differing particle shapes (plates,
by water are called hydrophilic solids. These solids
needles, filaments, and prisms) frequently produce
include clays (bentonite, kaolin, talc, magnesium
low-density slurries. The growth over time of unprotec-
aluminum silicate); bismuth salts, barium sulfate, car-
ted, slightly soluble drug solids and changes in their
bonates, hydroxides, or oxides of calcium, magnesium,
particle size distribution in suspension are a serious
zinc, and aluminum; and titanium dioxide. The hydro-
problem. Crystal growth of particles is usually attribu-
philicity of a powder surface can be investigated with
ted to one or more of the following mechanisms:
the help of moisture absorption studies in which the
solid particles are exposed to varying relative humid-
ities. Insoluble powders that absorb moisture below  ‘‘Oswalt ripening’’ or the growth of large particles
relative humidities of 70–80% at room temperature at the expense of smaller ones, because of a dif-
are said to be hydrophilic solids. ference in solubility rates of different size particles.
Fine insoluble solids that are not easily wetted by For example, the increase in the solubility rate of
water and have a relatively high interfacial tension a 0.2-mm particle, is 13%. For a 2-mm particle, it is
are referred to as hydrophobic solids. These include 1%, and for particles above 20 mm, it is negligible.
Suspensions 3599

 Crystal growth due to temperature fluctuations on 11. Impurities and foreign substances during crys-
storage is of minor importance, unless suspensions tallization affect the reproducibility and aggre-
are subjected to temperature variations of 20
C or gation potential of many drug particle systems.
more. 12. Constant crystallizing conditions are essential.
 A polymorphic form may change to another more Batch-to-batch variation in crystal size and
stable crystalline form; changes in crystal habit shape is often associated with poor control of
may be related to the degree of solvation or processing and crystallization procedures.

Suspensions
hydration.

Super–
 Crystal growth may also arise when the more ener- Variations in assay results can be avoided by the
getic amorphous or glassy forms of a drug exhibit preparation of homogeneous, well-mixed, or non-settling
significantly higher initial solubility in water than fine particle suspensions (size 1–10 mm). Particle size
the corresponding crystalline forms. reduction results in slow, more uniform settling rates.
 Size reduction by crushing and grinding can produce The bioavailability of drugs is improved by reducing
particles whose different surfaces exhibit high and the size of suspension particles. Furthermore, drug
low solubility rates. This effect can be related to dif- particles smaller than 20 mm produce less pain and
ferences in the free surface energy introduced during tissue irritation when injected parenterally. However,
comminution (grinding). fine particles may have a deleterious effect on chemical
stability because of their high dissolution rate.
Crystal growth and changes in particle size distri-
bution can generally be controlled by employing one Particle Size Reduction
or several of the following procedures and techniques.
Drug solids are easy to grind. Reduction to a particle
1. Selection of particles with a narrow range of size of about 50–75 mm usually produces a free-flowing
particle sizes, such as microcrystals between 1 powder. Solids containing particles smaller than 50 mm
and 10 mm. tend to aggregate or agglomerate in the dry state. Fur-
2. Selection of a stable crystalline drug form that thermore, below 10–50 mm, the increased free surface
usually exhibits lower solubility in water. The energy, as evidenced by the cohesion of small particles,
crystalline form that is physically most stable becomes a factor interfering with further size reduction.
usually has the highest melting point. The powder may become damp, especially if there
3. High-energy milling should not be used during is a tendency to attract moisture. Material tends to
particle size reduction. Microcrystals are best ‘‘ball up,’’ which indicates that the agglomerated
formed by controlled precipitation techniques masses are larger than the individual particles con-
or shock cooling. tained within.
4. A water-dispersible surfactant wetting agent As the pores between powder particles become
dissipates the free surface energy of particles smaller with decreasing particle size, increases in
by reducing the interfacial tension between the surface area facilitate liquid penetration. Aggregates
solid and the suspending vehicle. behave like hydrophobic solids, entrapping air and
5. A protective colloid, such as gelatin, gum, or a becoming difficult to wet.
cellulosic derivative, is used to form a film bar- The most efficient method of producing fine parti-
rier around the particles, inhibiting dissolution cles is by dry milling prior to manufacture of the sus-
and subsequent crystal growth. pension. Dispersion equipment, such as colloid mills
6. The viscosity of the suspending vehicle is or homogenizers are normally used to wet-mill finished
increased to retard particle dissolution and sub- suspensions to break up poorly wetted fine particle
sequent crystal growth. aggregates or agglomerates. Among the several meth-
7. Temperature extremes during product storage ods of producing small, reasonably uniform drug
(freeze–thaw conditioning) must not occur. particles are micropulverization, fluid energy grinding,
8. Supersaturation favors the formation of needle- controlled precipitation, and spray drying. Fig. 1 illus-
like crystals and should be avoided. trates the four basic types of size reduction equipment
9. Rapid or shock cooling and high agitation favor used in the pharmaceutical industry to produce fine
the formation of thin, small crystals and should powder particles.
be avoided. Slow crystallization by evaporation
yields compact crystals. Micropulverization
10. Experimentation with different crystallizing sol-
vents is recommended to change crystal size Micropulverization is one of the most rapid, con-
and shape. venient, and inexpensive methods of producing fine
 3600 Suspensions

A B However, because it is important that a majority of

drug particles in parenteral suspensions are below
10 mm, fluid energy grinding is the most convenient
method for their production.

Controlled crystallization
Suspensions

A solvent that dissolves a solid readily at room tem-

Super–

perature may serve as a crystallizing medium when

mixed with another solvent in which the compound
is only sparingly soluble. A solution that is nearly satu-
rated at a temperature about 10
C below the boiling
point of the solvent combination is prepared in a tem-
C D perature ranging between 60 and 150
C. Separation of
microcrystals from such hot concentrated solutions is
commonly induced by cooling and stirring. However,
when supersaturation is obtained by agitation and
shock cooling of the hot solution and through the
rapid introduction of another cold miscible solvent in
which the drug is only sparingly soluble, formation
of minute crystalline particles (nucleates) proceeds
without appreciable crystal growth, and uniform
microcrystals of the drug are thus obtained. In
addition, ultrasonic methods have been used during
shock cooling to promote microcrystal formation.
Fig. 1 The four basic types of size reduction equipment used
to produce fine solid particles: (A) crushers and shredders; Spray drying
(B) hammermills; (C) colloid mills; and (D) fluid energy mills.
Particles of microcrystalline size can also be obtained
by spray-drying procedures, resulting in a porous,
drug powders. The milling equipment includes ham- free-flowing, easily wetted, essentially monodispersed
mermills, micropulverizers, universal mills, end-runner powder. With proper control of process variables,
mills, and ball mills. Micropulverizers are highspeed spherical particles are obtained that may be coated
attrition or impact mills specially adapted for fine with agents to aid suspension and promote stability.
grinding. Some mills are fitted with classifiers to facili- However, the process is not normally considered for
tate particle separation by centrifugation. Because the preparation of ultrafine powders.
ultrafine particles smaller than 10 mm are rarely pro-
duced, buildup of electrostatic charge on the surface
of milled powder is encountered only occasionally. PHYSICAL ASPECTS
The main disadvantage of micropulverization is the
large distribution of particle sizes produced, normally Stability of Suspensions
in the range of 10–50 mm or higher. Nevertheless, these
powders are satisfactory for the preparation of most The chemical stability of a drug in suspension is con-
oral and topical suspensions. trolled by the fact that the rate of degradation is
related to the concentration of the drug in aqueous
Fluid energy grinding solution rather than to the total concentration of the
drug in the product. Generally, a suspended drug
The process of fluid energy grinding, also referred to as decomposes only in solution as the solid phase gradually
jet milling or micronizing, is the most effective method dissolves; that is, a solution concentration equal to the
for reducing particles below 10 mm. The ultrafine parti- solubility of the drug is maintained. Drug degradation
cles are produced by the shearing action of high- in a suspension usually follows zero-order kinetics,
velocity streams of compressed air on particles in a with the rate constant solely dependent on the satu-
confined space. The main disadvantage of fluid energy ration solubility of the drug in solution. Reducing the
grinding is the high electrostatic charge built up on solubility of the suspended drug decreases the rate
the surfaces of the milled powder, which makes pow- of degradation. The constancy of potency may be
der classification and collection exceedingly difficult. improved by selecting a pH value or range where the
Suspensions 3601

drug is least soluble or by replacing the drug with a less are appreciable. Such conditions frequently lead to the
soluble derivative or salt. Decomposition in suspen- undersirable phenomenon of ‘‘caking or claying’’ and
sions may also be described as a diffusion-controlled require extensive agitation for resuspension.
process or by catalysis initiated by environmental The physical instability of these early deflocculated
factors such as oxygen, light, and trace metals. suspensions led to other methods of producing physi-
As a rule, the problem of suspension stability is cally stable pharmaceutical suspensions. For example,
complicated by the fact that pharmaceutical suspen- the density of the vehicle was made to equal or

Suspensions
sions are affected at least as much by physical as by approach the crystal density of the suspended drug

Super–
chemical factors. particles. If the drug particles are small enough and
Because a suspension exists in more than one state the vehicle sufficiently viscous, the particles remain
(liquid and solid), there are different ways in which suspended indefinitely in accordance with Stokes’ law.
the system can undergo chemical or physical change. Because the crystal density of most organic drug parti-
Some of the more obvious difficulties involved in cles lies somewhere between 1.1 and 1.5 g/cm3, the only
stability predictions are based on the fact that simple liquid vehicles for oral use with densities (at 25
C)
hydrostatic relationships (Stoke’s law, etc.) used to high enough to be considered are Sorbitol Solution
define settling ratesassume a spherical, deflocculated, USP (1.29 g/cm3), Syrup USP (1.31 g/cm3), and high-
free-falling particle that is not affected by particle– fructose corn syrup (1.41 g/cm3). In practice, however,
particle or particle–vehicle interactions. Suspensions it is extremely difficult to prepare oral suspensions by
that exhibit non-Newtonian floware difficult to define the matched-density technique alone because dilution
in terms of the basic stability relationships. In addition, with water and other liquids reduces the vehicle den-
suspensions that are described interms of a single sity. Nevertheless, the use of high-density liquids as
representative particle do not reflect theinfluence of suspending vehicles often has a beneficial effect on
the entire particle size distribution. physical stability.
Chemical stability predictions are sometimes com- The approximate settling velocities for non-floccu-
plicated by the difficulty of determining the pH value lated particles of various average size were determined
of suspensions, which often changes because of surface in the range of 0.2–200 mm at density differences
coating of electrodes and differences between bulk- between solid particles and the suspending liquid of 0.2
suspension and supernatant-vehicle readings. Acceler- and 2.0 g/cm3, respectively, with an absolute viscosity
ated elevated temperature stability testing often has a of the suspending liquid between 1 and 1000 centipoise
pronounced adverse effect on viscosity, particle solubil- (cP or mPa s). Terminal settling rates were calculated
ity, and size distribution. by a method described by Carpenter[6] for two concen-
trations of suspended solids, namely less than 2 and
20% vol. According to the analysis, permanent-type
Deflocculated Suspensions suspensions, which exhibit a settling viscosity of less
than 0.14 cm per year at 25
C, can be obtained with
The empirical method of producing pharmaceutical a suspending liquid of a viscosity of ca 1000 cP, a
suspensions is based on an attempt to prepare a stable density difference of 0.2 g/cm3 or less and an average
deflocculated dispersion of a drug in a suitable suspen- particle size of the suspended solid of 0.2 mm. This is a
sion vehicle. In the past, a series of suspensions was difficult set of criteria for most pharmaceutical suspen-
often prepared using different concentrations of a sions to meet.
favored suspending agent to identify the formulation Because the distribution of particle sizes in most
that would produce the most homogeneous looking pharmaceutical suspension is generally above 1 mm,
(‘‘smooth’’) and stable suspension. The finished prep- deflocculated or peptized systems settle very slowly in
aration was usually passed through a homogenizer or stages, with the larger particles settling more rapidly
colloid mill to improve the final dispersion. Smooth than smaller ones. Ultimately, they form a tight, dense
looking viscous suspensions were produced; however, sediment that is difficult to resuspend. When viewed
after some time, the drug particles settled slowly, form- under a microscope, the dispersed suspension consists
ing a tightly packed sediment that was almost impos- of individual particles, showing no apparent association.
sible to resuspend even with vigorous shaking. Primary Deflocculated suspensions are produced by three
particles or small aggregates, reaching the bottom of methods.
the container during sedimentation (settling), slipped
past each other and produced compact layers of solids. Mutual repulsion to large z-potential
The interparticle interaction in such compact sedi-
ments is relatively high because the interparticle dis- This is best achieved by the adsorption of an electro-
tances are small, and the weak van der Waals forces of lyte (KCI) or polyelectrolyte dispersant (sodium
attraction, which decreases exponentially with distance, hexametaphos-phate) on the surface of suspended
 3602 Suspensions

particles to create a strong mutual repulsion between Flocculated Suspensions

the microsize suspended particles. For example, mod-
erate stability is achieved when the z-potential is Matthews and Rhodes,[8] Haines and Martin,[7] Hie-
between 30 and 60 mV, and good to excellent physi- stand,[5] and Ecanow and co-workers[9,10] are credited-
cal stability is achieved when the z-potential is between with establishing the ‘‘structured particle’’ concept or
60 and 100 mV. As the size and density of the sus- flocculated pharmaceutical suspension. The following
pended particles increase beyond 1 mm and 1.0 g/cm3, definitions should prove useful in avoding confusion
Suspensions

respectively, the effect of z-potential becomes less among three closely related terms: flocculation, agglom-
Super–

important. eration, and coagulation. The term aggregation can

apply to all three.
Flocculation refers to the formation of a loose
Adsorption of a smaller hydrophilic or lyophilic
aggregation of discrete particles held together in a
colloid on larger suspended particles
networklike structure by physical adsorption of
macromolecules, bridging during chemical interaction
When a strongly hydrated hydrophilic protective
(precipitation), or when the longer-range van der
colloid, such as gelatin, is adsorbed on the surface of
Waals forces of attraction exceed the shorter-range
the suspended particles, the affinity for water exceeds
forces of repulsion. The floccule referred to as a ‘‘stable
the mutual attraction of adjacent particles for each
floc’’ usually contains varying amounts of entrapped
other. The protective colloid and hydrogen-bonded
liquid medium or vehicle within the networklike struc-
water molecules form a protective hydration layer
ture (Table 1).
around each suspended particle.
In agglomeration, a large number or mass of parti-
cles are closely bound together as aggregates in a dry
Steric hindrance due to adsorption of an oriented (air) or liquid state. Coagulation or severe overfloccu-
non-ionic surfactant or polyelectrolyte lation refers to the massing of particles in a liquid state
alone and sometimes in the form of a fluid gel struc-
Adsorption of a non-ionic polymer (gum or cellulosic) ture. Aggregated particles of overflocculated systems,
or surfactant (polysorbate 80) of sufficient chain length including adsorbed surface films, are in surface contact
creates steric hindrance and prevents adjacent sus- with each other, and each mass or coagula acts as
pended particles from coming close enough to join a unit. The particles of such coagulated systems are
each other. Steric stabilization has the advantage over held together by strong film-to-film bonds. Coagulated
electrostatic stabilization in that it is relatively insensi- suspensions, like deflocculated suspensions, tend to
tive to the presence of electrolyte in the aqueous ‘‘cake’’ on standing.
vehicle. Soon after milling and suspension, unless steps are
Because many pharmaceutical suspensions are not taken to prevent it, microsize particles tend to grow
capable of reaching a state of complete electrostatic with time. Because the solubility rate of unprotected
repulsion and producing and maintaining defloccu- particles (Ostwald ripening) is higher than that of
lated particles, the technique has been regarded as large crystals, crystal growth is favored until a more
unworkable by many investigators.[1,5] thermodynamically stable distribution of particles

Table 1 Suspension characteristics

Relative Sediment volume
settling in vials at Drainage
Type f-Potential rate equilibrium from vial Resuspendability
Agglomerated or coagulated 0 Non-uniform Low (lumpy) Poor Poor (may cake)
Overflocculated 0 Very high Very high Poor Good
Slightly over flocculated 0 High Moderate Fair Good
to high
Slightly underflocculated 0 Moderate Moderate Good Good
Underflocculated 0 to þ10 mV Slow Moderate Good Fair
to low
Deflocculated or peptized þ10 to þ30 mV Non-uniform Low (stratified) Good Poor
(cakes difficult to
resuspend)
Suspensions 3603

sizes is achieved. This phenomenon tends to promote Some authors[11] refer to the ‘‘stable floc’’ as the
‘‘caking and cementing together’’ of particles. partially flocculated state. Simply stated, the greater
The creation of a protective coat or boundary layer the number of particle-to-particle contact points in
(with a hydrophilic colloid) about such particles offers the cluster, the higher the degree of flocculation.
the best protection to crystal growth. Because protec- The main advantages of the stable floc are as
tive barriers may or may not flocculate, the substrate follows.
particles, the sign (positive or negative), and the charge The aggregates tend to break up easily under the

Suspensions
potential on the particle surface (including a double application of moderate shear stress, such as gentle

Super–
layer) govern the choice between flocculation or disper- agitation of a bottle or vial, or by the flow through a
sion (deflocculation). smallorifice (hypodermic needle and/or syringe) and
The processes involved in the formation of suspen- reform an extended network of particles after the
sions are shown in Fig. 2. The flocculated state (C) force is removed. Flocculation, therefore, imparts a
may be reached either directly by wetting and disper- structure to the suspension with virtually no increase
sing hydrophobic particles (A) with a suitable floccu- in viscosity (Fig. 3).
lating surfactant, or indirectly by first wetting and In contrast to peptized or deflocculated systems,
dispersing to produce a dispersed or peptized particle the stable floc settles rapidly, usually to a high sedi-
(B) with a suitable surfactant and then flocculating ment volume, and may be easily resuspended even after
with a suitable agent such as a hydrophilic colloid or standing for prolonged periods of storage time. The
polyelectrolyte. settling rate is not rapid and maintain content uni-
In contrast to peptized or deflocculated particles, formity with reasonable periodic agitation. The stable
flocculated suspensions (C), which are considered phar- floc can be produced by employing aseptic techniques,
maceutically stable (although colloidally unstable), using vehicle components that are safe for intramuscu-
can always be resuspended with gentle agitation. lar injection.
Severe overflocculation, on the other hand, caused Flocculated pharmaceutical suspensions are pre-
by the addition of too much flocculating agent or by pared using several methods. The choice depends on
prolonged exposure to extreme thermal conditions the properties of the drug and the class of suspension
(freeze–thaw cycles) tend to produce agglomerated or desired. The following examples illustrate how suspen-
coagulated irreversible systems (E). The term plaque sions may be prepared by controlled flocculation
(platelike) is used to describe essentially flat agglomer- procedures:
ates, whereas the term coagula (clumplike) is reserved
for thicker, three-dimensional particle masses. In the 1. The wetting agent, polysorbate 80 (not more
absence of a protective colloid, the process of crystal than 0.1–0.2% w/v of the final concentration),
growth is indicated by the arrow connecting (A) to (D). is dissolved in approximately half the final vol-
ume of aqueous vehicle. An anionic surfactant,
such as Docusate Sodium USP, may be used
Particle as a wetting agent. This agent is, however,
sensitive to pH and electrolyte concentration.
Boundary 2. Ultrafine particles of the drug at the desired
Layer
Wetting and final concentration are uniformily and carefully
1·10µ Range
Dispersion spread over the surface of the vehicle, and the
A B
Flocculation
Peptization or
Deflocculation
A B
Crystal
Growth Collodially Collodially Pharmaceutically
Agglomeration "stable" "unstable" "stable"
or Coagulation
Stable Floc
C Close packing Porous

Sediments slowly to Sediments rapidly to

> 10µ Range Agglomerate small sediment volume large sediment volume
or Coagulate difficult to redisperse easily redispersed
D E
Fig. 3 Characteristics of flocculated and deflocculated
Fig. 2 Processes involved in the formation of suspensions. suspensions. (From Ref.[5].)
 3604 Suspensions

drug is permitted to be wetted undisturbed for titrating concentrated aqueous solutions of soluble salt
as long as 16 h (overnight). forms of acidic or basic drugs with a corresponding
3. The wetted slurry is passed through a fine wire solution of a strong acid or a strong base. The water-
mesh screen (100 mesh size for ca 120 mm or insoluble free acid or base is precipitated at the pH
larger) to remove poorly wetted powder. A sin- of minimum solubility of the drug. The concentrations
gle pass through a colloid mill can achieve the of the reacting solutions and the order of addition may
same result. be varied to produce an acceptable stable floc. If neces-
Suspensions

4. The slurry concentrate of the drug is agitated sary, the electrolyte thus formed during precipitation
Super–

gently with an impeller-type mixer. may be reduced through slurry decantation or filtra-
5. Small amounts of a 10% w/v solution of alumi- tion to adjust tonicity and/or to maintain physical and
num chloride hexahydrate are added dropwise, chemical stability. This procedure can also be carried
to the drug slurry from a buret or dropping out under aseptic conditions.
pipette until the flocculation endpoint is reached A stable floc may also be produced by dispersing
(zero z-potential). To determine the endpoint, insoluble particles in a turbid or hazy vehicle consisting
small samples are withdrawn and transferred of finely dispersed or emulsified semipolar, liquid drop-
to a graduated cylinder, an equal amount of lets, which cause the droplets to be adsorbed on the
vehicle is added to each, and the cylinders are surface of the insoluble drug particles, resulting in a
gently shaken and permitted to stand undis- stable floc. Turbid aqueous vehicles have been pre-
turbed. The sample with the highest ratio of pared by the interaction of non-ionic surfactants and
sediment to total suspension volume, exhibiting preservatives. The concentration of surfactant and pre-
a clear supernate and good drainage character- servative required for haze formation may be reduced
istics is considered to be at the appropriate end- by the addition of small amounts of sorbitol to the
point. Usually, no more than about 0.1–0.2% vehicle.
aluminum chloride hexahydrate is be required to
achieve the flocculation endpoint. A 10% solu-
tion of calcium choride dihydrate may also be
Structured Vehicle
used as the flocculating agent. In this case, 1–2%
of the calcium salt may be required for stable
Another technique for the preparation of a stable sus-
floc formation. If the drug fails to flocculate in
pension is based on the concept of the ‘‘structured
the presence of polyvalent aluminum or calcium
vehicle,’’ in which the viscosity of the preparation,
ions, the water-insoluble drug particles are con-
under static conditions of very low shear, on storage
sidered to be positively charged, and the pro-
approaches infinity. The vehicle is said to behave like
cedure is repeated, this time using a polyvalent
a ‘‘false body’’ that is able to maintain the suspended
anionic flocculating agent such as 10% w/v
particles in a state of more or less permanent suspension.
sodium hexametaphosphate or 10% trisodium
Structured vehicles are usually not considered for
citrate.
the preparation of parenteral suspensions because,
6. After the flocculation endpoint has been estab-
owing to their high viscosity, such systems lack
lished and verified, the other components
sufficient syringe ability for ease of use.
(preservative, colorant, flavor, buffer, etc.) are
added, dissolved in the liquid vehicle, and the
slurry is brought to final volume with liquid
vehicle. Bingham-Type Plastic Flow

Another popular method of preparing an oral sus- Vehicles that exhibit the unusual property of Bingham-
pension consists of suspending the drug in a solution type plastic rheological flow are characterized by the
of a hydrophilic colloid (gelatin or gum) or a diluted need to overcome a finite yield stress before flow is
magma of bentonite, attapulgite, or colloidal magne- initiated. Permanent suspension of most pharmaceuti-
sium aluminum sililcate. The concentration of floccu- cal systems requires yield-stress values of at least
lating agent suspended in water, sorbitol, or syrup 2–5 Pa (20–50 dyn/cm2). Bingham plastic flow is rarely
solution is between 0.1 and 1%. Overflocculation may produced by pharmaceutical gums and hydrophilic
be reversed by the careful addition of small amounts colloids. National Formulary (NF) carbomers exhibit
of a suitable surfactant or polyvalent deflocculating a sufficiently high yield value at low solution con-
agent. Because clays cannot be used in injectable pro- centration and low viscosity to produce permanent
ducts, two other methods are described here. suspensions. The carbomers, however, require a pH
One method, specially useful for preparing physi- value between 6 and 8 for maximum suspension per-
cally stable ‘‘non-caking’’ suspensions, consists of formance. The polymer is essentially incompatible
Suspensions 3605

with cationic resins, certain polyvalent ions, and high systems is widely used in both pharmaceutical and
concentrations of electrolytes. cosmetic systems. A dilute emulsion system is not often
considered for suspension purposes because of the
potential complexities involved in mixing emulsion
Thixotropic Flow and suspension systems. The drug particles are dis-
persed in the primary emulsion component prior to
Thixotropy is a rheological property that results in dilution with other vehicle components. Emulsifiers

Suspensions
yield stress on standing. Thixotropic flow is defined that exhibit Bingham plastic or thixotropic flow char-

Super–
as a reversible, time-dependent, isothermal gel–sol acteristics should have acceptable formulating proper-
transition. Thixotropic systems exhibit easy flow at ties (taste, stability, etc.).
relatively high shear rates. However, when the shear
stress is removed, the system is slowly reformed into
a structured vehicle. The usual property of thixotropy
results from the breakdown and buildup of floccules FORMULATION OF SUSPENSIONS
under stress. A small amount of particle settling takes
place until the system develops a sufficiently high yield During the preparation of physically stable pharma-
value. The primary advantage of thixotropic flow is ceutical suspensions, a number of formulation compo-
that it confers pourability under shear stress and vis- nents are used to keep the solid particles in a state of
cosity and sufficiently high yield stress when the shear suspension (suspending agents), whereas other compo-
stress is removed at rest. nents are part of the liquid vehicle itself and have other
The concentrations of Bingham-type and thixo- functions in the dosage form.
tropic suspending agents required to achieve a yield
stress of 10 Pa (100 dyn/cm2) is shown in Table 2. 1. Components of the suspending system
Suspending agent systems such as a pseudoplastic
Wetting agents
(sodium carboxymethylcellulose) in combination with
Dispersants or deflocculating agents
a clay (hydrated colloidal magnesium aluminum sili-
Flocculating agents
cate) or blends and coprecipitates of sodium carboxy-
Thickeners
methylcellulose and microcrystalline cellulose exhibit
some thixotropic flow characteristics. Other pseudo-
2. Components of the suspending vehicle or exter-
plastics such as hydroxyethylcellulose or hydroxy-
nal phase
propyl methyl cellulose may be required to overcome
possible in compatibilities with sodium carboxymethyl- pH control agents and buffers
cellulose. Osmotic agents
Coloring agents, flavors, and fragrances
Preservatives to control microbial growth
Emulsion Base

An emulsion base or a waxy-type self-emulsifier to

develop structure or ‘‘false body’’ in suspension Wetting Agents

Wetting agents are surfactants that lower the inter-

Table 2 Concentration of suspending agents in water at facial tension and contact angle between solid particles
25
C required to achieve a yield value of 10 Pa and the liquid vehicle. When the insoluble powder is
Agent Concentration (%) added to a liquid vehicle containing a wetting agent,
Carbomer 941 0.1 penetration of the liquid phase into the powder will
be sufficiently rapid to permit air to escape from the
Carbomer 934 0.2
particles. The resulting wetted particles either sink en
Carrageenan 0.5 masse or separate with low-shear agitation. The best
Carboxymethylcellulose 2.0 range for wetting and spreading by non-ionic surfac-
Xanthan gum 2.0 tants is between a hydrophile–lipophile balance (HLB)
Algin 3.5 value of 7 and 10, although surfactants with values
Magnesium aluminum silicate 5.0 higher than 10 are often used for this purpose. Com-
mon wetting agents and surfactants include: 1) anionic
Hydroxyethylcellulose 5.0
type (docusate sodium and sodium lauryl sulfate) and
Guar gum 5.0 2) non-ionic type (polyoxyalkyl ethers, polyoxylakyl
Tragacanth gum 5.0 phenyl ethers, polyoxy hydrogenated castor oil,
(From B.F. Goodrich Specialty Chemicals, Cleveland, Ohio.) polyoxy sorbitan esters, and sorbitan esters).
 3606 Suspensions

Deflocculants and Dispersing Agents  Gums: acacia, agar, algins, carrageenan, guar,
pectin, tragacanth, xanthan
Unlike surfactants, these agents do not appreciably  Polymers: carbomers, polyvinyl alcohol, povidone,
lower surface and interfacial tension; thus, they have polyethylene oxide
little tendency to create foam or wet particles. Most  Sugars: dextrin, maltitol, sucrose
deflocculants, however, are not generally considered  Others: aluminum monostearate, emulsifying waxes,
safe for internal use, and as a result the only acceptable gelatin
Suspensions

dispersant for internal products is lecithin or a lecithin

Super–

derivative (naturally occurring mixture of phospha- The other agents used in pharmaceutical suspen-
tides and phospholipids). Because lecithins vary in sions (pH control agents, buffers, osmotic agents,
water solubility and dispersibility characteristics, stabilizers, vehicles, colorants, flavors, fragrances, and
proper control of product specifications must be main- preservatives to control microbial growth) are not
tained to obtain reproducibility. discussed in this article.

Flocculating Agents
STERILE SUSPENSIONS
Primary flocculating agents are simple neutral electro-
Sterile suspensions (injectable and ophthalmic) have
lytes in solution that are capable of reducing the z-
characteristics that are not commonly shared by other
potential of suspended charged particles to zero. Small
suspension systems, such as ease of resuspension, drain-
concentrations (0.01–1%) of neutral electrolytes, such
age, absence of foreign particulate matter and pyrogens,
as sodium or potassium chloride, are often sufficient
and syringeability in the case of injectable suspensions.
to induce flocculation of weakly charged, water-insolu-
The preparation of a sterile suspension is a difficult
ble, organic non-electrolytes. In the case of highly
manufacturing procedure. It requires strict attention
charged, insoluble polymers and polyelectrolyte spe-
to detail during the final recrystallization of the active
cies, similar concentrations (0.01–1%) of water-soluble
drug substance, size reduction, and sterilization of
divalent or trivalent ions, such as calcium salts, alums,
the active drug substance and the suspending vehicle,
sulfates, citrates, and phosphates, may be required for
aseptic wetting of the sterile drug powder with a por-
floc formation, depending on particle charge (positive
tion of the sterile vehicle, aseptic dispersion and milling
or negative). These salts are often used together as
of the bulk sterile suspension, and aseptic filling of the
pH buffers and flocculating agents.
finished suspension into sterile containers.
Various procedures for the manufacture of sterile
Thickeners, Protective Colloids, and suspensions have been reported by Akers et al.[12],
Suspending Agents Grimes,[13] and Portnoff.[14] At present, there is no phar-
maceutically acceptable chemical agent that can be
Protective or hydrophilic colloids, such as gelatin, added to the finished suspension to make it both sterile
natural gums, and cellulosic derivatives, that are and safe. Therefore, an elaborate program of sterility
adsorbed on insoluble particles, increase the strength checks at critical phases of the operation is required.
of the hydration layer formed around suspended parti- An important property of a good parenteral sus-
cles through hydrogen bonding and molecular interac- pension is syringeability; the ability of a parenteral
tion. Because these agents do not reduce surface and solution or suspension to pass easily through a
interfacial tension, they function best in the presence hypodermic needle, especially during the transfer of
of a wetting agent. Many of these agents are protective a product from vial to hypodermic syringe prior to
colloids in low concentration (<0.1%) and viscosity injection. Increases in vehicle viscosity, vehicle density,
builders in higher concentrations (>0.1%). Suspending and size and concentration of suspended particles
agents commonly used in pharmaceutical suspensions make the transfer more difficult.
include: The most important of these features is probably
viscosity. Fortunately, in the case of parenteral suspen-
 Cellulosics: sodium carboxymethylcellulose, micro- sions, viscosity is the easiest parameter to control.
crystalline cellulose (including coprecipitates and The preparation of a stable floc contributes little to
blends of the two), hydroxyethylcellulose, hydroxy- the overall viscosity of the system (the so-called Ein-
propyl cellulose, hydroxypropyl methylcellulose, stein relationship) and does not adversely influence syr-
methylcellulose, starch, sodium starch glycolate, ingeability. Although the individual particles are held
and powdered cellulose loosely together in large multiple aggregates, they are
 Clays: attapulgite, bentonite, magnesium aluminum easily broken up and reformed during the passage
silicate, kaolin, silicon dioxides from vial to syringe and syringe to injection site.
Suspensions 3607

Drainage, or the ability of the suspension to break Appearance

cleanly away from the inner walls of the primary
container-closure system, is another important charac- The appearance in a graduated glass cylinder or trans-
teristics of a well-formulated parenteral suspension. parent glass container is noted. The following ques-
Deflocculated to flocculated systems show this pro- tions were addressed: At equilibrium, is the color and
perty, whereas overflocculated systems show some appearance of the sediment uniform? Are there breaks
degree of poor drainage, also called ‘‘buttermilk or air pockets in the sediment? Is the residual drainage

Suspensions
appearance,’’ a term that aptly describes this unsightly above the sediment uniform and minimal, or is there

Super–
condition. Silicone coated containers, vials and plugs coagulated material adhering to the inside walls of
with dimethicone improve drainage and help reverse the container?
the tendency toward poor drainage by slightly over-
flocculated systems.
Photomicroscopic Examination
Resuspendability, or the ability to distribute settled
particles with a minimum of shaking, is an important
The microscope can be used to estimate and detect
characteristic of parenteral suspensions. Stable, floccu-
changes in particle size distribution and crystal shape.
lated parenteral suspensions that have been undisturbed
Its usefulness can be enhanced by attaching a Polaroid-
for long periods of storage time are easily resuspended.
type camera to the microscope to permit rapid pro-
cessing of photomicrographs (Fig. 4). These can be
used, for example, to distinguish between flocculated
COSMETIC SUSPENSIONS and non-flocculated particles and to determine changes
in the physical properties and stability. Sufficient fields
Cosmetic suspensions are available in two types. The and samples should be examined to make these deter-
first comprises pigmented products that are suspended minations. Dilutions for microscopic examination
in essentially aqueous vehicles (liquid makeup, eye- should be made with supernatant external phase rather
liners, mascara, and blusher). These products have a than with purified water. Individual particle size distri-
high solids content, high density, impalpable powders, butions can be accurately determined, using suitable
and pigments permanently suspended in a primary oil- electron instrumentation, for example, a Coulter
in-water emulsion-type base or a complex system of Multisizer II (an electrical sensing zone instrument
hydrophilic cellulose derivatives, clays, and/or poly- from Coulter Scientific Instruments, Hialeah, FL) or
meric film formers, in which the gelling and suspending the Elzone 280 PC systems (Particle Data, Elmhurst,
properties of the vehicle often are reinforced by a small IL). General methods for particle size analysis are
amount of a Bingham-type plastic such as carbomer. given in Fig. 5.
The second type comprises pigment-containing nail
enamels. The coloring tints, pigments, pearls, and lakes
are suspended with the aid of an organophilic, thixo- Color, Odor, and Taste
tropic gellant, such as stearalkonium hectorite in a
non-aqueous vehicle, containing butyl acetate, ethyl These characteristics are especially important in orally
acetate, and isopropyl alcohol solvents in which the administered suspensions. Variation in color often
primary plasticized nitrocellulose and toluene sulfon-
amide—formaldehyde resin film formers are dissolved.
Nail enamel is an excellent example of a permanent
suspension in a non-aqueous vehicle.

TEST METHODS FOR PHARMACEUTICAL

SUSPENSIONS

Tingstad[15] reviewed test methods for determining

the physical stability of pharmaceutical suspensions.
The procedures outlined are designed to determine
the state of flocculation of a formulation. Because
there is more than one method of preparing stable sus-
pensions, the following test methods and performance
criteria were found useful for determining the stability
of both flocculated and dispersed systems. Fig. 4 Photomicrograph of a flocculated steroid suspension.
 3608 Suspensions

Viscosity
Screening
A Brookfield viscometer with a helipath attachment
Aided sifting
(Stoughton, MA) is a useful rheological instrument
Sedimentation by gravitation
for measuring the settling behavior and structure of
pharmaceutical suspensions and for characterizing
the properties and stability of flocculated suspensions.
Suspensions

Sedimentation by centrifugal force

The viscometer should be properly calibrated to mea-
Super–

Optical methods using visible light

Electronic optical methods
sure the apparent viscosity of the suspension at equilib-
rium at a given temperature to establish suspension
Coulter reproducibility. Apparent viscosity, like pH, is an
counter
exponential term, and therefore the log-apparent vis-
10–1 1 101 102 103 104 105
cosity is an appropriate way of reporting the results.
Particle size in microns

Fig. 5 General methods for particle size analysis. Density

Specific gravity or density of the suspension is an

important parameter. A decrease in density often indi-
indicates poor distribution and/or differences in par- cates the presence of entrapped air within the structure
ticle size. Variations in taste, especially of active consti- of the suspension. Density measurements at a given
tuents, can often be attributed to changes in particle temperature should be made using well-mixed, uniform
size, crystal habit, and subsequent particle dissolution. suspensions; precision hydrometers facilitate such
Changes in color, odor, and taste can also indicate measurements.
chemical instability.

pH Value
Sedimentation Rate, Sediment Volume,
and Resuspendability The pH value of aqueous suspensions should be taken
at a given temperature and only after settling equilib-
Simple, inexpensive, graduated cylinders (100– rium has been reached, to minimize ‘‘pH drift’’ and
1000 mL) are useful for determining the physical stab- electrode surface coating with suspended particles.
ility of suspensions. They can be used to determine Electrolyte should not be added to the external phase of
the settling rates of flocculated and non-flocculated the suspension to stabilize the pH, because neutral elec-
suspensions and the sediment height at equilibrium. trolytes disturb the physical stability of the suspension.
The falling height of the liquid—sediment interface
of the suspension is determined as a function of time,
and the sedimentation rate test is repeated periodically Freeze—Thaw Cycling
during storage. The sediment volume at equilibrium
should be sufficiently large to support uniform resus- This is a useful guide to the physical stability of sus-
pension with gentle agitation. The equilibrium sedi- pensions. If freeze—thaw cycling or elevated tempera-
ment volume should be similar and reproducible ture exposures are chosen for physical stability testing,
batch after batch. companion samples of a closely related marketed sus-
Volumetric graduated cylinders are used to deter- pension should be included in the testing protocol for
mine the ‘‘F’’ or flocculation ratio, a value that repre- comparison purpose because pharmaceutical suspen-
sents the ratio of the sediment volume to the original sions are not normally designed to withstand tempera-
suspension volume at a given time. It is used to mea- ture extremes during storage (15–30
C optimum).
sure the relative degree of flocculation and physical
stability of suspensions. A sufficiently wide graduated
cylinder should be used to overcome the ‘‘wall effect’’ Drug Content Uniformity
which often influences the settling rate and equilibrium
sediment volume of flocculated suspensions. Small This important testing procedure is best performed using
graduated cylinders have a tendency to support sus- either ‘‘unit of use’’ volume (e.g., 5 mL of oral liquid or a
pensions because of the adhesive forces acting between spray actuation of an oral inhalation product) or sam-
the inner surface of the container and the suspended pling from a well-mixed dispensing container from the
particles. top, middle, and bottom of the suspension.
Suspensions 3609

Dissolution Testing The techniques used for preparing nanoparticles are

similar to those used to prepare more conventional
This technique[16] is still evolving. The favored approach drug particles and include controlled precipitation,
at present is to submerge a small, known amount of ball milling using glass or zirconium oxide pearls, and
suspension inside a secured Durapore (polyvinylidene high-pressure homogenization.
fluoride) membrane pouch (Millipore Products, Nanosuspensions can be sterilized for parenteral
Bedford, MA) of suitable porosity in ‘‘teabag’’ fashion use by using conventional steam sterilization in an

Suspensions
in a suitable dissolution medium using the USP Method autoclave, g-irradiation, or membrane microfiltration

Super–
1 Paddle Apparatus.[17] Optimization of experimental in certain situations.
conditions (rate of agitation, volume and type of The key to long-term physical stability of aqueous
medium, temperature, etc.) must be established to nanosuspensions is the selection of a suitable water-
achieve reproducible results. soluble surfactant or polymer as an external particle
stabilizer to prevent particle growth. Several potential
stabilizers are lecithin, phospholipids, poloxamers,
Particle Size Measurement and polysorbates.
The physical stability of nanosuspensions may be
Recently,[18–20] with respect to the importance of monitored with the use of electron microscopic analysis.
particle size distribution in terms of particle character- The major advantage of pharmaceutical nanosus-
ization and product physical stability testing, there has pensions is their ability to increase the in vivo absorp-
been interest in newer light-scattering methods for tion of highly–water-insoluble drugs by dramatically
particle detection called photon correlation spectro- reduced particle size.
scopy (PCS). PCS methods can be applied to both
micro-and nanosuspensions.
The information thus obtained from the use of such
REFERENCES
equipment includes mean particle size, particle size dis-
tribution, particle concentration, molecular weight esti- 1. Martin, A.; Bustamante, P. Physical Pharmacy, 4th Ed.;
mation, polydispersity, particle shape, hydrodynamic Lea & Febiger: Philadelphia, 1993; 477–484.
interactions, and aggregation mechanisms. 2. Marcus, A.D.; DeKay, H.G. New developments in cala-
mine lotion. J. Am. Pharm. Assoc., Pract. Ed. 1950, 11,
In addition,[21] there are several experimental options 227–229.
available for particle size measurement alone. They 3. Boylan, J.C. The development of semi-solid dosage forms:
include single particle optical sensing (SPOS), laser an overview. Drug Dev. Commun. 1976, 2, 320.
4. Robinson, M.J. Third Annual National Industrial Pharma-
diffraction (LD), and ultrasound attenuation (UA). ceutical Research Conference, Land o’lakes. WI, 1961.
5. Hiestand, E.N. Theory of coarse suspension formulation.
J. Pharm. Sci. 1964, 53, 1–18.
6. Carpenter, C.R. Calculate settling velocities for unrestricted
Other Procedures particles on hindered settling. Chem. Engr. Nov. 1983, 4.
7. Haines, B.A.; Martin, A.N. Interfacial properties of
Assays for potency, preservative effectiveness, com- powdered material. J. Pharm. Sci. 1961, 50, 228–232,
753–759.
patibility with primary container-closure system, off- 8. Matthews, B.A.; Rhodes, C.T. Use of DLVO theory to
torque, simulated use testing, etc., should be handled interpret pharmaceutical suspension stability. J. Pharm.
in a manner similar to that used for conventional Sci. 1970, 59, 521–525.
9. Ecanow, B.; Gold, B.; Ecanow, C. Newer aspects of
liquid solutions, with the provision that the container suspension theory. Am. Perfum. Cosmet. Nov. 1969, 84,
is well-mixed prior to sampling. 27–31.
10. Ecanow, B.; Webster, J.; Blake, M.I. Conductivity studies
of suspension systems in different states of aggregation.
J. Pharm. Sci. 1982, 71, 456–457.
11. Michaels, A.S.; Bolger, J.C. The plastic flow behavior of
NANOSUSPENSIONS flocculated kaolin suspension. Ind. Eng. Chem. Fundam.
1962, 1, 153–162.
12. Akers, M.J.; Fites, A.L.; Rabinson, R.L. Formulation
Pharmaceutical nanosuspensions[22] are usually very design and development of parenteral suspensions. J.
finely dispersed solid drug particles in an aqueous Parenter. Sci. Technol. 1987, 41, 88–96.
vehicle for either oral and topical use or for parenteral 13. Grimes,, T.L. Scaleup and Manufacture of Sterile Suspen-
sions. APhA 133rd Annual Meeting, San Francisco,
and pulmonary administration. The key difference March 19, 1986.
from conventional suspensions is that the particle size 14. Portnoff, J.B. The Development of Sterile Suspensions—
distribution of the solid particles in nanosuspensions Case Study. APhA 133rd Annual Meeting, San Francisco,
March 19, 1986.
is usually less than 1 mm, with an average particle size 15. Tingstad, J.E. Physical stability testing of pharmaceuticals.
range between 200 and 600 nm. J. Pharm. Sci. 1964, 53, 955–962.
 3610 Suspensions

16. Stout, P.J.; Howard, S.A.; Mauger, J.W. Dissolution of phar- BIBLIOGRAPHY
maceutical suspensions. In Encyclopedia of Pharmaceutical
Technology; Swarbrick, J., Boylan, J.C., Eds.; Marcel Dekker,
Banker, G.S.; Rhodes, C.T. Modern Pharmaceutics, 2nd Ed.;
Inc.: New York, 1991; Vol. 4, 169–192.
17. United States Pharmacopeia 24, The Pharmacopeial Con- Rev. Exp.; Marcel Dekker, Inc.: New York, 1989; 339–353.
vention, Rockville, MD 2000, 4. Handbook of Pharmaceutical Excipients, 3rd Ed.; APhA and
18. Castanho, M. Light Scattering and Photon Correlation Pharmaceutical Press: London, 2000.
Spectroscopy; NATO ASI Series; High Technology, 1997; Martin, A.; Bustamante, P. Physical Pharmacy, 4th Ed.; Lea &
40, 31–36. Febiger: Philadelphia, 1993; 477–484.
Suspensions

19. Constantinides, P.P.; Yiv, S.H. Int. J. Pharm. 1995, 115, 225. Nash, R.A. Pharmaceutical suspensions. In Pharmaceutical
20. Ruth, H., Saint. Int. J. Pharm. 1994, 116, 253. Dosage Forms, Dispersed Systems; Lieberman, H.L.,
Super–

21. Nicoli, D. Am. Lab. Jan. 2000. Rieger, M.M., Banker, G.S., Eds.; Marcel Dekker, Inc.:
22. Miiller, R.H.; Jacobs, C.; Kayser, O. Nanosuspensions. In New York, 1988; 1, 151–198.
Pharmaceutical Emulsions and Suspensions; Nielloud, F., Zografi, G.; Swarbrick, J.; Schott, H. Disperse systems. In
Marti-Mestres, G., Eds.; Marcel Dekker, Inc.: New York, Remington’s Pharmaceutical Sciences; Gennaro, A.R., Ed.;
2000. Mack Publishing Co.: Easton, PA, 1990; 257–309.
Tablet Compression: Machine Theory, Design,
and Process Troubleshooting
Michael J. Bogda
Barr Laboratories, Inc., Woodcliff Lake, New Jersey, U.S.A.

INTRODUCTION  Improved force-measurement techniques.

 Introduction of electronics to provide force control.
The most common method of drug delivery is the oral  Integration of on-line weight, thickness, and hard-

Tablet–Tablet
solid dosage form, of which tablets and capsules are ness test units providing weight feedback control
predominant. The tablet is more widely accepted and to the force control unit, and
used compared to capsules for a number of reasons,  High-speed single-tablet sorting to reject out-of-
such as cost, tamper resistance, ease of handling and specification tablets.
packaging, ease of identification, and manufacturing
efficiency. Over the past several years, the issue of Therefore, optimal product development can
tamper resistance has resulted in the conversion of most typically be performed on these machines that offer
over-the-counter drugs from capsules to predominantly improved compression designs and material feed
all tablets. systems.
Pharmaceutical products have been manufactured This article provides the basic information neces-
into compressed tablets for many years. During the sary to understand the general process of tablet forma-
1950s, much research was devoted to the physics of tion. General machine design characteristics and tablet
compression.[1,2] Since that time, the pharmaceutical press nomenclature are presented. Tablet press control
industry has attained a much greater understanding systems and process automation are discussed,
of the compression process, which resulted in the followed by process and product troubleshooting on
development of more robust pharmaceutical formula- tablet compression equipment.
tions.[3–5] This has been achieved by the use of
instrumented tablet presses and sophisticated data col-
lection systems combined with the development of THE PROCESS OF TABLET FORMATION
mathematical models.
During this time, a significant portion of the devel- The quality of a compressed tablet is determined by
opment work has been conducted on older equipment, material fill characteristics and compression behavior.
which has been retrofitted to measure compression During compression, the rate at which tablets can be
and ejection-force signals. Recent advances in the produced can be limited due to non-uniform material
design of tablet compression equipment has resulted fill characteristics. Pending successful and reproducible
in higher-efficiency machines designed to optimize material fill (die fill), the powder mass must form a
compression efficiency, minimize tablet weight vari- coherent compact that stays intact upon ejection out
ation, and provide greater flexibility, allowing the pro- of the die. Therefore, tablet press performance can be
duction of a greater range of products. However, the limited due to poor fill characteristics and/or poor
modern sophisticated machines still employ the same compression behavior.
general concepts of operation: die fill, tablet com- Die fill characteristics depend upon material flow
pression, tablet ejection, and tablet scrape-off. There- properties that are primarily affected by particle size
fore, studies conducted on older equipment designed and shape. Additionally, high interparticle friction
to evaluate the compression characteristics of mate- can have a detrimental effect on die fill characteristics
rials, can offer significant insight into material behavior. due to powder bridging and non-uniform flow charac-
However, modern machines provide greater accuracy teristics. A non-uniform particle size distribution may
and efficiency as follows: also lead to material segregation resulting in unifor-
mity problems. Tablet presses employ volumetric filling
 Improved material feed systems. of the material into the die cavity. Most high-speed
 Improved cam design and material of construction. tablet presses are equipped with force feeders, which
 Multistage compression. use rotating paddles to promote uniform die fill char-
 Isolated design for quick cleaning and changeover. acteristics. For certain materials, attention must be
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-100200019
Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved. 3611
 3612 Tablet Compression: Machine Theory, Design, and Process Troubleshooting

paid to ensure that the action of the power feeder does deformation may become plastic, that is, the
not cause overmixing or segregation of the material. particles undergo viscous flow. This is the pre-
Upon proper die fill, one must consider whether the dominant mechanism when the shear strength
material will form a tablet. The tableting characteris- between the particles is less than the tensile
tics of powders depend on the viscoelastic properties or breaking strength. Plastic deformation is a
of the material. The process of compaction has been time-dependent process.
defined as ‘‘the compression and consolidation of 3. Brittle fracture: Upon exceeding the elastic
a two-phase system due to an applied load.’’[6] The limit of the material (yield point), the particles
quality of the compact depends on the compression undergo brittle fracture if the shear strength
and consolidation of the powder mass, decompression between the particles is greater than the tensile
of the compact, ejection from the die, and subsequent or breaking strength. Under these conditions,
scrape-off from the lower punch. A schematic rep- the larger particles are sheared and broken into
resentation of the compression process is shown in smaller particles.
Fig. 1. Because these viscoelastic properties are time
Tablet–Tablet

dependent, both the magnitude and the rate of appli- The compression process includes these three
cation (and release) of the compression force affect mechanisms. The individual characteristics of the
tablet quality. material under investigation determine the extent to
which each is active. Since some of these deformation
characteristics are time-dependent, machine character-
Compression and Consolidation istics can have a major effect on tableting performance.
These characteristics determine the rate of force appli-
During compression, the bulk volume of the material is cation, dwell time (i.e., the time of maximum com-
reduced, resulting in the displacement of the gaseous pression force, which depends on the punch head flat
phase (air).[6] Further increasing the force leads to par- diameter and the tangential velocity), and the rate of
ticle deformation and rearrangement. At this point, the decompression (Fig. 1).
three principal modes of deformation are as follows: Typically for materials that undergo plastic defor-
mation, as machine speed is increased there is less time
1. Elastic deformation: A spontaneously reversi- for stress relaxation. Under these conditions, the
ble deformation of the compact in which, upon tablets may cap and laminate. However, capping and
removal of the load, the powder mass reverts lamination can be eliminated or minimized by slowing
back to its original form. Most materials down the compression process (reducing the machine
undergo elastic deformation to some extent. speed), lowering the rate of force application (larger
Compression of rubber would be by elastic compression roller diameter), or increasing the time
deformation. of compression (multistage compression).
2. Plastic deformation: After exceeding the The final tablet properties are also affected by the
elastic limit of the material (yield point), the consolidation (i.e., bonding) mechanisms of the
powder which is influenced by its chemical nature,
the surface area of the contact points, contamination
(including film coatings such as magnesium stearate),
Compression and interparticle distance. The predominant consoli-
Roller
dation mechanisms are listed below:[7–10]

 Mechanical theory: As the particles undergo

deformation, the particle boundaries intertwine to
form mechanical bonds.
 Intermolecular forces theory: van der Waals forces
Force

bond the molecules together at the newly sheared

surfaces of the particle boundaries. Microcrystalline
cellulose is believed to undergo significant hydrogen
bonding during tablet compression.[11]
 Liquid-surface film theory: Thin liquid films form
Compression which bond the particles together at the particle
& Consolidation Decompression Ejection Scrape-off
surface. The energy of compression produces
Time
melting or solution at the particle interface followed
by subsequent solidification or crystallization thus
Fig. 1 The compression process. resulting in the formation of bonded surfaces.
Tablet Compression: Machine Theory, Design, and Process Troubleshooting 3613

Due to the applied pressure, the particles may melt may improve the tableting characteristics more than
or dissolve. As the pressure is released, solidification conventional tableting where main compression
and crystallization occur. exceeds precompression. It is theorized that this effect
is due to the high initial compression force that raises
The intermolecular forces theory and the liquid- the tablet temperature, thereby increasing ductility
surface film theory are believed to be the major and allowing greater plastic deformation. Application
bonding mechanisms in tablet compression. Many of the lower second compression force increases the
pharmaceutical formulations require a certain level of formation of particle–particle bonds while minimizing
residual moisture to produce high quality tablets. The particle-bond rupture, resulting in stronger tablets.
role of moisture in the tableting process is supported
by the liquid-surface film theory.
During tablet formation, as load is applied to the Decompression
powder mass, plastic deformation and brittle fracture
create clean surfaces that are brought in intimate con- After the compression and consolidation of the powder

Tablet–Tablet
tact by the applied load. These surfaces bond together in the die, the formed compact must be capable of
by a number of mechanisms such as those listed earlier. withstanding the stresses encountered during decom-
Plastic deformation is believed to create the greatest pression and tablet ejection.[19] The rate at which the
number of clean surfaces. Because it is time-dependent, force is removed (dependent on the compression roller
higher rates of force application should lead to the for- diameter and the machine speed) can have a significant
mation of less new clean surfaces, resulting in weaker effect on tablet quality. The same deformation charac-
tablets. Additionally, because tablet formation is teristics that come into play during compression play a
dependent upon the formation of clean new surfaces, role during decompression.
high concentrations or overmixing of materials that After application of the maximum compression
form weak bonds result in weak tablets. Magnesium force, the tablet undergoes elastic recovery. While the
stearate, for example, forms weak bonds and easily tablet is constrained in the die, elastic recovery occurs
wets surfaces. Therefore, overlubrication or overmix- only in the axial direction. If the rate and degree of
ing of magnesium stearate may lead to weak tablets. elastic recovery are high, the tablet may cap or lami-
Fragmentation (the creation of new clean surfaces) nate in the die due to rapid expansion in the radial
continues at the same time at which bonding and den- direction only. If the tablet undergoes brittle fracture
sification occur. A high quality tablet can be formed during decompression, the compact may form
only when the process of bonding and densification failure plains due to the fracturing of the surfaces.
exceeds fragmentation. The rates at which these func- Tablets that do not cap or laminate are able to relieve
tions occur are dependent upon the rate at which the the developed stresses by plastic deformation. Since
forces are applied. plastic deformation is time-dependent, stress relaxation
During compression, the powder compact typically is also time-dependent. Therefore, tablet fracture is
undergoes a temperature increase,[12,13] which depends affected by rates of decompression (machine speed)
on frictional effects that are dependent, in turn, on the since the compact may not have sufficient time to
specific material characteristics, the lubrication efficiency, relieve the internal stresses created during decom-
the magnitude and rate of application of the compression pression. Formulations which contain significant con-
forces, and the machine speed. Typical temperature centrations of microcrystalline cellulose typically
increases are between 4 and 30 C.[14] As the tablet tem- form good compacts due to its plastic deformation
perature rises, stress relaxation and plasticity increase, properties. However, if the machine speed and the
while the elasticity decreases and strong tablets are rate of tablet compression are significantly increased,
formed.[15] Therefore, compression of material, at ele- these formulations exhibit capping and lamination
vated temperature with increased ductility should result tendencies.
in stronger tablets. The rate of decompression can also have an effect
It is believed that under certain conditions precom- on the ability of the compacts to consolidate (form
pression is beneficial because it helps to remove bonds). Based on the liquid-surface film theory, the
entrapped air[16] and extends dwell time, thereby rate of crystallization or solidification should have an
increasing the degree of stress relaxation and plastic effect on the strength of the bonded surfaces. The rate
deformation[17] as well as the number of bonds, thus of crystallization is affected by the pressure (and the
increasing tablet strength. Additionally, by extending rate at which the pressure is removed). High decom-
the time of compression, precompression may provide pression rates should result in high rates of crystalliza-
a gradual loading and unloading of force. However, tion. Typically, slower crystallization rates result in
recent studies[18] suggest that a high level of precom- stronger crystals. Therefore, if bonding occurs by these
pression (higher than that of the main compression) mechanisms, lower machine speeds (lower rates of
 3614 Tablet Compression: Machine Theory, Design, and Process Troubleshooting

decompression and crystallization) should result in Standard Stripper

stronger tablets.

Ejection

After decompression, the tablet remains in the die until

it is ejected. During this time, a residual die wall force
is exerted by the tablet on the die wall. Tablet ejection
is defined by three stages:[6] Modified
Stripper

1. The initial ejection peak force required to break

the tablet adhesion to the die wall. This force is
the highest force encountered during ejection
Tablet–Tablet

and occurs over a very short period of time.

2. The force required to push the tablet up the die
wall, which is typically lower than the ejection Fig. 2 Tablet stripper.
peak force. However, inadequate lubrication
or damaged dies may result in slip-stick behav-
ior where the tablet continues to adhere and At high machine speeds, scrape-off problems may
break adhesions to the die wall surface. These be encountered due to tablets backing up at the point
conditions typically result in tablet failure. of discharge. Under many circumstances, specially
3. Declining forces as the tablet emerges from the die. modified tablet strippers have proven to be beneficial
for shaped tablets by discharging the tablets off the
Inadequate lubrication typically results in high ejec- die table as quickly as possible (Fig. 2).
tion forces and possibly tablet failure. Ejection forces
below 200 N are optimal, although forces up to
400 N are common. Ejection forces ranging between
400 and 800 N are considered high. Forces exceeding ROTARY TABLET PRESS DESIGN
800 N result in excessive heat build-up and could lead
to machine damage and product failure. Inadequate Pharmaceutical tablets are generally produced on
lubrication can also result in striations along the tablet rotary tablet presses (Fig. 3), where upper and lower
side wall, and picking and sticking. punches reside in the upper and lower turret, respect-
Tablets that undergo significant elastic recovery ively. The dies are inserted in the die table and secured
upon decompression may exhibit capping upon ejec- by die lock screws. The upper and lower turret and the
tion out of the die. Under these circumstances, the tab- die table are precisely aligned. The movement of the
let builds up significant stress while in the die, which
can only be relieved in the axial direction. As the tablet
Direction of Rotation
emerges from the die, its top portion can expand in
both the radial and axial directions while the portion
remaining in the die is confined. Shear stresses develop Die Table
along the edge of the die and result in tablet failure. Recirculation
Channel
Tablet
Stripper
Scrape-Off

Tablet scrape-off occurs immediately after ejection.

Fig. 2 illustrates a tablet stripper on a rotary tablet Excess
Material
press. Typical forces during tablet scrape-off are 2 N Feed Frame Stripper
or less. Scrape-off forces of 6 N or higher result in
tablets sticking to the lower punches and subsequently
picking or, under extreme circumstances, shearing the
bottom of the tablet. Frequently, shearing of the lower
portions of the tablet due to scrape-off problems is
mistaken for capping. However, this can be easily
distinguished by examining the lower punches and
rotating the press manually. Fig. 3 Rotary tablet press (top view).
Tablet Compression: Machine Theory, Design, and Process Troubleshooting 3615

punches is controlled by cam tracks and compression depths, whereas variable granulation density within a
rollers. As the entire assembly rotates, the upper and batch results in fluctuating fill depth requirements.
lower punches move along the cam tracks to accomplish As the lower punch passes from the fill cam to the
die fill, tablet compression, ejection, and scrape-off. weight-regulation cam, the excess material is pushed
back into the feeder and scraped off at the top of the
die table by the excess material stripper and directed
Tablet Compression into a recirculation channel. On many modern presses,
the lower punch is lowered by approximately 2–4 mm
Tablet compression can be separated into the two dis- relative to the top of the die table after the excess
tinct yet equally important phases of die fill-weight material stripper. This lowers the material away
adjustment and tablet formation as shown in Fig. 4. from the top of the die table, minimizing uncontrolled
As die fill begins, the lower punch face is initially flush loss as the upper punch enters into the die cavity
with the die table surface as the lower punch enters the after scrape-off. Under these circumstances the upper
overfill cam at the entry of the feeder. The lower punch punch does not contact the top of the material until

Tablet–Tablet
travels under the feeder and is pulled down by the it enters into the die, minimizing material loss and
overfill cam. At this point the lower punch has passed weight variation. Additionally, lowering the slug of
through approximately 50% of the feeder and the die material away from the die table surface reduces
cavity contains more material than required. material loss due to the centrifugal force of the rotating
After overfilling the die cavity, the lower punch is die table.
adjusted to a constant height as it passes into the Overfilling of the die cavity is necessary to achieve
weight-regulation unit. The constant height, known uniform tablet weights and to optimize machine run-
as the fill depth, is measured as the distance between ning conditions. However, at times excessive overfilling
the lower punch face and the die table surface. Since can lead to other problems such as excessive wear for
die fill is volumetric, the constant height of the lower abrasive raw materials, particle size reduction for fri-
punch in the weight-regulation unit provides a constant able granulations, material segregation, and overmix-
volume of material. Therefore, the fill depth is affected ing of lubricant. Therefore, the effect of different
by the density of the granulation. Variation of granu- machine running conditions must be evaluated for
lation density between batches results in different fill each different product separately.

Start of
Cycle
Main
Precompression Compression
Die Weight Ejection
Fill Adjustment and
Position Position Scrape-Off
Position

Fill Weight
Ejection
Cam Regulation
Cam
Unit
Direction of Rotation

Fig. 4 Rotary press (sequences of compression).

 3616 Tablet Compression: Machine Theory, Design, and Process Troubleshooting

The material that is directed into the recirculation to provide separation and isolation of the compression
channel is subsequently introduced back into the area from the other components are as follows:
feeder at the tablet stripper or at the inside edge of
the feeder. It is worth noting that the paddle in the fee-  Upper cam section
der at the point of material entry rotates in the  Compression section
opposite direction as the turret to aid in die fill and  Lower cam section
induce flow back into the feeder.  Lower mechanical section
Frequently the maximum machine speed may  Electrical section
depend on die fill characteristics due to excess tablet
weight variation at high machine speeds. However, With the proper separation of these areas, only the
because the compression characteristics of most phar- compression zone is exposed to material, thus reducing
maceutical products exhibit viscoelastic properties, cleaning and change-over time of the tablet press. In
the press speed may also have a major effect on the addition to the machine sections, an understanding
compressibility of the material. For this reason, of other machine subsystems is necessary, such as the
Tablet–Tablet

the zability to compress a tablet adequately is often lubrication system and the diagnostic systems (safety
the overriding factor to consider in tablet compression. systems) to achieve optimal machine performance.
The process of tablet formation begins as the upper Modern rotary tablet presses are either single-sided
punch is lowered directly into the die cavity after the or double-sided. A single-sided machine has one feed-
excess material stripper. As mentioned previously, it ing station, one set of precompression and main com-
is advantageous if the slug of material is lower than pression rollers, and one discharge station. These
the die table surface as the upper punch enters to mini- machines produce one tablet per punch station per
mize uncontrolled material loss and weight variation. die table revolution. A double-sided machine has two
After the upper punch enters into the die, the upper feeding stations, two sets of precompression and main
and lower punches begin to move toward each other compression rollers, and two discharge stations, and
as the punches ride along cam tracks toward the pre- produces two tablets per punch station per die table
compression rollers. At the precompression stage, the revolution. The double-sided machine operates identi-
initial (and typically the lower) compression force is cally to the single-sided machine with the exception
applied. Traditionally, tablet presses were equipped that the excess material from the first feeding station
to apply a 20 kN maximum precompression force using passes into the second feeding station. A double-sided
relatively small compression rollers (approximately machine has a higher output than a single-sided
100-mm (4-in.) diameter rollers). However, to improve machine. Its pitch circle diameter is also greater, which
flexibility, many modern rotary tablet presses are could result in weight uniformity and compressibility
equipped with identical precompression and main issues.
compression force capabilities, allowing the application Bilayer tablet presses employ the same general
of 80–100 kN forces using 250–300-mm-diameter com- design concepts as single-sided machines. Typically, a
pression rollers. double-sided machine can be converted to a bilayer
After application of the precompression force, the machine by replacing various cams. The material for
punches move toward the main compression rollers each layer is introduced separately into each feeder
where the final (main) force is applied. As the punches and is removed from the die table to prevent contami-
impact the rollers, the compression force increases nation.
until the punch head flat is tangent to the compression
roller and maximum force is applied (Fig. 1). The
applied compression force is a measured value and Upper cam section
depends on the distance between the punches and the
quantity of material in the die. After main com- The upper cam section is typically shrouded and sealed
pression, the upper punch is pulled out of the die cavity to prevent exposure of material. It consists of the upper
while the lower punch impacts the ejection cam to cam track, all upper compression rollers, and all
begin the ejection process. As the die table continues adjustments to the position of the upper compression
to rotate, the lower punch raises the tablet out of the rollers. The primary components of the upper cam sec-
die cavity to eject the tablet to the point of scrape-off. tion are as follows:

1. Upper punch removal/dwell cams: The upper

Press Design and Layout punches are loaded and removed from the
machine at this location. These cams typically
Modern rotary tablet presses are typically designed in reside directly above the material feeder. In
separate machine sections (press zones). Typical sections many press designs, the upper punch dwell
Tablet Compression: Machine Theory, Design, and Process Troubleshooting 3617

cam is designed to measure the tightness of the

upper punches in the turret. A spring loaded
cam designed to raise the upper punch slightly
(1–4 mm) is connected to a proximity sensor.
If the punches are too tight then the spring-
loaded cam falls instead of raising the upper Adjustment
of
punches, thus tripping the proximity sensor
Insertion Depth
and shutting down the machine. In alternative
press designs, the upper punch tightness is mea-
sured in the upper-punch pull-up cam, typically
by a strain gauge measurement of the lifting
force.
2. Upper punch lowering cam: The upper punches Edge Insertion
are lowered into the die cavity by the upper- Thickness

Tablet–Tablet
Depth
punch lowering cam. This cam is typically
CAD optimized to minimize the acceleration
and velocity of the upper punch as it enters into
the die cavity. In this way, the upper punch tra-
vels in a smooth and controlled manner as it
enters the die cavity, thus improving weight
Adjustment
uniformity. of
3. Upper precompression and main compression Edge Thickness
rollers insertion depth adjustments: Insertion
depth for both precompression and main com-
pression is adjusted in the upper cam section.
The insertion depth determines the location of
tablet formation in the die cavity relative to
the top of the die table as shown in Fig. 5. It Fig. 5 Rotary tablet press layout.
is measured as the distance at which the upper
punch enters into the die at the tangent between
the upper punch head and the compression tight punches at this location prevents almost all
roller. Insertion depth can be varied between 2 possibility of machine damage.
and 6 mm on most machines and is typically 5. Cam material of construction: Both the upper
maintained between 3 and 4 mm. For precom- and lower cam sections use cams to guide the
pression and main compression, the insertion punches while the turret rotates. These cams
depth should be maintained at approximately are typically made of various materials such as
the same position. On most modern rotary tab- steel, bronze, or alloy. Most of the cam tracks
let presses, the adjustments for precompression in the turret are designed to smoothly guide
and main compression insertion depth are inde- the punches. Many modern rotary tablet presses
pendent. However, on many older designs, the use polymer composite cams for non-impact
precompression roller is attached to the main points. These cams have excellent qualities in
compression roller assembly and its position is that they provide superior abrasion resistance
measured relative to the main compression and have self-lubricating properties minimizing
roller position. In this way, the ratio of precom- cam and tool wear, heat generation, and noise,
pression to main compression remains constant and ultimately resulting in increased machine
as machine adjustments are made. speeds. However, cams that undergo impact
4. Upper punch pull-up cam: After compression, (e.g., ejection cam) and stress (e.g., weight-
the upper punch enters into the upper-punch regulation cam) require metal construction with
pull-up cam, which removes the upper punch good impact resistance. For this purpose, an
from the die cavity. This cam provides an excel- aluminum–bronze alloy provides superior abra-
lent location to measure the upper punch pull- sion resistance and excellent impact strength.
up force that determines the tightness of the
upper punches. Compared to the upper punch Compression section
dwell cam, this location has the advantage of
determining the punch tightness not only in The compression section contains all components that
the turret but also in the die cavity. Detection of are exposed to the material, such as the material
 3618 Tablet Compression: Machine Theory, Design, and Process Troubleshooting

hopper, the feeder, the excess material stripper, the turret rotational forces to achieve die fill. These feed
upper and lower turrets, the die table, and the tablet frames provide good performance for materials with
stripper. Additionally, the dust-collection shrouds are good flow properties but are typically limited to slow
located in the compression section. Proper shrouding machine speeds. On the other hand, gravity feeders
of this area ensures that none of the upper and lower do not agitate the product and impart no energy.
punch heads, compression rollers, and cam tracks are Therefore, they offer advantages for products where
exposed to material. Proper maintenance and setup material segregation and overmixing are of concern.
of the compression section is critical for optimal press For example, products that are sensitive to overblend-
performance. The primary components of the com- ing of magnesium stearate (i.e., exhibit capping when
pression section are explained in the following section: overblended) may exhibit improved compressibility
by using a gravity feeder as opposed to a force feeder.
Material Hopper. The material hopper is an integral
part of the feeding system as shown in Fig. 6. Force Feeder. Force feeders are typically multichamber
and multipaddle feeders as shown in Fig. 6. These feeders
Tablet–Tablet

Typically, it is capable of holding approximately

5–10 kg of material. Low level sensors are mounted are critical to allow optimal press performance at high
in the hopper to signal an alarm, shut off the machine machine speeds with minimal weight variation. For pro-
or activate a feeding mechanism to deliver more ducts with good flow properties, the feeder should move
material when the product falls below this level. The the material from the overhead hopper to the dies with
material hopper should be symmetrical with steep dis- minimal mixing. Most force feeders contain two or three
charge angles to promote mass flow and prevent funnel chambers and paddles. The three chamber/paddle sys-
flow (rat holing) in the granulation. The discharge out- tem typically performs better than the two chamber/
let of the hopper should be as large as possible reach- paddle designs. The top paddle and feed chamber are
ing into the feeder to prevent material bridging. connected directly to the hopper and move the material
On many machines the base of the hopper is equipped from the overhead hopper to the filling chambers located
with a valve to shut off material flow to the feeder if directly above the die cavities. The top chamber elimi-
necessary. Depending on the nature of the granulation, nates the effect of the head pressure on material flow,
the hopper valve can contribute to material bridging. thus providing uniform die fill regardless of the quantity
For materials with very poor flow characteristics, a slide of material in the hopper. Alternate systems offer level
valve may be preferable to a butterfly valve. sensors that are designed to provide a constant quantity
Older generation machines typically employ large of material to the feeding chambers, thus also eliminating
hoppers, but more recent designs offer options to the effect of head pressure on material flow.
install only a straight chute in place of a large overhead The force feeder chambers contain material baffles
hopper. This design minimizes poor flow behavior and that function to prevent the material from randomly
segregation due to funnel flow. packing in the chambers, which results in non-uniform
fill. Optimal systems provide minimal energy input and
minimal particle mixing while providing uniform fill.
Gravity Feed Frame. Older machines typically employ
The speed of the paddles can be sychronized with the
gravity feed frames which rely on gravitational and
die table speed minimizing tablet weight variation. The
appropriate paddle speed can be determined by using a
Low-Level Product Sensor force-control system that displays the standard deviation
of the compression force. The optimal feeder speed is
Variable determined by adjusting the feeder speed to achieve the
Feeder Speed lowest standard deviation in the compression force,
Adjustment which corresponds to the least weight variation.
A rectangular paddle design is typically used to
Material Hopper minimize powder mixing in the feeder. However, for
materials with poor flow characteristics (bridging in
the hopper) due to interparticle friction, a round (or
Mechanical wedge) paddle design can improve flow by forcing
Gear Drive interparticle slippage. Under these circumstances,
Train round paddles provide a mixing effect with possible
Chamber #1
Chamber #2 Chamber #3 impact on uniformity, compressibility, and dissolution.
Die Table The feeder height above the die table surface is very
Die Cavity
important to minimize product loss and prevent
scaling of low melting materials. The feeder height
Fig. 6 Material feeding system. is usually maintained between 0.05 and 0.10 mm
Tablet Compression: Machine Theory, Design, and Process Troubleshooting 3619

(0.002–0.004 in.). Very fine particles may require a problems exist, the longest time for compression should
feeder height of 0.025 mm (0.001 in.). be allowed by running at low press speeds and running
on machines with a small pitch-circle diameter.
Excess-Material Stripper. The excess-material stripper As mentioned previously, for certain types of pro-
is located immediately after the feeding system and ducts, precompression at a force level higher than that
scrapes off the excess material on the die table after of main compression may increase tablet hardness. The
weight adjustment. It is often overlooked during setup author has found that for materials that primarily
although it is one of the most critical components of undergo brittle fracture, application of a precompression
the tablet press. A brass stripper is employed, which force higher than the main compression force can result
sits flush on the die table under spring tension. The in a higher tablet hardness. However, this is typically not
material is scraped off just before the lowering cam. the case for materials with elastic properties (e.g., pro-
The brass stripper directs the excess material into the ducts prone to capping and lamination) because these
recirculation channel. A tail-over-die covers the die products require gradual application of force to mini-
cavity after scrape-off to the point of upper punch mize elastic recovery and allow stress relaxation.

Tablet–Tablet
entry. This design minimizes uncontrolled material loss Most heavy tonnage machines (80–100 kN capa-
due to flinging of material out of the die cavity at high bility for precompression and main compression) have
rotational speeds. no mechanical linkage between the upper and lower
compression rollers. Therefore, for these machines,
Precompression and Main Compression Rollers. After movement of the upper punch insertion depth does
die fill and scrape-off, the punches rotate to the pre- not result in an equal movement of the lower com-
compression station where an initial force is applied pression roller position. The compression rollers are
to the compact. The tablet is frequently partially typically mounted to a block assembly that is adjusted
formed during the precompression stage. Sub- by an eccentric or a vertical slide adjustment (Fig. 5).
sequently, the upper and lower punches move together An eccentric adjustment typically results in a slight
under the main compression rollers where the final tab- off-center alignment as the roller moves through all
let is formed. The main compression roller is usually of its possible positions, whereas a vertical slide adjust-
larger than the precompression roller. However, latest ment always maintains the roller along the center line.
advances suggest that similar sizes for precompression On older rotary tablet presses the rollers are attached
and main compression rollers with the ability to apply to a rocker arm with one side fixed to the machine roof
similar loads may result in optimal tablet formation. or base. On the upper compression roller, the other
The compression rollers are made of premium tool side of the rocker arm is mechanically linked to the
steels and are surface hardened. lower roller rocker arm. In this way, adjustment to
Because the compression characteristics of powders insertion depth results in simultaneous adjustment of
are time-dependent (the exact extent of this depen- both the upper and lower roller positions. As with
dency depends on the primary modes of deformation), the eccentric roller adjustment, rocker arm position
the final tablet properties depend not only on adjustment results in a slight off-center alignment as
maximum compression forces but also on the rate at the roller moves through all of its possible positions.
which these forces (rate of deformation) are applied Although most rotary tablet presses operate by
and removed. On a rotary tablet press, the rate of maintaining fixed roller positions during compression,
deformation is determined by the tangential velocity some designs incorporate a compression compensator
of the punch and the compression roller diameters. system in which the counterforce for compression is
The tangential velocity of the punch is a product of air pressure. This system compresses to a constant
the press speed and the die table circumference (i.e., force and allows roller movement when the preset force
die table rpm  3.14  pitch circle diameter). As is achieved. Under these conditions, potential exists to
the tangential velocity increases, the rates of com- increase the time that the force is maintained near
pression and decompression increase while the overall its peak value (approximately 90% of maximum).
compression time decreases. The roller diameter affects Compression to a constant force should theoretically
both the rate of compression and decompression. As provide a more uniform tablet hardness and more
the diameter increases, the rates of compression and uniform dissolution profiles while allowing a greater
decompression decrease. variation in tablet thickness.
Optimal compression efficiency is achieved on
machines that offer multistage compression with high Tablet Stripper. The tablet stripper scrapes off the
precompression and main compression force flexibility tablets from the lower punch and directs them down
(typically 100 kN maximum). The roller diameters should the discharge chute. On high-speed machines, special
be as large as possible to provide the lowest possible attention must be paid to the tablet takeoff to prevent
rates of compression and decompression. If compression tablet backup; modifications are necessary for shaped
 3620 Tablet Compression: Machine Theory, Design, and Process Troubleshooting

tablets, as shown in Fig. 2. On high-speed machines it and redirecting the material from the recirculation
is critical to move the tablets off the die table as channel to incorporate it into the bulk of the material
quickly as possible. Under some circumstances, reposi- in the feeder, thus preventing it from contacting the
tioning of the Plexiglas cover on the tablet stripper to lower punch directly.
provide minimal clearance between the tablet and the
cover may prevent shingling of tablets. The height of Dust Extraction. Adequate dust extraction is neces-
the lower punch at the point of scrape-off should always sary to maintain high-speed operation for extended
be checked to verify that it is not below the die table periods of time. The entire compression area should
surface. Typically the lower punch should protrude be shrouded to minimize dust infiltration into other
approximately 1–2 mm from the die table surface at press areas. Effective dust extraction minimizes dust
the point of scrape-off. For deep concave tablets, a pro- and oil contamination on the surface of the tablets,
trusion height above 2 mm may be necessary. which could produce black specs. Insufficient dust
Tablet presses equipped with single-tablet rejection extraction results in excessive material build-up on
capabilities reject tablets at the point of scrape-off. the lower and upper punches leading to tight punches.
Tablet–Tablet

Based on the compression force of the punch station, However, the proper balance of dust extraction with-
single tablets are sorted by using a compressed-air out high levels of material loss must be determined.
blow off or a mechanical fast gate. On single-sided If the dust extraction level is too high material could
machines (36 stations) both systems work well for both be extracted from the die cavities and the recirculation
large and small tablets. However, double-sided, high- channel. Furthermore, the dust extraction systems pre-
speed machines may present difficulties for large ferentially removes the fine particles. Therefore, if the
tablets at high press speeds. granulation is a direct-compression blend where the
active constituent is of fine particle size, minimum dust
Material Recirculation. Material is recirculated from extraction levels combined with minimal recirculation
the center of the turret into the feed frame. Some press may be necessary to prevent a loss of active constitu-
designs include recessed recirculation channels to mini- ents (resulting in possible low assay).
mize particle attrition and prevent excess material loss
to the vacuum system. It is critical not to recirculate Lower cam section
too much material because this can result in low pro-
duct yields and can have a detrimental effect on the The lower cam section is completely sealed from the
powder’s physical properties, which could result in compression section. It houses the lower compression
poor compressibility, uniformity, and final properties rollers, the entire lower cam track that guides the lower
(e.g., reduced dissolution rate). punches as the turret rotates, and all adjustments for the
The point of re-entry of the granulation into the fee- lower precompression and main compression roller
der corresponds to the location where the lower punch positions. Additionally, any motors necessary for auto-
enters into the fill cam when it is flush with the die table matic machine adjustment are contained in this section.
surface. Therefore, the material from the recirculation
channel is typically the material in contact with the Fill Cam. The fill cam is designed to lower the punch
lower punch face and is the first material to be filled to overfill the die cavity. Lower-punch fill cams are
for each die cavity. The effect of the material in the typically available in a variety of sizes that are changed
recirculation channel should always be evaluated when depending on the final fill depth as determined by the
compression problems occur, which can be associated weight regulation cam. Press manufacturers rec-
with the lower punch. An example would be a ommend a fill cam in which the weight regulation
case where a granulation exhibits picking only on the cam operates in the approximate center of the fill
lower punch and not routinely on all tablet presses. cam. Typical fill cams have a range of approximately
Additionally, this granulation is slightly underlubri- 10 mm with an increment range of 4 mm (e.g., 0–10,
cated, very friable, and undergoes minimal brittle frac- 4–14, 8–18, and 12–22 mm). Special-order or very shal-
ture. Previous experiments showed that, as particle size low fill cams are also available (e.g., 0–6 mm). Alterna-
decreases, ejection force and picking tendencies tively, some manufacturers offer a greater selection of
increase. It was determined that, for this product, cams with a narrower range and lower increment
excessive material recirculation resulted in a reduction (e.g., 5.5 mm range with an increment of 2 mm). These
in particle size and an increase in picking tendencies. cams should offer greater precision in fill in certain
However, the picking problem was seen only on the cases. However, with the narrower range, minor changes
lower punches because the reduced particle size in the granulation density could result in the necessity to
material came into direct contact only with the lower change fill cams throughout the course of the run.
punches. The problem was minimized by reducing Flexibility in fill cam options offers advantages for
the amount of material in the recirculation channel specific problem areas. For example, if a material is
Tablet Compression: Machine Theory, Design, and Process Troubleshooting 3621

very abrasive, a shallow fill cam should be chosen to The lower dosing rail and holding-down cam should
minimize the amount of material that is removed from always be made of a tough, abrasion-resistant material;
the feeder and subsequently returned to the feeder and aluminum–bronze alloy is highly recommended. The
the recirculation channel. A deep fill cam can cause condition of the dosing rail and the holding-down
overpacking of the feeder, which could result in jam- cam should be checked every 6 months to ensure opti-
ming, temperature increases, and overmixing of granu- mal performance. If significant wear is observed, the
lation or lubricant. cams should be replaced or, in the case of the holding-
down cam, reworked to provide a tight fit.
As the lower punch leaves the dosing unit, it is pulled
Weight Regulation Cam. The lower punch travels
down slightly (approximately 2 mm) by the lowering
from the fill cam to the weight regulation cam, which
cam. Periodically the condition of the lowering cam
determines the final volume of material that remains
should be checked to ensure proper lowering of the
in the die cavity after scrape-off. Proper design and
punch in order to minimize weight variation.
operation of this unit is essential to ensure uniform
Many modern presses are equipped to measure the

Tablet–Tablet
tablet weights. In general, the unit should operate in
tightness of the lower punches. This function is critical
a manner to ensure smooth punch travel minimizing
to minimize machine damage as well as cam wear. As
punch chatter as the lower punch is raised to a precise
mentioned above, excessive cam wear increases tablet
and constant height.
weight variation. Lower-punch tight sensors are typi-
The weight-regulation unit consists of several criti-
cally mounted in either of the two locations. The tran-
cal components that determine its efficiency of oper-
sition cam from the fill cam to the weight regulation
ation as shown in Fig. 7. The lower punch rides on
unit is an ideal location to measure lower punch tight
the dosing rail that maintains the lower punch at a con-
forces because it is designed to raise the level of the
stant height at the final fill depth. In order to minimize
lower punch to the final dosing height. In addition,
vertical movement (and subsequently punch chatter)
the lowering cam can measure the counterforce to pull-
the head of the lower punch is held tight against the
ing down the lower punch.
lower dosing rail by the holding-down cam which is
spring loaded. To ensure that the holding-down cam
Lower Punch Brakes. Most rotary tablet presses are
is tight against the head of the lower punch, a 1–5 mm
equipped with lower-punch brakes that are Teflon
(0.04–0.20 in.) gap should be maintained between this
tipped and spring loaded to apply constant pressure
cam and the lower dosing cam when the lower punch
to the lower punches. Alternatively, some manufac-
rests on the dosing cam. During press setup, the fit
turers apply pressure to a friction belt that provides
can be easily checked by placing a lower punch in the
resistance on the lower punches. The lower-punch
weight regulation unit and verifying the absence of
brakes act as a ‘‘retention’’ system for holding the
vertical movement of the punch. This function is critical
lower punches in place during press setup. More
to minimize tablet weight variation at high speeds. On
importantly, these systems help to minimize lower-
many press designs, the weight regulation unit contains
punch chatter at high press speeds thus minimizing
a safety cam on its inside.
tablet weight variation. Some press manufacturers
use the lower punch seals to retain the lower punches.

Precompression and Main Compression Rails. The

precompression rail provides the transition support
for the lower punch from the weight regulation unit
to the precompresison roller, while the main com-
pression rail provides the transition support from the
precompression roller to the main compression roller.
Safety Holding The optimal press designs provide positive support with
Rail Down Cam these cams by ensuring that the lower punch head flat is
always in contact with the rail surface. In this way, there
Lower Punch is no abrupt vertical movement of the lower punch as it
Gapping passes to the compression rollers. Vertical movement of
(1–5mm)
the lower punch before precompression and between
the compression stations can cause the introduction
Lower Dosing Cam of air into the bottom of the compact, resulting in cap-
ping at the lower-punch face. Many presses rely on the
lower punch brakes or seals to prevent this type of
Fig. 7 Weight regulation unit. movement. Under these conditions, vertical movement
 3622 Tablet Compression: Machine Theory, Design, and Process Troubleshooting

of the lower punches will gradually occur as these com- Force Overload System. Most tablet presses are
ponents wear causing periodic compression problems. equipped with force overload systems designed to pre-
vent machine and punch damage. As stated previously,
Adjustment of Lower Precompression and Main Com- the compression force is not a set value, rather a
pression Roller Thickness. The position of the lower measured value obtained from the fixed punch distance
compression rollers is adjusted from the lower cam sec- and the quantity of material in the die. On most tablet
tion. As shown in Fig. 8, the position of the lower roll- presses, a maximum allowable compression force can
ers relative to the upper rollers (i.e., insertion depth) be set. This force setting is actually the counterforce
determines the tablet edge thickness. Typically, the to the measured compression force. If the compression
machine-control panel shows edge thickness on the force exceeds this counterforce, the compression
indicator. However, adjustment of the edge thickness assembly will back-off thus reducing the force. Most
actually results in adjustment of the lower roller pos- tablet presses use a hydraulic, air, or spring-loaded sys-
ition only. For machines that have no mechanical link tem on the lower compression assemblies (both pre-
between the upper and lower compression rollers, the compression and main compression). In these systems
Tablet–Tablet

tablet edge thickness indication on the control panel the hydraulic or spring systems are calibrated to the
is only valid at the specific insertion depth that was measured force in the die and move instantaneously
set during edge thickness calibration. However, for during an overload condition. Some of the more recent
some of the electronic, fully automated machines, the rotary tablet presses use strain gauges to measure
machine automatically moves the lower compression the actual compression force for force overload
roller during the insertion depth adjustment to main- (as opposed to force control). In these systems, the
tain the same tablet edge thickness. machine mechanically moves the compression assembly
when the measured force is exceeded. Therefore, since
Ejection Rail. After compression the lower punch these systems do not react as quickly as hydraulic, air,
impacts the ejection rail (or on some machines an ejection or spring systems they typically include a safety margin
roller). Upon impact the tablet is broken free from the die to initiate an overload condition prior to reaching the
side wall and begins to move up the die as the machine actual set value (e.g., 95% of maximum entered value).
rotates. The ejection rail should be made of a tough, abra-
sion resistant material such as aluminum bronze alloy. Lower mechanical section

Scrape-Off Rail. After riding up the ejection rail, the The lower mechanical section houses the main drive
lower punch rides on the scrape-off rail to provide a motor, the gearbox, the hydraulic pump, the lubri-
constant height for tablet scrape-off. The height of the cation pump, and the signal wire distribution. Proper
lower punch scrape-off can be adjusted to optimize the sin- venting and cooling of the lower mechanical section
gle tablet rejection height or the tablet scrape-off height. is essential to prevent machine damage and minimize
heat generation. This section should be equipped with
a cooling system for products that are sensitive to heat
generation (e.g., contain low melting point components
UTLwi that are prone to picking and sticking).

UTL w- Electrical section

UCL w- The electrical section contains all electrical controls

Weight (mg)

and components (e.g., programmable controllers,

Target F- relays, contacts, and fuses), the signal wire distribution,
and the integrated or remote force control systems. On
LCL w- many machines the electrical section is connected to
the front of the press, thus minimizing space require-
LTL w- ments. However, modern fully automated machines
use computers that may be sensitive to machine
LTLwi vibration, dust, and heat. Therefore, these electronics
should be located remotely from the machine.
LTLFi LTL F- LCL F- UCL F- UTL F- UTLFi
Target F- Lubrication system
Force (KN)
Most high speed rotary tablet presses employ automatic
Fig. 8 Force versus weight relationship. lubrication systems during operation. Effective punch
Tablet Compression: Machine Theory, Design, and Process Troubleshooting 3623

lubrication is essential for the movement of the punches and the lower punches were observed to be warm. They
in the turret, in the cam tracks, and under the com- were removed, cleaned, lubricated, and placed back in
pression rollers. Punch-lubrication systems allow high the machine. Upon resuming production, the tablets
speed operation over extended periods of time while returned to their original characteristics. In this case,
minimizing cam and tool wear and reducing heat the lack of a punch lubrication system combined with
generation. batch campaigning (extended running conditions)
The lubrication pump is typically maintained in the resulted in a temperature increase in the machine.
lower mechanical section and allows variation of both For this elastic material, the increased temperature
the lubrication interval and time duration. The quan- resulted in greater plasticity and more stress relaxation,
tity of oil delivered is normally determined by the oil improving the compressibility of the product.
distribution nozzles connected to the distribution Although under many circumstances this effect would
manifolds. The upper punches are usually lubricated be beneficial, in the present case the improved com-
on the punch head flat via a felt pad located in the pressibility resulted in out-of-specification product.
upper punch dwell cam or the upper punch lowering The problem was resolved by removing the punches

Tablet–Tablet
cam. On many modern designs the upper punch barrel after each batch for cleaning and lubrication.
is also lubricated. The punch barrel is lubricated via
radially drilled holes in the turret that transports oil
to the punches by rotational forces, or by overlubricat- Instrumentation
ing the punch neck and allowing gravity to transfer the
oil down the barrel. Because the upper punch shaft has Modern rotary production tablet presses are typically
the greatest area of contact with the upper turret, lubri- equipped to measure precompression and main com-
cation of the punch shaft is critical to allow high-speed pression forces. Additionally, measurement and moni-
operation. Poor lubrication of this area can result in toring of tablet ejection force can prove to be beneficial
heat generation and metal expansion, ultimately caus- for specific problem products and for production
ing machine seizure and severe damage. troubleshooting. However, for most pharmaceutical
The lower punches are lubricated at the punch neck. products proper product development and optimization
Oil distribution lines are frequently provided to lubri- work eliminate the need to instrument a production
cate both sides of the punch neck. Subsequently, gravi- machine for ejection force. Rotary tablet presses can
tational forces distribute the oil over the head of the also be equipped to measure both upper punch and
punch. The lower punch barrel is typically lubricated lower punch pull-down forces. These measurements
by radially drilled holes in the turret. are primarily made to detect tight-running punches
All presses equipped with punch-lubrication systems and are necessary on production machines only if the
require oil and dust seals to prevent oil contamination machine monitoring system uses direct force measure-
in the product and dust contamination in the turret ment for these functions. Tablet scrape-off force can
punch sockets. These seals are normally double lipped, also be measured, but this is only recommended on
designed to strip oil on one side and powder on the other. development machines. Scrape-off forces are typically
As mentioned previously, some press designs use the lower below 6 N. Therefore, instrumentation of a tablet strip-
punch seals to retain the lower punches. per requires highly sensitive instrumentation that is
Inadequate punch lubrication can lead to excessive easily damaged.
heat generation, which could affect tablet properties. Precompression and main compression forces are
An example is a granulation that primarily undergoes normally measured for the upper punches.[20,21] These
elastic and plastic deformation. This product was nor- forces are typically measured using strain gauges
mally run on a tablet press without an automatic lubri- arranged in a full wheatstone bridge.[22] Strain gauges
cation system. Production requirements resulted in are basically resistors applied to the metal surface in
batch campaigning. At the beginning of the production a specific orientation. Under load, the member deflects
campaign, the punches were lubricated and placed into and the strain changes the resistance of the gauge. The
the machine. Over the course of the first batch, the pro- change in resistance is proportional to the applied force.
duct was easily maintained within tablet hardness and However, due to design differences, some machines
thickness specifications. However, as the campaign measure the lower compression forces as opposed to
transitioned into batches two and three, tablet hard- upper compression forces. The compression force should
ness tended to increase while the thickness remained be measured as close to the compression event as pos-
constant. Machine adjustments were made to maintain sible. For the most accurate and reproducible readings,
the hardness and thickness within specification, but in the strain gauge should be in line with the compression
the end the hardness could not be maintained below event as opposed to off center at a remote location. This
the maximum limit without exceeding the thickness is easily accomplished on most rotary tablet presses by
specification. At this point the machine was inspected using a shear pin to support the compression rollers.
 3624 Tablet Compression: Machine Theory, Design, and Process Troubleshooting

However, the pin must not be rotated when the position hardness. If the tablet weight is outside of the estab-
of the compression rollers is changed. Therefore, this lished control limit, the operator increases or decreases
system is ideally suited for machines that change roller the weight by adjusting the fill depth (increasing fill
position by a vertical slide mechanism as opposed to an depth increases tablet weight and decreasing fill depth
eccentric mechanism. Shear pins are typically custom reduces tablet weight). If either the tablet hardness or
manufactured, where quality depends not only on the thickness requires adjustment, the operator typically
pin design but also on the strain gauge receptivity and adjusts the tablet edge thickness on either main com-
arrangement. pression or precompression. Since there is normally
Many modern rotary tablet presses use off-the-shelf an inverse relationship between the tablet hardness
load cells for force measurement. These load cells are and thickness, the operator usually reduces edge thick-
highly accurate, durable, and easily replaced and cali- ness to increase tablet hardness (or decrease thickness)
brated. However, the final accuracy and repeatability or increases edge thickness to decrease tablet hardness
of force measurement in the machine not only depend (or increase thickness).
on the quality of the load cell, but also on the design of Force and weight control systems use the same basic
Tablet–Tablet

the compression assembly and the placement of the concepts as conventional machines. Force control sys-
load cell within the assembly. tems monitor the tablet compression force and adjust
Machines that utilize rocker arms with a mechanical the fill depth to maintain a constant force. Force con-
linkage between the upper and lower compression assem- trol systems alone compensate for flow and density
blies are normally instrumented by applying a strain variations in the granulation, providing a constant fill
gauge to the upper rocker arm or to the mechanical link- quantity. Weight control systems, on the other hand,
age connecting the assemblies. The point of strain gauge work in conjunction with the force control system as
application is ‘‘necked-down’’ to increase the sensitivity a secondary control loop and replace the manual func-
of the member. These instrumented members should be tion of the machine operator, periodically removing tab-
calibrated in the machine to account for the effect of let samples to test tablet weight, thickness, and hardness.
other machine members on the measured force. If the weight control system indicates that the tablet
The force measurement system (strain gauges or weight must be adjusted, the force control setpoint or
load cells) should be calibrated on a yearly basis or the tablet edge thickness is altered resulting in a change
after the compression assembly has been disassembled in the fill depth from the force control system.
for any reason. The calibration should be made in the
machine to assure accuracy. It is typically performed
using a modified punch assembly and a calibrated load Force Control
cell that are rotated under the compression roller to
produce a load. The output of the machine force During tablet compression, the distance between the
measurement system is compared to the output of rollers remains constant unless a machine adjustment
the calibrated load cell. Many machine manufacturers is made to change tablet hardness or thickness.
use this single-point calibration to modify the strain Additionally, all tooling dimensions (tooling length
gauge factor so that the two outputs are the same at and die cavity size) are constant within established
this load level. Subsequently, different load levels are standards. Under these conditions, for a specific
tested and the error between the machine force material of uniform density, if the same volume of
measurement system and the calibrated load cell is material is delivered to each die, the maximum mea-
determined. Alternatively, loads can be applied to the sured compression force for each punch station is the
system ranging from the minimum to maximum com- same. If, on the other hand, different volumes of
pression forces. The outputs from the calibrated load material are delivered to each die, the maximum
cell and the machine force measurement system are measured compression force for each punch station is
recorded and a linear regression is performed on the different. On this basis, adjustment of fill depth (fill
data to calculate a new strain gauge factor across the volume) to maintain a constant compression force
entire force measurement range. Subsequently, differ- should result in a constant tablet weight. This concept
ent load levels are tested and the error between the is the general basis of all rotary tablet press force
readings is calculated. control systems.
The force control systems assume a linear relation-
ship between tablet weight and compression force for
PRESS CONTROL AND AUTOMATION a particular granulation, tooling set, and machine tab-
let edge thickness (i.e., distance between compression
Conventional rotary tablet presses are controlled by rollers). By establishing the relationship between com-
periodically taking tablet samples from the discharge pression force and tablet weight at a specific machine
chute and checking their tablet weight, thickness, and tablet edge thickness (as shown in Fig. 8), a force
Tablet Compression: Machine Theory, Design, and Process Troubleshooting 3625

control system is able to maintain a constant tablet the set point of 10 kN (outside of the tolerance set by
weight by maintaining a constant compression force. the manufacturer) then the force control system adjusts
Additionally, a force control system is capable of the fill depth to return the force to 10 kN. If the com-
monitoring every compression force and rejecting pression force is outside the average tolerance limits
tablets when the forces exceed specific established lim- (below 7 kN or above 13 kN), the machine will shut
its (for both average forces and individual forces), down and reject the tablets. This condition should cor-
thereby essentially monitoring every tablet weight. respond to average tablet weights outside of the
The first step in using a force control system is to 97.0–103.0 mg limits. Typically, the force control sys-
establish the force versus weight relationship to allow tem is capable of adjusting the fill depth to maintain
calculation of the appropriate force control set points the compression force well within the tolerance limits.
which correspond to the desired weight control points. However, if the material has poor flow characteristics
Table 1 gives example of weight control points and tol- and exhibits bridging followed by surging, the system
erance limits for a theoretical product. may be unable to compensate quickly enough to pre-
The tablet press is initially run at target conditions vent these out of control conditions.

Tablet–Tablet
to make the product within specifications. For During the entire operation, if any of the individual
example, the tablets are made at target conditions of measured compression forces goes outside of the indi-
100 mg at an average compression force of 10 kN. vidual tolerance limits of 5 kN or 15 kN (correspond-
After establishing this point, the fill depth is adjusted ing to individual tablet weights below 95 mg or above
to either increase or decrease the tablet weight and 105 mg), these individual tablets are rejected at the
the corresponding compression force is measured. In point of tablet scrape-off. Some machine designs are
the present example, the tablet weight was increased not effective at sorting individual tablets reliably and
to 103 mg and the average measured compression force reject multiple tablets if this condition occurs. For
was approximately 13 kN. This procedure should be most machine designs, the user can specify a maximum
repeated for several different tablet weights. The data number of individual tablets that can be sorted per
are used for regression analysis to calculate the punch location or per batch before the machine shuts
required force set points that correspond to the weight down. Exceeding these maximum limits may indicate
control points as shown in Fig. 8 and Table 2. a problem punch or excessive weight variation for the
As an alternative to performing regression analysis, batch, possibly related to a setup problem.
fill depth can be adjusted to achieve each average As described, a force control system maintaining a
weight requirement and the resultant compression constant compression force will adequately compen-
force can be recorded and set. sate for variations in granulation density, thus provid-
During normal production with the force control ing more uniform tablet weights. However, operation
system in operation as specified above, the tablet press of this system still requires an operator to periodically
will operate to maintain the constant compression check tablet weight, thickness, and hardness. If during
force of 10 kN by adjusting the fill depth (Fig. 9). Most a weight check the operator determines that the aver-
force control systems do not require the user to input age tablet weight has gone beyond the control limit,
the upper and lower force control limits for average while the average compression force is still being main-
compression force that are typically set by the manu- tained at its set point, the operator must take one of
facturer within tighter tolerances than those demanded the following actions:
by the weight requirements. The control system typi-
cally calculates the average compression force every  The system is placed in manual mode (shut-off
revolution and compares it to the force set point. If force control system) and the tablet weight adjusted
the average measured compression force varies from back to target by increasing or decreasing the fill

Table 1 Tablet weight-control points and tolerance limits

Item Description Specification (mg)
UTLWi Upper tolerance limit of individual tablet weight 105.0
UTLW Upper tolerance limit of average tablet weight 103.0
UCLW Upper control limit of average tablet weight 101.5
TargetW Target of average tablet weight 100.0
LCLW Lower control limit of average tablet weight 98.5
LTLW Lower tolerance limit of average tablet weight 97.0
LTLWi Lower tolerance limit of individual tablet weight 95.0
 3626 Tablet Compression: Machine Theory, Design, and Process Troubleshooting

Table 2 Force setpoints

Item Description Specification (kN)
UTLFi Upper tolerance limit for individual compression force 15.0
UTLF Upper tolerance limit for average compression force 13.0
UCLF Upper control limit for average compression force 11.5
TargetF Target for average compression force 10.0
LCLF Lower control limit for average compression force 8.5
LTLF Lower tolerance limit for average compression force 7.0
LTLFi Lower tolerance limit for individual compression force 5.0

depth as necessary. Once the weight is within of the granulation or to changes in the machine over
Tablet–Tablet

requirements, the force control set point is changed the course of the run (such as temperature changes).
(and all other limits by the same amount) to the Tablet presses that use a compression compensator
current value that is displayed and the machine is system, which in turn compress to constant com-
returned to the automatic mode. pression forces and allow roller movement, operate
 While in the automatic mode, the force control set by the same control theories as those presented for
point is increased or decreased to increase or decrease force control. However, these systems measure roller
the tablet weight, respectively. For example, by displacement as opposed to compression force and
increasing the force set point, the control system relate it to tablet weight.
increases fill depth to achieve the higher compression
force requirement thus increasing tablet weight. The Weight Control
machine is allowed to stabilize and tablet weight is
checked. Adjustments are made until tablet weight Weight control, as a secondary control loop to force
is within specifications. control, allows automation of the tableting operation.
 While in the automatic mode, the machine tablet The weight control system (Fig. 10) maintains the same
edge thickness is increased or decreased to increase limits as those presented for force control (Fig. 9). The
or decrease the tablet weight, respectively. For machine assumes the place of the operator and period-
example, by increasing the machine tablet edge ically samples the tablet press to determine the tablet
thickness, the measured compression force is weight. The sampling requirements should be set to
decreased. The force control system will then the same interval and number of tablets as those
increase fill depth to return the compression force required for manual operation. Most weight feedback
to its previous value thus increasing tablet weight. systems measure individual tablet weights and calcu-
late the average weight for feed back purposes.
Minor changes in the force to weight relationship In the example given above, it is assumed that the
over the course of a compression run are common. weight feedback system will measure 10 individual
These may be due to changes in the compressibility tablet weights every 15 min. The force control system
is in operation and is maintaining the average com-
pression force at 10 kN. If during each 15 min. check,
the average weight of 10 tablets is within the average
Individual Reject Region
UTL Fi = 15.0
Shutdown Region
UTL F- = 13.0 Individual Reject Region
Adjustment Region1 UTLwi = 105.0
Force (KN)

UCL F- = 11.5 Shutdown Region

Time UTLw = 103.0
Target F- = 10.5 Adjustment Region
UCLw = 101.5
Force (KN)

LCL F- = 8.5
Adjustment Region1 Targetw = 100.0 Time
LTL F- = 7.0
Shutdown Region LCLw = 98.5
LTL Fi= 5.0 Adjustment Region
Individual Reject Region LTLw = 97.0
Shutdown Region
Note 1: Adjustment region corresponds to weight control system. Force adjustment LTLwi = 95.0
typically occurs within a very narrow band width as shown in the shaded region. Individual Reject Region

Fig. 9 Force control system. Fig. 10 Weight control system.

Tablet Compression: Machine Theory, Design, and Process Troubleshooting 3627

control limits (Fig. 10), no machine adjustments from shut-off the machine if tolerance limits are exceeded.
the weight control system will be made. Alternatively, In this way, fully automated operation is possible with-
if the average tablet weight goes beyond the average out operator testing.
control limits (above 101.5 mg or below 98.5 mg) the
weight control system will initiate a change in one of
TROUBLESHOOTING
the following ways depending on the machine:
A proper understanding of a material’s compression
Increase or decrease the force control setpoint to
characteristics combined with a knowledge of tablet
increase or decrease the tablet weight, respectively.
compression equipment allows efficient troubleshoot-
The new force control set point is calculated by
ing of production problems. Although there is no sub-
means of the machine manufacturer’s control
stitute for a robust granulation, a product can be
algorithm. Typically after adjustment to the new
optimized by examining all of the different machine
force control set point, the weight control system
factors that can affect performance. By applying a vari-
should resample and verify that the average tablet

Tablet–Tablet
ety of the concepts discussed here, a large variety of
weight is now within the average control limits.
processing problems can be eliminated.
Increase or decrease the machine tablet edge thick-
ness to increase or decrease the tablet weight,
Tablet Weight Variation
respectively. The new machine tablet edge thick-
ness setting is calculated by the manufacturer’s
Excessive tablet weight variation can be caused by a
algorithm. After the new setting has been made
variety of factors. For many granulations, the inherent
and the fill depth adjusted to maintain constant
poor flow characteristics of the material may be the rate
compression force, the weight control system
limiting step, and simply slowing down the machine may
should resample and verify that the average tablet
reduce weight variation. Additionally, optimization of
weight is now within the average control limits.
the feeder paddle speeds to minimize the standard devi-
ation in the compression force should help to minimize
The weight control systems that change the force
weight variation. If weight variation is excessive, the
control setpoint result in a change of the compression
following machine components should be examined:
force throughout the run with a relatively constant tab-
let thickness. On the other hand, by changing the
 The tightness of the hold-down cam should be
machine tablet edge thickness, the compression force
examined to verify that it is not excessively worn
remains relatively constant throughout the run while
and is holding the lower punch tight against the
the tablet thickness varies. Since these adjustments
dosing cam. If the previous product resulted in tight
are usually relatively small, both methods of machine
lower punches, premature wear may have occurred
adjustment typically produce similar results.
on this cam, causing increased weight variation.
If during any check the average tablet weight
 The condition of the lower punch pull-down cam
exceeds the average tolerance limits (below 97 mg and
should be examined to verify that it is not overly
above 103 mg) or an individual tablet exceeds the indi-
worn and that it drops the lower punch to pull the
vidual tolerance limits (below 95 mg and above
material in the die cavity below the die table surface.
105 mg), the machine will shut off and reject the
 Both the condition and position of the excess
tablets. It is important to note that individual tablet
material stripper should be examined to assure that
weights exceeding the tolerance limits indicate that
it sits snug and level on the die table surface for
the tablet rejection system is not functioning properly.
complete scrape-off.
 Different types of feeder paddles can be used to
Control and Monitoring of promote flow (e.g., round feeder paddles are used
Weight/Thickness/Hardness for materials that exhibit bridging). The feeder
speed should be optimized to minimize force and
Tablet presses equipped to measure tablet weight, weight variation.
thickness, and hardness use the same concepts for force  The best fill cam size is that where the fill depth is
and weight control as presented here. These systems centered in its range.
offer the additional flexibility of testing and controlling  A minimum amount of material recirculation is
both tablet thickness and hardness. However, most necessary to provide steady flow and fill. Too much
machine manufacturers and tablet manufacturers have recirculation can result in material back-up and
found that additional control of either tablet thickness reduction in the granulation particle size. The recir-
or hardness is both difficult and costly. It is cost effec- culation channel must be free of obstructions (i.e.,
tive to only monitor tablet thickness or hardness and broken tablets).
 3628 Tablet Compression: Machine Theory, Design, and Process Troubleshooting

 If large tablets are being produced requiring deep capping problems, both precompression and main
fill depths, then insertion depth should be increased. compression forces should be maximized.
Otherwise, as the punches pass from a deep fill to a  Press speed is reduced in order to increase total
shallow insertion depth, the uncompressed granu- compression time.
lation will be pushed out of the die cavity resulting  Formulations sensitive to lubricant levels may exhi-
in material loss. bit low hardness due to overmixing in the feeder. If
the formulation contains a significant quantity of
magnesium stearate or there is a shift in particle size
Product Yield thus extending the lubricant differently, overmixing
can reduce tablet hardness. In this case, the feeder
Product yield can be affected by a number of factors. speed should be reduced to a minimum.
Unfortunately, yield problems are not noticed until
after the loss occurs. However, by paying attention to
Capping and Lamination
the following areas during setup, these problems can
Tablet–Tablet

be minimized:
Tablet capping and lamination typically create the
most difficult problems, due to a variety of causes.
 Excessive material loss can be avoided by ensuring
Identification of the cause often leads to the solution.
that the excess material stripper is flush against the
The basic concepts to alleviate these problems center
die table. Otherwise the material will be sucked into
on minimizing elastic behavior while promoting plastic
the dust extraction system.
deformation. Depending on the exact nature of the
 The feed frame height should be maintained
problem, this can be achieved from a formulation per-
between 0.05 and 0.10 mm (0.003–0.004 in.). For very
spective by modifying the formula to incorporate a
fine particle size granulation, the clearance should be
plastically deforming matrix, by adding components
reduced to 0.025–0.05 mm (0.001–0.002 in.).
to enhance bonding, or by increasing the moisture
 The fill cam size should be reduced to minimize
level. Alternatively, from a machine perspective the
overfilling and material recirculation.
following guidelines should be followed:
 Material in the material recirculation channel
should be maintained at a minimal level. As more
 The rate of force application should be reduced by
material is recirculated, the likelihood for loss is
applying the compression force as gradually as
greater. The piece guiding the material from the
possible. This can be accomplished by lowering
recirculation channel to the feeder must be properly
the press speed or using a machine with a small
positioned.
pitch circle diameter.
 If the insertion depth is too shallow relative to the
 The ratio of precompression to main compression
fill depth, material will be pushed out of the die
should be modified with gradual application of
and lost to the dust extraction system.
the precompression force followed by main com-
 Excessive feeder speeds cause excess material recir-
pression. A precompression force that is too high
culation and increase material loss.
can be harmful.
 It may be necessary to reduce press speed. As the
 The effect of reducing the compression force should
press speed is increased, the turret rotational forces will
be evaluated. In many circumstances, overcompres-
sling more material to the outside of the turret from
sion of a granulation will result in failure.
both the recirculation channel and the die cavity.
 A machine with large compression-roller diameters
should be used to minimize the rate of force application.
 Die cavity wear must be investigated and the con-
Low Hardness
dition of the die cavities examined. If the dies are
very worn, slip-stick behavior may occur during
Tablet hardness is affected by many factors. For trou-
tablet ejection resulting in tablet failure.
bleshooting purposes, it should be determined if the
 Curled/damaged punches promote tablet capping.
low hardness is due to capping or non-compressibility.
Under these conditions the tools should be
For formulations that exhibit low hardness without
reworked or replaced.
capping, the following guidelines are helpful:

 For multi-stage compression, a machine should be Picking and Sticking

equipped for both precompression and main com-
pression with large diameter rollers. Tablet picking and sticking problems are typically
 The ratio of precompression to main compression related to formulation issues. However, in small scale
should be adjusted. For tablets that exhibit no manufacturing these problems do not occur frequently.
Tablet Compression: Machine Theory, Design, and Process Troubleshooting 3629

Regardless, once a product is approved, it is difficult to  Tablet stripper: The impact point of the stripper
make significant formulation changes. To minimize these can cause chipping. Both the height and position
problems the following areas should be considered: of the tablet stripper must be checked. Damaged
punches must be replaced or repaired.
 Heat of compression: Excessive heat generation  Air assist: If tablet jams cannot be eliminated by
during compression will increase the picking tend- modifying the tablet stripper, an air assist blow-
ency of a low melting material. Use of cooling sys- off at the tablet stripper may solve the problem.
tems for the compression section or lower  Static eliminator: In the production of lightweight
mechanical section may be helpful. tablets in a low humidity environment, static electricity
 Press speed: Lowering press speed and com- may cause tablet back-up. In this case, installation of a
pression force reduces heat generation. Lower press static eliminator will improve tablet discharge.
speeds extend the contact time between the material
and the punch face.
 Precompression force: Elimination of precom-

Tablet–Tablet
CONCLUSIONS
pression may prevent picking. For example, some
materials pick if the compression force is too low.
Optimal manufacturing of tablets requires good equip-
Therefore, application of precompression at low
ment and materials. The equipment is only a tool to
forces may result in tablet picking.
achieve the final goal of high quality tablets. However,
 Tool condition: The condition of press tooling
if the materials have marginal flow and compressibility
should always be evaluated when picking occurs.
problems, machine flexibility allows the production of
Polishing the punches and application of various
the highest quality product possible.
coatings to the tools may help to eliminate picking
for certain materials.
 Start-up conditions: Start-up should always be
close to optimum conditions to prevent fouling REFERENCES
the punch faces. If maintaining the force above a
minimum is necessary to prevent picking, starting 1. Higuchi, T.; Arnold, R.D.; Tucker, S.J.; Busse, L.W. J. Am.
Pharm. Assoc. 1952, 41, 93–96, Sci. Ed.
up near aim conditions of compression force pre- 2. Train, D. J. Pharm. Pharmacol. 1956, 8, 745–761.
vents initial picking. 3. Nystrom, C. Drug Dev. Ind. Pharm. 1993, 19, 2143–2196.
 Tablet stripper: The point of impact of the lower 4. Wray, P.E. Drug Dev. Ind. Pharm. 1992, 18, 627–658.
5. Hoblitzell, J.R.; Rhodes, C.T. Drug Dev. Ind. Pharm. 1990,
punches should be repositioned relative to the tablet 16, 469–507.
stripper. Under many circumstances the stress of impact 6. Marshall, K. Compression and consolidation of powdered
on the stripper can cause failure and sticking in logos. solids. In The Theory and Practice of Industrial Pharmacy,
3rd Ed.; Lea & Febiger: Philadelphia, 1986.
7. Parrot, E.L. Compression. In Pharm. Dosage Forms:
Tablet Jams and Chipping Tablets, 2nd Ed.; Marcel Dekker, Inc.: New York, 1990; 2.
8. Goetzel, C.G. Treatise of Powder Metallurgy; Interscience
Publishers, Inc.: New York, 1949; Vol. 1, 259–312.
Tablet jams and chipping at the stripper reduce tablet 9. Heistand, E.N. Int.: J. Pharm. 1991, 67, 217–229.
quality and may contaminate the feeder resulting in 10. Hiestand, E.N.; Smith, D.P. Int. J. Pharm. 1991, 67,
weight variation. High speed machines typically have 231–246.
11. Reier, G.E.; Shangraw, R.E. J. Pharm. Sci. 1966, 55,
greater problems than slower machines. These prob- 510–514.
lems are solved by focusing on the tablet stripper and 12. Hanus, E.J.; King, L.D. J. Pharm. Sci. 1968, 57, 677–684.
lower punch ejection height. 13. Rankell, A.S.; Higuchi, T. J. Pharm. Sci. 1968, 57 (4), 574–577.
14. Fette Technical Bulletin 8004; Fette America Inc.:
Rockaway, NJ.
 Scrape-off height: For many tablet types (e.g., 15. Esezobo, S.; Pilpel, N. J. Pharm. Pharmacol. 1986, 38, 409–413.
deep concave tablets) the height of the lower punch 16. Gunsel, W.C.; Kanig, J.L. Tablets. In The Theory and Prac-
tice of Industrial Pharmacy, 2nd Ed.; Lea & Febiger:
at the point of scrape-off must be increased to Philadephia, 1976.
ensure that the tablet is removed completely from 17. Hiestand, E.N.; Peot, C.B.; Ochs, J.E. J. Pharm. Sci. 1977,
the die when it impacts the tablet stripper. 66, 510–519.
18. Bateman, S.D.; Rubinstein, M.H.; Thacker, H.S. Pharm.
 Modified tablet stripper: For shaped tablets, a Tech. Int. June 1990, 30–36.
modified tablet stripper that removes the tablets 19. Rippie, E.G.; Morehead, W.T. J. Pharm. Sci. 1994, 83,
from the die table quickly prevents tablet back-up 708–715.
20. Watt, P.R. Tablet Machine Instrumentation in Pharma-
and breakage. ceutics: Principles and Practice; Ellis Horwood: Chichester,
 Height of plastic cover: If the tablets exhibit lay- UK, 1988.
ering or shingling, the height of the stripper cover 21. Oates, R.J.; Mitchell, A.G. J. Pharm. Pharmacol. 1994,
46 (4), 270–275.
should be lowered to just accommodate one tablet 22. Schmidt, P.C.; Vogel, P.J. Drug Dev. Ind. Pharm. 1994, 20,
between the cover and the die table. 921–934.
 Tablet Evaluation Using Near-Infrared Spectroscopy
Christopher T. Rhodes
Karen Morisseau
College of Pharmacy, University of Rhode Island,
Kingston, Rhode Island, U.S.A.

INTRODUCTION The mathematical expression relating the component

property (or properties) to absorbance is called a cali-
Near-infrared spectroscopy (NIRS) continues to grow in bration model (also referred to as an algorithm). Using
Tablet–Tablet

importance as a useful analytical technique. It offers sophisticated spectral software, the analyst can corre-
unique potential as a rapid, non-destructive method of late sample spectra to laboratory data, develop a cali-
quantitative and qualitative evaluation. NIRS has been bration model, and apply that model to similar, new
used extensively in the food and agricultural industries samples to predict constituent properties.
for many years to determine moisture, protein, and starch
content in grains.[1] The pharmaceutical industry has
been cautiously slow to accept NIRS as a commonly used THEORY
technique, probably because of the absence of primary
absorption bands. In recent years, an increasing amount The NIR region of the electromagnetic spectrum is
of academic research is being carried out on the theory from 800 to 2500 nm. The segment from 1100 to
behind NIR. The use of NIRS for pharmaceutical 2500, known as the Herschel region,[13] is the range
applications has grown owing, in part, to technological most often used in the analysis of pharmaceutical pro-
advances in instrumentation and software. ducts. In the NIR region, the radiation can penetrate
Several reviews[2–7] have been published recently, compacted materials such as tablets, providing a vast
attesting to its increasing popularity. Textbooks[8] amount of spectral information about the sample.
relating to pharmaceutical uses of NIRS are becoming The NIR region of the spectrum contains overtones
more common. Literature references pertaining to and combination bands that are primarily attributed to
pharmaceutical uses date back to the early 1980s.[9,10] hydrogen vibrations (OH, CH, NH). These overtones
However, earlier NIR articles exist and were not taken and combination bands are much weaker than the fun-
seriously by the pharmaceutical industry at the time. In damental vibrations, thus, the molar absorptivities are
1966, Sinsheimer and Keuhnelian[11] reported quanti- much smaller than those of the corresponding infrared
tative NIRS work with pharmaceutically active com- bands. Smaller molar absorptivities allow the use of
pounds pressed into pellets. undiluted samples and penetration of solid samples
NIRS involves the multidisciplinary approaches of with good results.
the analytical chemist, statistician, and computer There are several notable differences among the
programmer. The word chemometrics refers to the near-infrared region and other infrared regions of the
application of mathematical or statistical methods to electromagnetic spectrum. Conventional infrared
measurements made on chemical systems of varying instruments usually operate in the near-, mid-, or far-
complexity. Chemometrics is defined[12] as the chemical infrared regions, depending on the energy source and
discipline that uses mathematical, statistical, and other the detectors used. The wavelength range used for
methods that apply formal logic to design or select the NIR is just beyond the visible end of the spectrum
optimal measurement procedures and experiments, and and is referred to in terms of nanometers. Other
to provide maximum relevant chemical information by regions of the spectrum are referred to in terms of wave
analyzing chemical data. numbers. Thus, the near-infrared region is 14,300–
Chemometrics has found widespread use in the 4000 cm
1, the midinfrared range is 4000–200 cm
1,
interpretation of analytical data and is relied on for and the far infrared is from 200–10 cm
1.
the development of NIRS methods.
In pharmaceutical applications, NIRS is more often
used as a secondary analytical technique than as a pri- INSTRUMENTATION
mary tool. As a secondary method, a reference method
is required to determine the reference component NIRS instruments are typically designed as either
values that are to be used in the NIR calibration. transmittance or reflectance, with some allowing the
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-100000440
3630 Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved.
Tablet Evaluation Using Near-Infrared Spectroscopy 3631

user to switch from one to the other. The difference resolution (in certain regions) among instruments. One
between the two instrument configurations lies in the of the manufacturers, Analytical Spectral Devices, Inc.,
positioning of the sample and the detector(s). In trans- Boulder, CO, was identified in the study as the source
mittance mode, the sample is placed between the of the donated spectra for the report and also provided
monochromator and the detector so that the entire their data over the Internet.
pathlength of the sample is integrated into the measure-
ment. Transmittance measurements require higher
frequency energy (800–1400 nm) because of the greater DATA ANALYSIS AND CALIBRATION
depth of penetration into the sample.[14] In reflectance
mode, the monochromatic light is illuminated directly Obtaining Sample Spectra
onto the sample, and the reflected light is collected by
detectors positioned at 45 angles to the sample. The process of scanning a sample is quite simple and
Advances in instrumentation have resulted in a wide very rapid. The sample holder and surface must first
array of choices for analysts. These include instru-

Tablet–Tablet
be gently cleaned of debris and a reference (usually
ments for specific applications with custom sample an internal ceramic Coor’s standard) scan taken. The
holders as well as general multiuse types. In the past sample may then be placed in the sample holder, which
10 years, various patents have been issued for sample may hold one or more of a specific type of sample. The
holders,[15] sample supports,[16] and a fiber optic system sample is positioned, the lid closed, and the scan taken.
for dissolution.[17] Scan times are usually approximately 40 s. If multiple
An extensive product review of recent NIR tech- scans of the same sample are needed, the sample may
nology was published by Noble.[18] Enormous progress be removed and rescanned. Instrument software facil-
has been achieved in chemometrics and computing itates the process and allows the spectra to be named
power, making many new applications possible. There and stored in data files.
are dozens of manufacturers of NIR spectrophoto-
meters in the United States. There are many vendors
of sampling components and software packages for Calibration
data analysis. Research data of the most recent instru-
mentation and software are available directly on the Calibration models are developed to determine the
World Wide Web, as most manufacturers maintain a relationship between calibration set spectra and the
Web site. There are numerous Internet[19] sites that constituent value of interest for those samples. Cali-
provide links to professional spectroscopy societies, bration involves taking spectra from many samples
publishers of spectroscopy journals, and patents[20] varying over the measurement range and also measur-
related to pharmaceutical uses of NIR. ing the desired parameters. A rugged chemometric
The US FDA’s Division of Drug Analysis spon- model for a complex sample may require hundreds to
sored a cooperative study among manufacturers of thousands of samples taken from all possible situa-
NIRS instruments and pharmaceutical manufacturers tions, in and out of specification, that it may encoun-
to analyze a series of tablets, hard and soft gelatin cap- ter. Samples selected for calibration must contain all
sules, and powders. One goal of the study was to dem- of the variables affecting the chemical and physical
onstrate that a near-infrared scan was not unique properties of the samples to be analyzed. To character-
among manufacturers. Another reason for the study ize each source of variation, it is recommended that 15
was to establish FDA guidelines for the Instrument to 20 samples be run for each variable. Application of
Qualification and Performance Verification of NIR a math treatment, such as second derivative, prepares
instruments used in pharmaceutical analysis. Nine the raw spectral data for use in a regression and sub-
manufacturers participated in the study and repre- sequent development of a calibration equation. This
sented makers of numerous types of monochromators type of treatment results in a data file that will yield
(acousto-optical tunable filter or AOTF, dispersive and more information more easily than a raw data file.
Fourier transform-NIR or FT-NIR). Ciurzcak[21] Because NIR bands are mixtures of overtones and
reported a detailed account of the study to compare combinations, the intensity of the absorbance at
instrument performance in similar products, thus pro- particular wavelengths do not necessarily respond
viding some comparative information to potential linearly to a change in concentration. In the case of a
buyers of NIR equipment. Ciurczak concluded that mixture, band mixing may further disrupt any linear
the quality of the spectra depends on the mono- relationship between the intensity and the concen-
chromator, and there is a range of noise variability tration. These are the reasons why the simple
in both FT-NIR and dispersive-type instruments. application of Beer’s law A ¼ ebc, where
Examination of the spectra appearing in the report A ¼ absorbance, e ¼ absorptivity, b ¼ path length,
indicated differences in photometric noise and in spectral and c ¼ concentration to NIR bands may not generate
 3632 Tablet Evaluation Using Near-Infrared Spectroscopy

equations suitable for quantitation. Multiple a longer period (hours or days). Repeatability refers to
regression techniques help avoid this problem. Linear the instrument’s ability to generate consistent measure-
calibration methods such as multiple linear regression ments under the same conditions (without removing
(MLR), principal component analysis (PCA), and the sample from the instrument) over a relatively short
partial least-squares regression (PLS) are routinely period (seconds or minutes). All of these factors must
used in NIR analyses. The choice of regression tech- be addressed to ensure the validity of the NIR
nique is subjective, depending on many factors. For calibration model.
further description of these techniques, see the
introduction to multivariate calibration methods by Regulatory Issues
Thomas.[22] Other chemometric techniques useful in
NIRS methods include BEAST bootstrap error-adjusted The American Society for Testing and Materials
single-sample technique (BEAST), a non-parametric (ASTM) recently published an official document[27]
clustering algorithm used by Lodder and co-workers[23] providing a guide to spectroscopists for the multi-
to detect capsule tampering by NIRS. Qualitative NIR
Tablet–Tablet

variate calibration of infrared spectrometers. The scope

methods use pattern-recognition techniques to ‘‘train’’ of the publication, entitled Standard Practices for
the computer to identify an unknown material. Residual Infrared Multivariate Quantitative Analysis includes
variance methods and discriminant analyses (Mahalano- a description of multivariate calibration methods for
bis distance) were described by Mark and Tunnell,[24] the determination of physical or chemical characteris-
and were compared by Gemperline and Boyer.[25] tics of materials. This document is the first official stan-
Gerhausser and Kovar[26] reported strategies for optimi- dard for the application of chemometric multivariate
zation of spectral libraries and compared two pattern- analysis to near-infrared and infrared instruments.
recognition methods to identify 117 drug substances. Regulatory bodies around the world have given
approval for NIR methods for a variety of purposes.
In June 1995, the Medicines Control Agency (MCA)
Validation granted approval to Glaxo Wellcome in the United
Kingdom for a NIR method for the identification
Regardless of the mathematics used to interpret NIR and assay of ZoviraxÕ (acyclovir) tablets. This is
spectra, the method must still undergo a validation believed to be the first official approval for NIR
process. The principal elements of ensuring linearity, granted by the MCA as an assay method for tablets.
accuracy, selectivity, and reproducibility of a quanti- In Norway, Wieders Farmasoytiske A/S received
tative method are required. approval for the use of NIR as an alternative method
The validation process determines the amount of for identification, assay, and determination of moisture
error owing to variation among the values in the popu- content of paracetamol (acetaminophen) tablets. The
lation. It is used to check for the existence of a relation- Norwegian Medicines Control Authority approved
ship between the calibration set and the validation set. the method in December 1996.
Manufacturers of NIRS instrumentation include Other regulatory approvals are described in later
software packages that allow the operator to predict sections of this chapter.
analytical results on data files that have been stored,
thus allowing for validation of the calibration equation
and testing for errors in the developed calibration. CURRENT METHODS OF
This enables calibration equation performance testing TABLET EVALUATION
in terms of precision. The validity of these models
depends on the ability of the calibration set to accu- Official standards for the evaluation of tablets are
rately represent the samples in the prediction set. given by the U.S. Pharmacopeia (USP) and other com-
One source of prediction error is the inherent accu- pendia and include uniformity of dosage units (weight
racy and precision of the reference analytical method variation, content uniformity) and disintegration test-
used. If the reference method produces erroneous ing. Unofficial tests include those for mechanical
values that are consistently high or low, this bias will strength (hardness, crushing strength) and resistance
be reflected in the prediction results. Other sources of to abrasion (friability). A major disadvantage of cur-
prediction error relate to the reproducibility, stability, rent compendial methods of tablet evaluation is that
and repeatability of the NIR instrument. Reproduc- they are time-consuming and destructive in nature.
ibility (precision) is validated by making repeated mea- Once a test is performed on a sample, the integrity of
surements of the same sample and removing it between that sample is usually lost (with the exception of weight
runs. Small changes in conditions may occur owing to testing) and no additional testing may be done on it.
multiple insertions of a sample onto the instrument. For example, a traditional quantitative analysis such
Stability refers to similar changes that may occur over as high-performance liquid chromatography (HPLC)
Tablet Evaluation Using Near-Infrared Spectroscopy 3633

typically calls for the tablet to be ground, followed allowing a direct correlation between tests. Economic
by dissolution and dilution in a suitable medium. A benefits are obvious for manufacturers, who may
significant amount of time and labor is required increase profits per batch because of the need for fewer
to run each sample in duplicate or triplicate. Other retained samples. NIRS is particularly useful in the
traditional methods of tablet evaluation also involve early stages of product development when the supply
time-consuming sample preparation, such as KBr of the new drug is limited.
dilution for mid-infrared analysis. NIR analysis is rapid, requiring less than one minute
The mechanical strength of a tablet plays an impor- to analyze a single sample. Also, NIR analyses do not
tant role in the development and control procedures. require the use and ultimate disposal of organic solvents,
Crushing strength is the most widely used test of mech- thereby reducing environmental waste. Advances in
anical strength. It is defined as the compression force instrumentation, fiber optics, and software offer many
that, when applied diametrically to a tablet, just frac- options. Portable NIRS units are not uncommon.
tures it.[28] Tablet hardness depends on the weight of Accelerated degradation samples may be analyzed
material and the space between the upper and lower and returned to the associated storage condition at

Tablet–Tablet
punches at the moment of compression. Inconsistent each of the appropriate time intervals, thus drastically
hardness values are likely to result from variation in these reducing the number of samples taken from the batch.
parameters. The fundamentals of powder compression Measurement of powders can take place directly
are given in a report by Leuenberger and Rohera.[29] through the unopened glass jar or vial because glass
The Erweka hardness tester measures horizontal does not absorb in the NIR region. Direct measurement
crushing strength by applying a load at 90 to the long- through the container further reduces sampling errors,
est axis. This type of hardness tester is subject to two which may be introduced when a sample is withdrawn.
sources of inherent error: 1) the possibility of an incor- Multiple components of a sample may be analyzed
rect zero and 2) a scale that does not accurately indi- simultaneously. A single spectrum can be obtained and
cate the true load applied. Other commercially used compared with several different calibration sets at the
instruments include the Strong-Cobb, Monsanto, and same time, allowing the measurement of several constitu-
Pfizer hardness testers. Variations in crushing strength ents at one time. This saves considerable time and labor.
values obtained from different types of hardness testers
may be attributed to inaccuracies in instrument scale
values, incorrect zero adjustment, and varying methods LIMITATIONS OF NIRS
of applying the load. This necessitates calibration when TABLET EVALUATION
comparing results from different types of testers. The
physical dimensions and shape of the tablet may also The initial calibration process for a substance or a pro-
contribute to the property of crushing strength. duct can be quite detailed. A calibration equation is
The conventional methods of hardness testing for needed for each constituent in the sample. NIR cali-
tablets also involve a subjective operator error. The scale brations must be formulation-specific. The accuracy
on the Erweka hardness tester is divided into segments of of the NIR method cannot be better than the reference
0.25 kg units. Very often, the sample under evaluation method from which it was built. Ruggedness of the
may produce a reading that falls between two divisions, models improves when all expected types of variation
and it is up to the operator to decide on the result. are included in the model. Careful selection of
Tablet-coating processes are commonly used in the representative samples is imperative to the successful
pharmaceutical industry. Aqueous film coatings com- performance of the calibration. The choice of math-
posed of cellulose derivatives and other polymers are ematical models depends highly on the character of
useful for the control of dissolution of drug from the the sample.
tablet. Gravimetric analysis and HPLC are often used Another issue is that of transferability of the cali-
to determine the endpoint of the coating process. bration model among instruments. This has been a
significant obstacle to more widespread use of NIR
methods. Transferability is especially important to
ADVANTAGES OF NIRS multisite facilities, because it is needed to avoid time-
TABLET EVALUATION consuming recalibration procedures. Calibration errors
may occur among instruments because of slight differ-
NIRS is a non-destructive method, thus, 100% inspec- ences in instrument response, especially if full-spectrum
tion of batches is theoretically possible, allowing multivariate models are used. Shenk and Westerhaus[30]
better control of product uniformity. NIR is also addressed the problem and proposed a standardization
non-invasive, enabling subsequent evaluation of the algorithm, which was modified by others.[31,32]
same tablets by another method. Samples may be Physical attributes of the tablets can affect the cali-
retained for further analysis by NIR or other methods, bration process. For example, scored tablets and those
 3634 Tablet Evaluation Using Near-Infrared Spectroscopy

of differing geometries may produce more variable spectra with the library of a representative collection of
NIR spectra than flat, unscored tablets. Work in our approved batches of the same material.
laboratory[33] demonstrated a difference in NIR hard-
ness testing of scored AvicelÕ-based chlorpheniramine
Particle Size
tablets. Mixed calibration models composed of flat and
concave tablets gave variable hardness prediction
The measurement of particle size is a key issue in the
results, supporting the assertion that calibration models
formulation of many pharmaceutical products. Particle
should contain samples of homogeneous composition.
size distribution is known to directly influence physical
Borer and coworkers[34] published a useful evalua-
properties of powders, such as dissolution rate, powder
tion of sources of variability encountered in the NIR
flow, bulk density, and compressibility. Conventional
analysis of drug products. Analysts involved in NIRS
methods of particle size measurement include sieve
method development should consider this report.
analysis and laser diffractometry.[39]
Parameters included in the study included.
Ciurczak and associates[40] reported a NIR method
Tablet–Tablet

of determination of particle size of pure, granular sub-

1. instrument settings (number of scans averaged
stances. The method is based on theories of reflected
per spectrum, iris opened or closed, frequency
light, in which reflectance increases as the particle size
of reference spectrum collection);
decreases. The reference method was a low-angle laser
2. data treatment (segment value used for second-
light-scattering (LALS) particle sizer (Malvern). The
derivative calculation); and
researchers found linear results for particles above
3. library design (total number of samples, number
85 mm, but less accurate results for smaller particles.
of days required to complete the library).
O’Neil, Jee, and Moffat[41] described a NIRS
method for measuring the median particle size of
numerous compounds and excipients. Malvern data
BULK DRUG PROPERTIES
and NIR reflectance spectra of sieved fractions and
bulk samples were used to construct multiple linear
Raw Material Identification
regression (MLR) and quadratic least-squares fit cali-
bration models. The same group[42] reported success
NIRS is useful for the analysis of both raw materials
in measuring the cumulative percentage frequency par-
and finished dosage forms. Qualitative determination
ticle size distribution of microcrystalline cellulose
of pharmaceutical raw materials using NIRS was
(Avicel) using NIRS. This study compared the use of
reported as early as 1982.[35] The largest variations in
three-wavelength MLR and principal component
commercially produced excipients and actives appear
regression (PCR), each model using Malvern and NIR
to be in moisture content and particle size.[36,37] These
reflectance data. The PCR model produced smaller
parameters may be monitored by NIRS with relative
SEPs, suggesting a more robust model than the MLR.
ease. Incoming substances may be tested for immediate
Frake and co-workers[43] extensively evaluated
identity confirmation on the receiving dock by insert-
numerous chemometric techniques for the NIRS pre-
ing a fiber optic probe directly into the barrel. Spectral
diction of mass median particle size determination of
data for each lot of material purchased may be saved
lactose monohydrate. Models evaluated in zero order
and added to the growing spectral reference library.
(untreated) and second derivative were MLR, PLS
The use of NIRS as an identification method
(partial least squares), and ANN (artificial neural net-
involves the establishment of a comprehensive library
work). The researchers concluded that there is more
containing the spectra of many compounds. The sample
than one way to treat data and achieve a good cali-
is scanned and identified by finding the most similar item
bration model. The group also confirms previous
in the library. Comparison of the sample spectrum to the
observations that derivitization of data does not
library reference spectrum is achieved by calculating (via
remove ‘‘particle size effects’’ (previously thought to
computer) the cosine of the angle between the vectors of
contribute to baseline shift).
both spectra. This value is called the spectral match
Other researchers[44] have also reported NIR par-
value (SMV) and may range from
1 to þ1, with þ1
ticle size studies.
being a perfect match. Plugge and Van Der Vlies[38] uti-
lized the SMV for a NIR method of identification of
ampicillin trihydrate. In their work, an SMV of 0.9980 Polymorphism and Racemization
was established as a minimum value for positive identity
of ampicillin. The conformity index or CI is also calcu- Many drugs have the ability to form several dis-
lated to establish the degree of conformity of a batch tinct crystalline forms or polymorphs. Although the
to specifications. Conformity testing detects deviations polymorphs are chemically identical, they are different
from normal processing by comparing the average batch arrangements of molecules and may exhibit different
Tablet Evaluation Using Near-Infrared Spectroscopy 3635

properties. Each polymorph may possess different energy described a NIRS method to determine at-line moist-
levels and ultimately affect the dissolution rate of the ure content in bulk hard gelatin capsules. Capsules
compound. The infrared spectra of polymorphs may be were equilibrated at various humidities, and reference
expected to vary owing to different arrangement of moisture content was determined by loss on drying.
functional groups (hydrogen bonding and polarization Zhou and co-workers[55] at Eli Lilly and Company
may be affected). Traditional methods for the identifi- developed a NIRS method to determine moisture
cation of polymorphism are differential scanning content in a freeze-dried drug product using gas chro-
calorimetry (DSC) and X-ray crystallography. However, matography (GC) as the reference method. A standard
these technique are destructive, and thus multiple runs of error of prediction (SEP) as low as 0.07% w/w was
the same sample are not possible. Ciurzcak[45] reported a obtained using the NIR method in the range of
NIR method of identification of the polymorphs of 0.1–5.7% water. At very low water levels, the workers
caffeine. Aldridge and co-workers[46] reported a NIR found the GC method to be more precise than the
method of detection of polymorphism in which pattern- NIR method.
recognition methods were used to discriminate between Corti and co-workers[56] developed a NIRS method

Tablet–Tablet
the desired polymorphic form of a drug substance and to determine the water content of crushed ranitidine
another undesired polymorph. A significant outcome of chlorhydrate tablets. KF titration was used as the ref-
this work was the successful transfer of the calibration erence method. Higher NIRS errors were found with
between six other NIR instruments without the use of the samples having a water content of less than 1%.
multivariate calibration transfer algorithms. Dziki and coworkers[57] used NIRS to monitor the
Gimet and Luong[47] used NIRS to determine mobility of water within the sarafloxacin crystal lattice.
whether the processing of a granulation resulted in The study involved the veterinary product sarafloxacin
racemization. Others[48] have also reported the differ- in an aqueous granulation process. A failed lot of the
ential identification of optically active forms. product was indistinguishable from an acceptable lot
using X-ray powder diffraction, midinfrared spec-
troscopy, differential scanning calorimetry (DSC),
Moisture Content and TGA. NIRS detected intermediate stages of water
absorption in the granulation and enabled the process
The classic methods of water determination are Karl to be controlled.
Fischer (KF) titration, gas chromatography (GC), and An interesting report on NIR measurement of water
loss on drying (LOD). Using near-infrared methods, in skin was published recently by Martin[58] at Helene
the presence of water is indicated by a major NIR Curtis, Inc. NIRS measurements indicated four types
absorption band at approximately 1920–1950 nm. The of water contained in the skin. The workers presented
absorption maximum and peak shape depend on tentative assignments for the absorption bands:
the degree of hydrogen bonding occurring within the
environment where the water is located. The stronger  lipid bilayers (1875 nm),
the hydrogen bond, the longer the wavelength of the  secondary and primary water on protein groups
NIR absorption maxima. A second band attributed (1909 and 1923 nm), and
to water may appear at approximately 1450 nm and  bulk water beneath the stratum corneum (1890nm).
is attributed to bound water.
The NIRS determination of moisture content has Studies were performed in vivo on two volunteers
been documented by numerous researchers.[49–51] Plugge as well as in vitro using porcine skin. Although not
and Van Der Vlies[52] described a NIR method for ampi- directly related to tablet evaluation, the work is
cillin, which measures eight quality control criteria: notable for its contribution to the transdermal delivery
of drugs, potential NIR prediction of drug penetration
 identity, through the skin, and conclusions regarding moisturi-
 water content, zer use on the skin.
 crystallinity,
 ampicillin content,
 fraction of anhydrous ampicillin, PRODUCT EVALUATION
 residual reagent,
 residual organic solvents, and Identification and Potency
 residual starting material.
Reports describing NIR methods of identification of
DeThomas and VonBargen[53] patented a NIR fiber solid and liquid dosage forms have increased in the
optic system for the measurement of moisture or ‘‘a literature during the 1990s.[59–62] Earlier work has been
constituent’’ in a powder. Berntsson, and associates[54] reviewed elsewhere (see Introduction). Virtually all
 3636 Tablet Evaluation Using Near-Infrared Spectroscopy

new NIR methods include product identification in Identity testing of blister packaged
the assay. Gottfries and co-workers[63] reported a tablets/supplies for clinical trials
NIR method for the measurement of metaprolol in
controlled-release tablets. In their work, a comparison A non-invasive NIR method was reported by Dempster
was made between the diffuse reflectance and trans- et al.[70] for the identification of tablets within blister
mission modes. The workers found better prediction packages. The method identified and discriminated
of tablet strength using transmission mode and rea- various potencies of an experimental drug, placebo
soned that reflectance spectra are more sensitive to tablets, and clinical comparator tablets.
the inhomogeneity of the material. Aldridge and coworkers[71] described a NIRS
Ebube and coworkers[64] reported a NIR method method for non-destructive identification of blister-
that can distinguish between three AvicelÕ products packaged tablets. The NIR method drastically reduced
owing to the varying particle sizes. The method was the assay time required by the previous TLC method.
also designed to predict tablet hardness and lubricant The TLC method required a full day to analyze 40
concentration. Regardless of the small sample sizes tablets, compared with the NIR method, which
Tablet–Tablet

used, good results were obtained. analyzed 10 tablets in 7 min.

North, Young, and Leng[65] recently presented an
interesting comparison of NIR diffuse reflectance and Detection of adulteration
transmittance for the analysis of tablets. Tablets used
were of different drug content and size as well as Lodder, Selby, and Hieftje[72] reported a NIR method
coated and uncoated. Both methods produced excel- of detection of capsule tampering. Several non-
lent resolution of two tablet strengths. Uncoated and prescription products were selected for the study and
coated tablets produced different spectra by reflec- tested with and without the addition of various
tance, but only a minimal difference was seen by trans- adulterants. The NIR method provided a rapid, non-
mission. Of two drug peaks seen in the reflectance invasive way to screen products for known foreign
spectra, only one band was observed in transmission substances. One limitation of the NIR method is the
because no transmission through the tablet occurred inability to predict which substances might be present
in the region of the second peak. Both methods pro- in a product. Although the NIR spectra of many adul-
duced difference in spectra as a result of difference in terants may be present in the spectral library, there is
tablet size and surface curvature. the possibility that a new, unknown substance could
Merckle and Kovar[66] reported an assay of effer- be added to a product and not detected as an error
vescent tablets (intact and powdered) by NIRS in in the NIR product spectrum.
transmittance and reflectance modes. Results of quan-
titative determination of acetylsalicylic acid (ASA) and Product development
ASA in combination with ascorbic acid and/or parace-
tamol were comparable in both transmittance and In the drug-discovery field, chemists can use NIR to
reflectance modes. Corti and associates[67] described a monitor the progress of reactions. Hearn and co-
NIR transmittance analysis of coated tablets, using workers[73] reported a NIR method to monitor the
both whole and milled tablets. preparation of compounds for screening as antituber-
Brashear and coworkers[68] developed a diffuse culosis drugs. The reaction of isonicotinic acid
reflectance NIRS method to quantify lomefloxacin and hydrazide (INH) with carbonyl compounds in the
polyethyleneglycol (PEG) within a polymeric implant. preparation of Schiff bases was followed. Forbes,
The properties of pore-forming excipients such as Persinger, and Smith[74] described a NIR conformance
PEG are known to affect the rate of drug release from test method to assay and identify two chemical inter-
a matrix. mediates used in the manufacture of Loracarbef, a
Sondermann and Kovar[69] described a study using carbacephalosporin.
NIRS for the identification of ‘‘ecstasy’’ street samples. Bauer, Dziki, and Quick[75] used NIRS to investigate
Ecstasy tablets may contain either N-methyl-3,4- the problem of dissolution failure in an erythromycin
methylendioxyamphetamine (MDMA) or N-ethyl-3,4- tablet formulation. The technique enabled the group
methylendioxyamphetamine (MDE) as well as other to identify the presence of a dehydrated dihydrate pro-
amphetamine derivatives. In addition, various excipients duced during formulation. The dihydrate was found to
were present, and non-standardized production pro- gradually bind with magnesium hydroxide in the tablet
cedures contributed to inhomogeneous tablets. The formulation, thus delaying the process of dissolution.
authors researchers included a broad range of excipients The use of NIR facilitated the development of a humidi-
in their calibration work and succeeded in constructing fying process that reversed the binding and increased the
three PLS models for identification. dissolution rate.
Tablet Evaluation Using Near-Infrared Spectroscopy 3637

Quality Control Parameters in spectra to the mass of water absorbed, the mass of
salicylic acid formed, and the time the tablets spent
Tablet hardness in a hydrator.
Shimoyama and coworkers[84] reported a NIR
It has been shown[76,77] that the NIR signal varies with analysis of photodegradation of poly(methyl meth-
a change in compression force. In other words, changes acrylate) using an in situ fiber optic device. This type
in tablet hardness result in an alteration in the NIR of technology from a related discipline is notable as a
spectra of the sample. Presumably, increasing the com- potential application for pharmaceutical systems.
pression force during the tableting process causes the
tablet to be smoother as well as harder, thus causing Characterization of powder blends
less light scattering, leading to a greater absorbance and blend homogeneity
and higher baseline. NIR hardness testing of tablets
can be performed at the same time as other parameters Powder blending is a fundamental step in the process
such as identity, moisture content, and coating thickness. of manufacturing pharmaceutical products. Only a

Tablet–Tablet
Further work by Kirsch and Drennen[78] on NIR homogeneous mixture can be properly subdivided to
hardness testing has explored the use of various math- provide uniform doses of the active ingredients. The
ematical models for calibration. Other researchers have current procedures for monitoring blend uniformity
also reported NIR methods for the measurement of require that the blending process be stopped at defined
tablet hardness.[64,79] intervals to obtain samples of the blend. The samples
are collected from different locations in the blending
Tablet coating vessel using a sample thief. The samples are then sent
to a laboratory and analyzed using traditional methods
NIRS has been used to determine tablet-coating and such as HPLC or UV until the active components are
core thickness. Kirsch and Drennen[80] evaluated the within specification for that formulation. This approach
use of NIR at-line to monitor film coating in a Wurster requires a significant amount of time and labor and may
column. The method was successful at predicting coat- be subject to errors induced by sampling methods.
ing thickness of two coating formulations at various NIRS has been shown by several researchers to be
intervals during the process and was less time-consuming useful for evaluating the powder-mixing process. Wargo
than wet chemical methods. Earlier work by Kirsch and and Drennen[85] demonstrated the use of a NIR method
Drennen[81] described a NIR method to determine film- to determine homogeneity of powder blends.
coating parameters of theophylline tablets. Increasing In 1995, the European Patent Office granted a
coating thickness, corresponded to increased NIR absor- patent to Dr. Paul K. Aldridge, Pfizer Central
bance in certain regions of the spectrum. Calibration Research, Groton, CT, for an apparatus[86] for mixing
models were developed for tablet hardness, coating and detecting on-line homogeneity. The apparatus
thickness and the prediction of time to 50% dissolution. involves the use of a diffuse-reflectance fiber optic
Buchanan and coworkers[82] at Merck reported a probe interfaced on-line with a V-blender. Sekulic
NIR method for evaluating a new coating-thickness and co-workers[87] at Pfizer described the use of this
manufacturing process. A precision film-coating process apparatus for on-line monitoring of powder blend
was tested whereby an immediate-release drug-active homogeneity. An 8-quart twin-shell V-blender was
coating surrounded an extended-release active-drug interfaced with a fiber optic probe at the axis of
core. The NIR method enabled the evaluation to rotation. Spectra were collected at prescribed intervals,
proceed more quickly and less expensively than did the and data analysis was performed using a series of
reference HPLC method. The implementation of the commercial software. Variability in the NIR spectra
NIR method allowed rapid evaluation of tablets and as a function of time was measured, and it was shown
assisted in identifying ‘‘dead zones’’ in the Wurster that this variability reached a minimum level sooner
column, thus allowing immediate correction and revision than what traditional blending times suggest.
of the process. DeMaesschalck et al.[88] used the NIR on-line method
described in article by the Sekulic and associates to
Determination of degradation products design an approach for deciding when the blend is
homogeneous. They calculated the average standard
Drennen and Lodder[83] reported a non-destructive deviation between spectra taken at each time and used
NIRS method to monitor the decomposition of the dissimilarity between each new measurement and
aspirin. In contrast to the multi step HPLC assay for the ideal mixture spectrum to monitor changes in the
salicylic acid and the USP identity tests for aspirin, mixture during the blending process.
the NIR method involved a 90 s scan of individual Scientists at Merck were issued a U.S. patent in
intact aspirin tablets. The workers correlated changes 1996 for a method[89] of measuring the homogeneity
 3638 Tablet Evaluation Using Near-Infrared Spectroscopy

of tablets using NIR. It can be used to monitor the 7. Workman, J.J., Jr. Review of process and non-invasive
pharmaceutical material during the tableting process near-infrared and infrared spectroscopy. Appl. Spectrosc.
Reviews, May 1 1999, 34, 1–2, 1–89.
(powder mix, granular mix, and compressed tablets). 8. Ciurczak, E.W.; Drennen, J.K. Near Infrared Spectroscopy
in Pharmaceutical and Medical Applications, Practical
Spectrosc. Series; Gadamasetti, K.G., Ed.; Marcel Dekker,
Inc.: New York. In press.
CONCLUSIONS 9. Rose, J.J.; Prisick, T.; Mindakia, J. J. Parent. Sci. Tech.
1982, 26, 71–78.
NIRS has proven to be a fast, reliable, and cost-saving 10. Ciurczak, E.W.; Torlini, R.P. Analysis of solid and liquid
dosage forms using near-infrared reflectance spectroscopy.
method for numerous applications in the pharmaceuti- Spectrosc. 1987, 2 (3), 41–43.
cal industry. It is no longer the esoteric method it was 11. Sinsheimer, J.E.; Keuhnelian, Am. J. Pharm. Sci. 1996, 55,
once believed to be. The pharmaceutical industry has 1240.
12. Massart, D.L.; Vandeginste, B.G.M.; Deming, S.N.;
learned a great deal about NIRS from the agricultural Michotte, Y.; Kaufman, L. Data handling in science and
and food industries. Concepts and techniques have technology. In Chemometrics: A Textbook; Vandeginste,
been borrowed and fitted to the needs of pharmaceuti- B.G.M., Rutan, S.C., Eds.; Elsevier Publishing: New York,
Tablet–Tablet

1988; Vol. 2, 5.
cal scientists. Users in all disciplines face common 13. Willard, H.H., et al. Infrared spectroscopy. In Instrumental
issue, such as calibration transfer, moisture contami- Methods of Analysis, 7th Ed.; Wadsworth Publishing
nation, particle size, and the rigors of calibrating mul- Company: Belmont, 1988; 287.
14. Workman, J.J., Jr.; Burns, D.A. Commercial NIR instru-
tiple constituents. mentation. In Handbook of Near-Infrared Analysis; Burns,
NIRS possesses a great and, as yet, incompletely D.A., Ciurczak, E.W., Eds.; Marcel Dekker, Inc.: New
exploited potential in the area of identity testing of York, 1992; 37–51.
15. Lodder, R.A. Sample Holders or Reflectors for Intact
drug substances. It has already begun to replace Capsules and Tablets and for Liquid Microcells for Use in
traditional compendial methods of quality control. It Near-Infrared Reflectance Spectrophotometers. US Patent
has gained recognition from the FDA. and other regu- 165,751, November 21, 1989.
16. Drennen, J.K. Near-Infrared Reflectance Spectrometer
latory agencies, a signal to skeptics that NIRS is a solid System and Related Sample Cell and Sample Support.
alternative to traditional methods of analysis. Aggres- US Patent 898,454, May 25, 1993.
sive workers in the field are moving to develop and 17. Soloman, S. Non-Destructive Identification of Tablet and
Tablet Dissolution by Means of Infrared Spectroscopy.
receive approval for NIR methods that bypass the tra- US Patent 338,909, October 21, 1997.
ditional reference methods. A greater understanding of 18. Noble, D. Illuminating near-IR. Anal. Chem. 1995, 67 (23),
the mathematics involved with NIR analyses has 735A–740A.
19. http://kerouac.pharm.uky.edu/asrg/cnirs/ir_spec.html
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companies that wish to use NIR analyses may find it 20. http://leden.tref.nl/~mderksen/patents.html (accessed May
wise to contract out their work to groups with more 2000).
21. Ciurczak, E.W. Pharmaceutical mixing studies using near-
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Tablet Formulation
Larry L. Augsburger
Department of Pharmaceutics, University of Maryland at Baltimore,
Baltimore, Maryland, U.S.A.

Mark J. Zellhofer
Guilford Pharmaceuticals, Inc., Baltimore, Maryland, U.S.A.

Tablet–Tablet
INTRODUCTION 5. Manufacturability. The formulation design should
allow for the efficient, cost-effective, practical pro-
The best new therapeutic entity in the world is of little duction of the required batches.
value without an appropriate delivery system. Tableted
drug delivery systems can range from relatively simple That tablets can be formulated to uniquely meet
immediate-release formulations to complex extended- these criteria accounts for their emergence as the most
or modified-release dosage forms. The most important prevalent oral solid dosage form. Although several
role of a drug delivery system is to get the drug ‘‘deliv- different types of tablets may be distinguished, they
ered’’ to the site of action in sufficient amount and at are mostly made by compression, intended to be swal-
the appropriate rate; however, it must also meet a lowed whole and designed for immediate release. This
number of other essential criteria. These include physi- paper presents a systematic approach to the design and
cal and chemical stability, ability to be economically formulation of immediate-release compressed tablets.
mass produced in a manner that assures the proper
amount of drug in each and every dosage unit and in
each batch produced, and, as far as possible, patient
MODERN TABLET FORMULATION DESIGN
acceptability (for example, reasonable size and shape,
AND MANUFACTURE
taste, color, etc., to encourage patients to take the drug
and thus comply with the prescribed dosing regimen).
Tablet dosage forms have to satisfy a unique design
The discovery of new therapeutic entities always
compromise. The desired properties of rapid or con-
initiates excitement, but the contributions of the for-
trolled disintegration and dissolution of the primary
mulation specialist are either not well understood or
constituent particles must be balanced with the manu-
are often taken for granted and thus remain ‘‘unsung.’’
facturability and esthetics of a solid compact resistant
However, the drug and its delivery system cannot be
to mechanical attrition.
separated. The general design criteria for tablets are
Excipients are critical to the design of the delivery
given as follows:
system and play a major role in determining its quality
and performance.[1] They may be selected to enhance
1. Optimal drug dissolution and, hence, avail- stability (antioxidants, UV absorbers), optimize or
ability from the dosage form for absorption modify drug release (disintegrants, hydrophilic poly-
consistent with intended use (i.e., immediate or mers, wetting agents, biodegradable polymers), provide
extended release). essential manufacturing technology functions (binders,
2. Accuracy and uniformity of drug content. glidants, lubricants), enhance patient acceptance (fla-
3. Stability, including the stability of the drug vors), or aid in product identification (colorants). Thus
substance, the overall tablet formulation, dis- a tablet formulation is not a random combination of
integration, and the rate and extent of drug ingredients, but rather a carefully thought out, rational
dissolution from the tablet for an extended formulation designed to satisfy the above criteria.
period. A long list of possible excipients is available to the
4. Patient acceptability. As much as possible, the formulation scientist, but certain external factors
finished product should have an attractive such as cost, functional reliability, availability, and
appearance, including color, size, taste, etc., as international acceptance govern their selection. For
applicable, in order to maximize patent example, although the official compendia provide
acceptability and encourage compliance with standards for identity and purity of excipients, mono-
the prescribed dosing regimen. graphs may not provide tests to assure their functionality.
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-100001725
Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved. 3641
 3642 Tablet Formulation

For instance, the NF monograph for Compressible (a previously new chemical entity whose patent has
Sugar provides no test for compressibility. The mono- expired and which is available to the generic market)
graph for Lactose USP does not address the many for which a USP monograph exists.
particle size and tableting grades meeting monograph
standards. The NF monograph for Pregelatinized
Identity and Purity
Starch refers to grades that are ‘‘compressible and
flowable in character,’’ but provides no specifications
The study of any drug substance must start with the
or tests for these properties. Nor do the monograph
determination of identity and purity. Such tests are
tests for disintegrants and lubricants necessarily relate
necessary to identify degradents and contaminants
to their functionality. The need to provide functional-
and may include organoleptic tests for color, odor,
ity tests or tests for properties clearly related to func-
and taste. Purity tests can be found in the USP for
tionality may be as important as controlling identity
almost all marketed compounds. Alternative methods
and purity.[2] This point has been made even more
can be employed only if they are validated against
apparent in recent years with the emergence of mul-
Tablet–Tablet

the USP procedure. Tests other than potency, which

tiple sources of such modern excipients as direct-
can help to identify or determine the purity of com-
compression filler-binders and the various classes of
pounds, are melting point, specific rotation, pH, heavy
‘‘super’’ disintegrants.
metals, residue on ignition, etc. Impurities can
A major problem currently being faced by multina-
occasionally affect stability, and metal contamination
tional firms and others who market in the international
can catalyze chemical reactions. Impurities can also
arena, is the lack of universal acceptability of excipi-
alter the color of drug substances. Techniques can be
ents in different countries. The selection of excipients
utilized to give a quantitative estimate of impurities
for international markets is often a compromise
such as the impurity index (II) and the homogeneity
between functional efficacy, local restrictions, and cost
index (HI). An ordinary impurity test can be found
and availability in the countries where the product is to
in the USP that estimates impurities by thin-layer chro-
be made. In recent years, the globalization of the phar-
matography (TLC).
maceutical industry has brought about an intense
interest in developing harmonized pharmacopeial
excipient standards, Good Manufacturing Practices Crystal Properties and Polymorphism
(GMP) for excipient manufacture, and safety evalu-
ation guidelines for new excipients to eliminate or Many drug substances appear in more than one
avoid trade barriers between different countries.[3] polymorphic form. The form is determined by certain
The International Pharmaceuticals Excipients Council conditions during the crystallization step. Occasionally
(IPEC), which consists of producers, users, and phar- drug substances are precipitated in such a way that
maceutical scientists, was launched in 1991 to assist molecules do not organize themselves in any set pat-
regulatory authorities in the United States, Japan, tern, resulting in an amorphous powder. It is also pos-
and Europe with harmonization. The separate organi- sible for solids to entrap solvents stoichiometrically to
zations later formed in the United States (IPEC- form solvates.
Americas), Europe (IPEC-Europe), and Japan (JPEC) Even though they are chemically identical, the differ-
are now known as TriPEC and include, as of 1993, ent polymorphic forms of a compound are associated
more than 100 excipient and pharmaceutical firms.[3] with different free energies, and, therefore, have different
physical properties that can impact significantly on pro-
duct performance.[4] These include differences in solu-
PREFORMULATION bility and dissolution rate (affecting bioavailability),
solid-state stability (affecting potency), deformation
The objective of preformulation studies is to develop a characteristics (affecting compactibility), and particle
portfolio of information about the drug substance to size and shape (affecting powder density and flow
serve as a set of parameters against which detailed properties). The form with the lowest energy is more
formulation design can be carried out. Preformulation stable than the others. Although the other polymorphs
investigations are designed to identify those physico- are thus energetically unfavored, if kept dry, they may
chemical properties of drug substances and excipients persist indefinitely and are called ‘‘metastable.’’ A meta-
that may influence the formulation design, method of stable form may be preferred, particularly for its ability
manufacture, and pharmacokinetic-biopharmaceutical to dissolve more rapidly.
properties of the resulting product. Polymorphic transformation can take place during
Following is a generalized preformulation protocol pharmaceutical processing, such as particle size reduc-
appropriate for tablet dosage forms. For certain tests, tion, wet granulation, drying, and even during the com-
it is assumed that the drug substance is multisourced paction process.[5] Tests employed to determine crystal
Tablet Formulation 3643

properties include differential thermal analysis (DTA),  Anticipating problems in the physical mixing of
differential scanning calorimetry (DSC), and X-ray dif- powders and the homogeneity of intermediate and
fraction.[4] See also the article Thermal Analysis of Drugs final products because significant differences in true
and Drug Products by D. Giron in this encyclopedia. densities can result in segregation,
 Anticipating problems in flow properties, since that
property is affected by density, and
Particle Size, Shape, and Surface Area
 Identifying differences in different lots and raw
materials from different suppliers because different
Probably no characteristics of a drug substance are
polymorphic forms can be expected to exhibit
more important than particle characteristics in deter-
different true densities.
mining its performance in a formulation. This is parti-
cularly true in those cases where the drug is a poorly
A comparison of true particle density, apparent
soluble non-electrolyte or a free acid form with poor
particle density, and bulk density can provide infor-
solubility at low pH values. Such drugs are likely to
mation on total porosity, interparticle porosity, and

Tablet–Tablet
exhibit dissolution-rate-limited absorption, and if
intraparticle porosity. Methods include true particle
dissolution does not take place rapidly enough, a
density measurements via helium pycnometry, mercury
therapeutic concentration in the body fluids may never
intrusion porosimetry, and poured and tapped bulk
be achieved, the peak plasma concentration may be
density.
significantly delayed, or much of the drug may bypass
The influence of sorbed moisture on chemical sta-
that region of the gastrointestinal (GI) tract where
bility and the flow and compaction of powders and
absorption is best. Particle size reduction (e.g., micro-
granulations is well established. The moisture content
nization) is often utilized to enhance dissolution rate.
and hygroscopicity of excipients is particularly impor-
Small particles present a larger surface area per unit
tant as total product processing as well as finished
weight to the dissolution media and hence dissolve
product stability can be affected. Hygroscopicity,
more rapidly than large particles. Particle size and
moisture-sorption isotherms, and equilibrium moisture
surface area are two of the most important properties
content can be determined by thermogravimetric
determining the solubility rate of a drug and thus
analysis and Karl Fisher titration methods.
potentially its bioavailability. There are numerous
The compactibility of relatively large-dose drug sub-
examples of bioavailability problems and bioinequiva-
stances and formulations is another important property.
lence due to the inappropriate particle size of the drug
Compactibility is of less concern for smaller-dose drugs
substance.
for which direct compression fillers may be able to com-
Particle size and shape also play an extremely impor-
pensate for a lack of ability to form mechanically strong
tant role in the homogeneity of powder blends and the
compacts. An instrumented tablet press[7] or compaction
unblending of powders in a mixer. Segregation in hand-
simulator[8] may be used to assess the relationship
ling or during the compaction process has a significant
between the mechanical strength of the compact and
effect on the content uniformity of the finished pro-
the force (or pressure) employed to form the tablet. This
ducts. Particle size can also affect the stability of a drug
relationship is the easiest of all compaction measure-
substance in that it governs the surface area available
ments to establish and provides important information
for oxidation and hydrolysis. Surface area is critical
on the ability of the material to form practical compacts.
for interaction with excipients in tablet dosage forms
Measures of compact mechanical strength include hard-
and can greatly affect stability. Methods to determine
ness (or crushing force), tensile strength, and friability.
particle size and shape include light microscopy, scan-
Other more complex studies, more easily and perhaps
ning electron microscopy, sieve analysis, and various
best done using a compaction simulator, include
electronic sensing-zone particle counters. Methods
measurement of the work or energy of compaction,
available for surface area measurement include air
pressure–density (Athy–Heckel) analysis, strain-rate
permeability and various gas adsorption techniques.
sensitivity, and elastic recovery.[9] The Athy–Heckel
analysis can provide information on deformation mech-
Bulk Powder Properties anism and give an estimate of the mean yield pressure of
the material.[10] A comparison of yield pressures determ-
Bulk powder properties are extremely important in ined at different punch speeds can give information on
pharmaceutical processing.[6] Knowledge of the true the strain-rate sensitivity of the material.[11] If the major
and bulk densities of the drug substance as well as of components of the formulation (including the drug)
the excipients is extremely useful in are strain-rate sensitive, the tablets produced on a
high-speed production press may exhibit lamination or
 Providing perspective as to the size of the final tablet capping. Excessive elastic recovery may also indicate
and the size and type of processing equipment needed, such tablet failure. The Hiestand indices (bonding and
 3644 Tablet Formulation

brittle fracture) may be used to assess the compactibility HCl, metoprolol tartrate) are expected to exhibit few
of materials under laboratory conditions.[12] bioavailability problems. On the other hand, Class II
For the evaluation of flow properties the following drugs (e.g., piroxicam) are more likely to exhibit
test methods may be used: dissolution-rate-limited absorption problems. Class III
drugs (e.g., atenolol) are more likely to be prone to
 Angle of repose absorption (permeability) rate-limited absorption.
 Minimum orifice diameter Class IV drugs (low solubility–low permeability)
 Carr index present formidable obstacles to bioavailability. An in
 Flow rate and vitro–in vivo correlation (IVIVC) is expected only in
 Direct observation of weight variation during the case of Class II drugs. An IVIVC could be expected
tableting runs. for Class I drugs if the dissolution rate is slower than
the gastric emptying the rate. With a sufficiently
The ultimate goal of flow analysis is to identify the rapidly dissolving Class I drug, little or no IVIVC is
powder or powder blend that provides the least weight expected because gastric emptying (not dissolution)
Tablet–Tablet

variation in the finished tablet. The more fluid the pow- would be the rate limiting step. Little or no IVIVC is
der is, the more efficiently and reproducibly it should fill expected for Class III or Class IV drugs.
the die cavities of a tablet press. This more efficient and The FDA has adopted the BCS in developing a
reproducible die fill should be reflected in increased tab- guidance that provides relaxed policies on scale-up
let weights and reduced intertablet weight variation.[13] and postapproval changes of immediate-release oral
solid dosage forms (SUPAC-IR). For certain changes,
requirements depend on the drug class, with the most
Solubility and Permeability liberal policies for Class I drugs, less liberal policies
for Classes II and III drugs, and the least 1iberal poli-
In many cases, the rate of dissolution in gastrointes- cies for Class IV drugs. First issued as a draft on Nov.
tinal fluids is the rate-limiting step in absorption. The bio- 29, 1994 for comment,[16,17] a revised version was
equivalence requirements established by the FDA define published in the Federal Register on Nov. 30, 1995.
low solubility as ‘‘ . . . <5 mg/mL in water, and slow The intrinsic dissolution rate (IDR) of drugs is fre-
dissolution rate to be <50% in 30 minutes.’’[14] However, quently measured in preformulation tests by the rotating
the solubility of a drug should be considered together disk method or Wood’s apparatus.[18] An automated
with its dose; that is, even a very poorly soluble drug hav- IDR system, based on a modification of a standard
ing a sufficiently small therapeutic dose may completely dissolution apparatus, allows for attachment to the stir-
dissolve under physiological conditions. Thus, Amidon rer of a die in which the pure drug has been compressed
et al.[15] have defined a ‘‘high solubility’’ drug as one with the tablet face flush with the bottom surface of the
which at the highest human dose is soluble in 250 ml die.[19] The IDR may be used to detect different poly-
(or less) water throughout the physiological pH range morphs as well as to judge the risk of a drug exhibiting
(1–8) at 37 C. A ‘‘low solubility’’ drug is thus one which dissolution-rate-limited absorption. Kaplan[20] suggested
requires more than 250 ml of water to dissolve the largest that an IDR of higher than 1 mg cm 2 min 1 indicated
human dose at any pH within the physiological range. that dissolution-related absorption problems were
The likelihood of having bioavailability problems unlikely, whereas an IDR lower than 0.1 mg cm 2 min 1
requires both a consideration of the dose and a solubility indicated dissolution-rate-limited absorption.
volume of the drug and its permeability. Amidon et al.[15]
created a Biopharmaceutics Drug Classification System
(BCS) based on estimates of these two parameters: Drug-Excipient Compatibility Studies

1. Class I: High solubility and high permeability A knowledge of the interaction of drugs and excipients
2. Class II: Low solubility and high permeability is essential in the initial formulation of a product. It
3. Class III: High solubility and low permeability may also be necessary later on during processing
4. Class IV: Low solubility and low permeability scale-up, when problems arise, to determine if incom-
patibilities exist which affect manufacturing or sta-
A jejunal permeability of at least 2–4  10 4 cm/s, bility. Drug-excipient interactions are often directly
measured in humans by an intubation technique, is related to the moisture present in one or another of
considered ‘‘high permeability.’’ For many substances, the components or to the humidity to which the formu-
this permeability corresponds to a fraction absorbed of lation is exposed during processing or storage. These
90% or better. The classification system provides a studies are always carried out at accelerated tempera-
logical basis for estimating the risk of bioavailability ture and humidity conditions, even though it must be
problems. For example, Class I drugs (e.g., propranolol recognized that some interactions are physical (melting
Tablet Formulation 3645

and volatilization) and not chemical and that acceler- strategy. Initial guidance may be provided by the
ated aging may not be predictive. Tests for excipient- proposed dose. Relatively low-dose drugs can often
drug interactions are usually conducted on blends of be tableted by direct compression, a term that is
the pure drug and excipient in ratios similar to those applied to the process by which tablets are compressed
in the final dosage form. For example, excipient- directly from blends of the active ingredient and suit-
to-drug ratios are higher for filler-binders than for lubri- able excipients. No wet or dry granulation is required,
cants and disintegrating agents. These studies are often although the drug may occasionally be sprayed out of
performed with the help of a factorial or fractional- solution onto one of the excipients to ensure uniform
factorial experimental design.[21] Powders are physically dispersion of drug in very low dosage. Larger doses
mixed and may be granulated or compacted to acceler- of poorly compactible drugs may be granulated prior
ate any possible interaction. Samples can be exposed in to tableting. The process steps required and the choice
open pans or sealed in bottles or vials to mimic product of excipients are often governed by other properties of
packaging. Evaluation of samples includes: the drug.

Tablet–Tablet
1. Visual inspection for changes in color or texture.
Analysis of Critical Variables
2. Both HPLC and TLC are commonly employed
and Formulation Development
with unstressed samples being used as controls.
In general, only qualitative results are important
Based on the analysis of the preformulation data, likely
initially.
excipients are selected and small batches may be pro-
3. Differential thermal analysis is applied and the
duced. The number and size of the batches depend
appearance or disappearance of one or more
on the availability of the drug substance. The batches
peaks is noted. Isothermal microcalorimetry
are intended to assess the feasibility of the formulation,
can also be employed as well as a thermal
including the types and levels of excipients, as well as
activity monitor (TAM) technique.
the process and its operational variables, such as order
of addition, mixing times, compression force, granu-
Compatibility studies are essential in characterizing
lation time, etc. The goal is to develop a formulation
both raw materials and finished formulations. It has
and process that meets the criteria set forth earlier
been argued that binary drug-excipient screening stu-
under Objectives.
dies are inefficient, unrealistic, and ignore processing
variables. A better approach may be to carefully select
potential excipients based on known chemistry and
MANUFACTURE
published compatibility data, and perform miniformu-
lation stability studies.[22]
Traditionally, tablets have been made by granulation,
a process that imparts two primary requisites to
Formulation Design formulations: compactibility and fluidity. Both wet
granulation and dry granulation (slugging or roll com-
Based on the preformulation information, decisions paction) are used (Table 1). Regardless of whether
can be made regarding formulation design and process tablets are made by direct compression or granulation,

Table 1 Typical unit operations involved in wet granulation, dry granulation, and direct compression
Wet granulation Dry granulation Direct compression
Milling and mixing of Milling and mixing of drugs and excipients Milling and mixing
drugs and excipients of drugs and excipients
Preparation of binder solution Compression into slugs or roll compaction Compression of tablets
Wet massing by addition of binder Milling and screening of
solution or granulating solvent slugs and compacted powder
Screening of wet mass Mixing with lubricant and disintegrant
Drying of the wet granules Compression of tablets
Screening of the dry granules
Blending with lubricants and disintegrant
to produce ‘‘running powder’’
Compression of tablets
 3646 Tablet Formulation

the first steps, milling and mixing, are the same; the addition, disintegration is optimized because directly
subsequent steps differ. compressed tablets produce primary particles upon
The wet massing of powders is typically carried out disintegration, rather than granules, which must deag-
in high-shear mixers prior to wet screening. The wet gregate to liberate primary particles. Finally, direct
granules are often dried in fluidized-bed equipment, compression tablets often exhibit fewer long-term pro-
enhancing the efficiency of the process. Alternatively, blems of chemical stability or changes in dissolution.
wet granulation may be carried out in fluid-bed drier- Although there are many significant advantages of
granulators in which the liquid phase is sprayed onto direct compression over granulation, there also are
fluidized powders while the hot air flow dries the gran- important limitations:
ules. This process reduces the number of handling steps
and the time and space needed for granulation; it can 1. Uniform blending and prevention of unblending
be automated. The advantages and disadvantages of of low-dose drugs.
wet granulation are given in Table 2. (See also Granu- 2. Fillers often are costlier than fillers used in
lations, H.G. Kristensen and T. Schaeffer, Vol. 7, 1st granulation.
Tablet–Tablet

Ed.; pp. 121–160, of this encyclopedia). 3. Physical properties and functional specifications
Regardless of the granulation method, the compara- are more critical; properties of raw materials
tive simplicity of the direct compression process offers must be defined and carefully controlled.
obvious advantages, such as 4. Limitations in producing colored tablets.
5. Dust problems.
1. Economy 6. Limitations in the dilution capacity of fillet-
2. Elimination of heat and moisture binders.
3. Optimization of tablet disintegration 7. More sensitive to lubricant softening and over-
4. Stability mixing than granulations.

The most obvious advantage of direct compression Limitations in the dilution capacity of excipients
is its greater economy, owing to reduced processing can make the direct compression of large-dose, poorly
time, less equipment and space required, less process compactible drugs impractical. Lubrication is often a
validation, and lower energy utilization. Generally, compromise between the amount and type needed for
only blending and compression are required, although adequate lubrication and their adverse effect on com-
prior micronization of the drug may be needed. Unlike pactibility. Content uniformity is of greater concern
wet granulation, processing does not require heat or in direct compression tableting, particularly with low-
moisture, which can be detrimental to drug stability. dose drugs. Since the drug is not ‘‘locked’’ into gran-
Moreover, direct compression avoids the high pres- ules, direct compression blends are subject to unmixing
sures associated with slugging or roll compaction. In in subsequent processing steps. In addition, drugs are

Table 2 Advantages and disadvantages of wet granulation

Advantages Disadvantages
Enhances fluidity and compactibility. Each unit process brings
suitable for high-dose drugs with its own set of complications
poor flow and/or compactibility
Reduces air entrapment The large number of unit
processes increases
the chances of problems
Reduces dustiness Difficult to control and validate
Provides for the addition of a Potential adverse effects
liquid phase (wet granulation) suited to of temperature, time, and
dispersion of low-dose drugs in rate of drying on drug stability
solution to ensure content uniformity and distribution during drying
Enhances wettability of powders Overall more costly than direct
through hydrophilization (wet granulation) compression in terms of space,
time, and equipment requirements
Permits handling of powders
without loss of blend quality
Tablet Formulation 3647

often micronized prior to blending to enhance their Excipients

dissolution rate, and the resulting high surface-to-mass
ratios may lead to difficulty in flowing and mixing due The design of the formulation and selection of excipients
to surface interactions. Another important limitation is is especially critical in tablet dosage forms. Products can
that unlike granulation, which tends to compensate for vary from a relatively simple aspirin tablet containing
variability in excipients, direct compression is heavily aspirin and starch to more complex systems that might
dependent upon reproducible properties of the excipi- contain fillers, binders, disintegrating agents, glidants,
ents (and the drug). Raw material standards must be lubricants, and coating agents. Modified release intro-
carefully defined and address functionality. Lot-to-lot duces even more complexity. The appropriate selection
variations in both the drug and the excipients must of excipients and their concentration are clearly critical
be avoided. The cost of raw materials and their testing to both the ability to manufacture tablets as well as to
is higher in direct compression. their performance as a drug delivery system. Since others
Thus, direct-compression tableting requires care- have illuminated the various excipient classes in great
ful attention to the choice of excipients, appropriate detail, references are provided in Table 3.

Tablet–Tablet
flow properties, and blend homogeneity, and to
the interplay of formulation and process variables Manufacturability
that can affect both compactibility and drug dissolu-
tion. (See also Direct Compression Tableting, R.F. Excipients function to provide compactibility, lubri-
Shangraw, Vol. 4, 1st Ed.; pp. 85–106, of this ency- cation, flow properties, disintegration efficiency,
clopedia.) wetting, etc. Poor choice of excipients may give rise

Table 3 Excipients literature

Excipient class References
General Rudnic, E.; Kottke, M. Tablet dosage forms.
In Modern Pharmaceutics, 3rd Ed.; Banker, G.,
Rhodes, C., Eds.; Marcel Dekker, Inc.:
New York, 1996; 333.
Diluents and fillers Czeisler, J.; Perlman, K. Encyclopedia of
Pharmaceutical Technology; Swarbrick, J.,
Boylan, J., Eds.; Marcel Dekker, Inc.:
New York, 1991; Vol. 4, 37.
Binders Kristensen, H. Encyclopedia of Pharmaceutical
Technology; Swarbrick, J., Boylan, J., Eds.;
Marcel Dekker, Inc.: New York, 1988; Vol. 1, 415.
Disintegrants Shangraw, R.; Mitrevej, A.; Shah, M. Pharm.
Tech. 1980; 4 (10), 49. Augsburger, L.L.;
Brzeczko, A.W.; Shah, U.; Hahm, H.
Encyclopedia of Pharmaceutical Technology;
Swarbrick, J., Boylan, J., Eds.; Marcel Dekker, Inc.:
New York, 2001; Vol. 20, Suppl. 3.
Lubricants and glidants Zanowiak, P. Encyclopedia of Pharmaceutical
Technology; Swarbrick, J., Boylan, J., Eds.;
Marcel Dekker, Inc.: New York, 1994; Vol. 9, 87.
Film coating agents Radebaugh, G. Encyclopedia of Pharmaceutical
Technology; Swarbrick, J., Boylan, J., Eds.;
Marcel Dekker, Inc.: New York, 1993; Vol. 6, 1.
Controlled-release agents Chien, Y. Encyclopedia of Pharmaceutical
Technology; Swarbrick, J., Boylan, J., Eds.;
Marcel Dekker, Inc.: New York, 1990; Vol. 3, 281.
Coloring agents Woznicki, E.; Schoneker, D. Encyclopedia of
Pharmaceutical Technology; Swarbrick, J., Boylan, J., Eds.;
Marcel Dekker, Inc.: New York, 1990; Vol. 3, 65.
Flavor modifiers Adjei, A., et al. Encyclopedia of Pharmaceutical
Technology; Swarbrick, J., Boylan, J., Eds.;
Marcel Dekker, Inc.: New York, 1993, Vol. 6, 101.
 3648 Tablet Formulation

to poor characteristics (hardness, appearance), which dissolution of drugs of poor water solubility formu-
can be important in packaging, storage, and patient lated into direct compression tablets.
acceptance. Problems with excipients may arise from Perhaps the greatest source of concern in excipients
variations in source or lot, particularly in formulations is with lubricants, particularly magnesium stearate,
made by direct compression. Examples of excipient which is not only hydrophobic but also has a laminar
problems include variation in performance between crystal structure. When blended with other ingredients,
Hoc cellulose and microcrystalline cellulose relative it tends to make them hydrophobic by delaminating to
to particle size, flow, and compactibility, or different coat their surfaces. The problems with magnesium
polymorphic forms of sorbitol (a, b and g) resulting stearate are thus highly process dependent. For
in tablet hardening. Differences in lactose particle size example, blending time differences of as little as
and modification (spray-dried or anhydrous) can pro- 2 min can significantly alter the dissolution pattern of
vide differences in surface areas over which a lubricant finished tablets. Because direct-compression formula-
is distributed. Anhydrous lactose hydrates at high tions have a higher specific surface area, the same
humidities with increase in size. amount of magnesium stearate blended for the same
Tablet–Tablet

length of time makes direct-compression tablet

Biopharmaceutics matrices more hydrophobic than matrices made from
granulations. The degree of shear imparted by different
The formulation of a tablet can affect its bioavailabil- mixers during processing significantly affects the distri-
ity. Particular care should be given to low-dose poorly bution of magnesium stearate.[24] The characteristics of
soluble drugs, especially those that are micronized. magnesium stearate vary from supplier to supplier and
Drugs with low water solubility should never be for- sometime within the same supplier.[25] It is essential to
mulated solely with insoluble fillers, including calcium draft raw material specifications and strictly adhere to
salts (calcium sulfate, calcium phosphate) which are Standard Operating Procedures (SOPs) during the
only soluble at very low pH. Differences in solution manufacture of products containing this lubricant.
rate between hydrated and non-hydrated forms of Establishing purchasing specifications beyond those
calcium salts as well as between dibasic and tribasic listed in the National Formulary, such as bulk and
forms may also be important. In some cases, excipients tap density, powder fluidity, particle size, surface area,
complex with drug substances such as calcium salts degree of hydration, and morphology is desirable.
with tetracycline. Varying the ratio of soluble to
insoluble fillers in tablets can significantly alter the
THE EFFECTS OF MANUFACTURING
dissolution pattern of poorly soluble drugs (weak acids
PROCESSES ON FORMULATIONS
and non-electrolytes), but has little effect on weak
bases, which are soluble in gastric fluids.
Numerous unit processes are involved in making
The type and amount of disintegrating agent can also
tablets, including particle size reduction and sizing,
be important. Differences in source (corn, potato, rice)
blending, granulating, drying, compaction, and (fre-
as well as variations in amylopectin–amylose ratios
quently) coating. Various factors associated with these
result in variable disintegration times. The fact that
processes can seriously affect content uniformity, bio-
starch included within granules is not as effective as
availability, or stability. Some of these are given in
starch added between granules has led to tablets with
the following list:
good disintegration but poor dissolution. Incorporation
of super-disintegrants (crospovidone, croscarmellose,
1. Particle Size Reduction
sodium starch glycolate) has improved dissolution from
both direct-compression and wet-granulation formula- – Non-uniform particle size can lead to segre-
tions. Both croscarmellose and sodium starch glycolate gation problems
can complex with small amounts of cationic drugs in – Development of electrostatic forces inhibits
water, but not in physiologic fluids.[23] complete blending
The type and amount of binder used in granulations – Changing the crystalline state can affect
affects dissolution rates. Many binders are hydrophilic solubility
polymers whose solubility and solubility rate depend
upon molecular weight. Quality control tests such as 2. Blending
viscosity may be necessary as part of raw material test-
ing of polymeric substances. However, an advantage of – Non-homogeneous distribution of drug sub-
the granulation process is that it results in a wetting of stance is the result of poor blending or
drug surfaces, which enhances drug dissolution once unblending
the granules have disintegrated. Wetting agents such – Overblending of lubricant lowers dissolution
as sodium lauryl sulfate can significantly improve the rates and affects compactibility
Tablet Formulation 3649

3. Granulation an ever-quickening pace to market, have prevented

the thorough examination and full understanding of
– Non-homogeneous distribution of binder most commercial formulations. Although the pace of
and drug substance gives drug-rich or formulating pharmaceutical systems has not lessened
drug-poor fines recently, a rapid turnaround means formulators must
– Decomposition of drug substance due to quickly gather information and base final decisions
residual moisture on that information. The window for error is narrower,
– Uneven granule size (too many or to few and the formulator can no longer afford to use empiri-
fines) leads to compaction or uniformity cally gathered data to obtain the critical dosage form
problems information needed for a marketable formulation.
Systematic development approaches are also desirable
4. Tableting to provide data in anticipation of the use of SUPAC
(scale up and postapproval changes) regulatory poli-
– Uneven compaction pressures affect dissolu- cies by providing for the establishment of a research

Tablet–Tablet
tion database that can help justify such changes to regulat-
– Loss of mix quality in hopper and feed ory agencies.
frame gives poor content uniformity Enormous progress has been made in the direction
– Additional shearing of lubricant in feed of systematic formulation development through the
frame lowers dissolution rates use of such statistical tools as multivariate analysis
and response surface methodology, and artificial intel-
5. Coating ligence.

– Non-uniform or incomplete coverage of

tablets and beads results in different dissolu- Experimental Design
tion patterns
In the long term, an efficient experimental design saves
When validating new equipment or procedures, the time and avoids costly mistakes, and some pharmaceu-
sampling techniques must reflect the quality of the tical firms have departments devoted to the prep-
material being tested, the blend of powders, moisture aration of experimental designs and their analysis. If
in granulation, or coating integrity. a statistician is not available, a myriad of commercial
Another potential source of problems in manu- experimental-design software packages are on the mar-
facturing is reprocessing or reworking. Reworking ket (SAS, JMP, STATGRAPHICS, DESIGN
may be required when finished products fail to meet EXPERT) with which a formulator can design experi-
hardness, content uniformity, disintegration, dis- ments on a personal computer. These same packages
solution, or appearance specifications. Reworking can also aid in data analysis and presentation.
procedures must be in writing and are often part of The early phase of such a statistical experimental
New Drug Applications (NDAs) and Abbreviated design approach may include screening designs such as
New Drug Applications (ANDAs). Although prac- two-level full or fractional factorial designs or Plackett-
ticed less today than in past years, given the high cost Burman designs. The number of experimental runs is
of some drug substances, reworking continues to be reduced by intentionally confounding some experimental
justified, but may involve the following problems: effects. Ultimately, the screening design narrows the
overdistribution of lubricant leading to poor com- number of potential variables for further study.
pactibility and dissolution; distribution of particles The latter phase of such as a central experimental
of coating in the reprocessed tablet may lower the design approach might include response surface meth-
dissolution rate; and loss in compactibility due to odologies to optimize a formulation such as a central
work hardening of direct-compression fillers, parti- composite or Box-Behnken design. Such methods
cularly at higher initial compaction pressures. allow experimenters to assess the effects of several
Reworking is less likely to cause dissolution problems variables at one time without having to study every
with water-soluble drugs. possible unique combination of variables. This way, a
systematic identification of critical variables and an
optimization of the formulation and process can be
SYSTEMATIC FORMULATION DEVELOPMENT obtained. Response surfaces are generated, which give
formulators a graphic demonstration of the effect of
Previously, pharmaceutical experimenters were rarely variables on various responses, such as drug dissolu-
afforded the luxury of using expanded experimental tion. The designs and models available to experimen-
designs. Time and manpower constraints, imposed by ters are numerous and the proper choice depends on
 3650 Tablet Formulation

the number of variables and the possible responses. immediate release formulations.[30] The development
General references for experimental design can be of a hybrid system, i.e., an Expert Network, that inte-
consulted as well as design software manuals, if a grates ANNs and ESs has the potential of taking
statistician is not available for advice. advantage of the strengths of both ANNs and ESs
and avoiding the weaknesses of either.[31,32]

Artificial Intelligence Approaches

Formulation Development
Among the artificial intelligence approaches that have
been used to provide support for the formulation Whether working in a large established laboratory or a
process are expert systems and artificial neural net- small research organization, formulators are well advised
works. An expert system (ES) is an intelligent com- to take an overall look at the development process
puter program that attempts to capture the expertise to assess the most rational approach to their particular
of specialists who have knowledge and experience in
Tablet–Tablet

needs and resources. Among many approaches to

a well-defined domain. They are designed to simulate rational tablet formulation, the strategy used by research-
an expert’s problem solving process. Developed more ers at the University of Maryland in collaboration with
than 20 years ago for other applications, expert FDA scientists[33] examines the drug product develop-
systems are a relatively new idea in pharmaceutical ment process in light of the SUPAC guidance.
technology. In rule-based systems, the knowledge is This generalized plan focuses on the assessment the
highly structured and often represented as a set of rules possible influence of formulation and processing vari-
that express the relationship between several pieces of ables on bioavailability and manufacture. The research
information in the form of conditional statements. protocol is best visualized by the flow chart in Fig. 1.
These statements specify actions to be taken or advice The model begins with the preformulation stage
to be followed. Such systems can shorten development wherein information is obtained on the physicochemical
time, simplify formulations, provide the rationale for and biopharmaceutic properties of the drug. One out-
decisions taken in arriving at a formulation, serve as come of the preformulation study should be the identi-
excellent teaching tools for novices, and accumulate fication of the biopharmaceutic class (BCS) in which the
and preserve the knowledge and experience of experts. drug falls since this will provide important guidance in
ESs suffer from the limitation that they are not cre- making formulation decisions. For multisourced drugs,
ative. They can only deal with situations that have the different sources are considered. The preformulation
been anticipated. They must be designed to handle
every contingency. Examples include expert systems
developed for formulating tablets,[26] for process trou- Biopharmaceutic/
Preformulation
bleshooting,[27] and for the selection of a mixer.[28] Pharmacokinetic Profile
Used in other disciplines for about 40 years, arti- Physicochemical
ficial neural networks (ANNs), like ESs, have only Multiple Characterization
recently been applied to pharmaceutical development. Drug
ANNs are computer-based programs that attempt to Sources Critical
Experimental Design
Variables
simulate certain functions of the biological brain, such Analysis In Process Analysis
as learning, generalizing, or abstracting from experi- Dissolution
ence. They have the ability to discern relationships or Response Surfaces
Formulation Selection
patterns in response to exposure to facts (‘‘learning.’’).
ANN models may be viewed simply as multiple non-
SUPAC
linear regression models. Through ANNs, the data Small Scale
Recommended Dissolution Profiles
and information generated during experimental work Ranges Manufacture
may be transformed relatively easily into knowledge
that would enable the formulator to at least construct
a few domain specific rules, even though confined, for
Bioavailability Study
future cases. However, ANN effectiveness is limited by Biostudy In Vivo - In Vitro
the training data selected. One limitation is that, in Relationship
most cases, ANNs lack explanation capability and
there is difficulty in obtaining justification for results.
Examples of applications to product development Scale-Up
include predicting model granulation and tablet
characteristics from knowledge of material and pro- Fig. 1 A research model used in a University of Maryland
cess variables,[29] and predicting drug release from and FDA collaborative research program. (From Ref.[33].)
Tablet Formulation 3651

study results in a portfolio of information, which 2. Czeisler, J.L.; Penman, K.P.; Diluents. Encyclopedia of Phar-
provides guidance in formulation design and in the maceutical Technology, 1st Ed.; Swarbrick, J., Boylan, J.C.,
Eds.; Marcel Dekker, Inc.: New York, 1991; Vol. 4, 40–43.
development of appropriate hypotheses to be tested 3. Blecher, L.; Ohmae, T.; deJong, H.J. Tri-PEC: reports from
and the critical variables studies that follow. IPEC-Americas, JPEC, and IPEC-Europe. Pharm. Tech.
In the critical variables analysis phase, a statistical 1994, 18 (8), 53.
4. Raleblian, J.; McCrone, W.J. Pharmaceutical applications
experimental design is created (e.g., factorial, Box– of polymorphism. Pharm. Sci. 1969, 58, 911.
Behnken) intended to assess critical formulation and 5. Matsumoto, T.; Nobuyoshi, K.; Iliguchi, S.; Otsuka, M.J.
process variables in relatively small-scale manufacture. Effect of temperature and pressure during compression on
polymorphic transformation and crushing strength of
In these studies, the ranges of composition variables chlorpropamide tablets. Pharm. Pharmacol. 1991, 43, 74.
are chosen to at least encompass those noted in the 6. Brittain, H.G. Raw materials. Drug Devel. Ind. Pharm.
recommendations of the AAPS–FDA Workshop on 1989, 15, 2083.
7. Schwatz, J.B. The instrumented tablet press: uses in
Scale-up of Immediate Release Oral Solid Dosage research and production. Pharm. Tech. 1981, 5 (9), 102.
Forms or SUPAC. This phase is usually preceded by 8. Hunter, B.M.; Fisher, D.G.; Pratt, R.W.; Rowe, R.C. A
a development phase during which variables and levels high speed compression simulator. J. Pharm. Pharmacol.

Tablet–Tablet
1976, 28, 65.
to be studied are determined and the exact method of 9. Maganti, L.; Celik, M. Compaction studies on pellets I.
manufacture is established. Experimental formulations uncoated pellets. Int. J. Pharm. 1993, 95, 29.
are assessed at least in terms of dissolution perform- 10. Muller, F.X.; Augsburger, L.L. The role of the displace-
ment-time waveform in the determination of heckel beha-
ance, content uniformity, and weight variation. On vior under dynamic conditions in a compaction simulator
the basis of these studies, the specific formulations to and a fully instrumented rotary tablet machine. J. Pharm.
be manufactured for biostudy are selected. Pharmacol. 1994, 46, 468.
11. Roberts, R.J.; Rowe, R.C. The effect of punch-velocity on
In the small-scale clinical manufacturing phase, for- the compaction of a variety of materials. J. Pharm. Pharma-
mulations are manufactured under GMP conditions col. 1985, 37, 377.
for possible clinical testing. These will be manufactured 12. Hiestand, E.; Smith, D.P. Indices of tableting performance.
Powder Tech. 1984, 38, 145.
on a larger scale than those in the previous phases. 13. Augsburger, L.L.; Shangraw, R.F. Effect of glidants in
Anexperimental design is chosen; however, if some of tableting. J. Pharm. Sci 1966, 55, 418.
the variables can be eliminated based on the earlier 14. Fed. Reg. In 21CFR, Ch. 1; (4/1/87 Ed.) Part 32; 320.
15. Amidon, G.L.; Lennernas, H.; Shah, V.P.; Crison, J.R. A
experiments, the number of formulations produced theoretical basis for a biopharmaceutic drug classification:
may be reduced. the correlation of in vitro drug product dissolution and in
The intent of the biostudy phase is to establish an vivo bioavailability. Pharm. Res. 1995, 12, 413.
16. INTGUIDE.1R9. In FDA Center for Drug Evaluation and
in vitro–in vivo relationship. If an appropriate corre- Research; Rockville, MD, 1994.
lation can be established, dissolution may serve as a 17. Lucisano, L.J.; Franz, R.M. FDA proposed guidance for
surrogate for biostudies in the interpretation of what is chemistry, manufacturing, and controls changes for
immediate-release solid dosage forms: a review and indus-
significant and what is not among the variables studied. trial perspective. Pharm. Tech. 1995, 19 (5).
In the scale-up phase, larger runs of formulations 18. Wood, J.H.; Syarto, J.E.; Letterman, H.J. Improved holder
are manufactured. The formulations are selected to for intrinsic dissolution rate studies. Pharm. Sci. 1965, 54,
1068.
determine if the larger scale will enhance the signifi- 19. Koparkar, A.D.; Augsburger, L.L.; Shangraw, R.F. Intrin-
cance of certain variables. sic dissolution rates of tablet filler-binders and their influ-
The statistical analysis of the data provides the ence on the dissolution of drugs from tablet formulations.
Pharm. Res. 1990, 7, 80.
opportunity to predict changes in dissolution perform- 20. Kaplan, S.A. Biopharmaceutical considerations in drug
ance resulting from incremental changes in one or formulation design and evaluation. Drug Metab. Rev.
more formulation or process variable at a time (e.g., 1972, 1, 15.
21. Jacobs, A.L. Determining optimum drug/excipient com-
the level of an excipient or the time of mixing). patibility through preformulation testing. Pharm. Manuf.
The development model presented here may differ 1985, 2 (6), 43.
from some formulation research programs in that bio- 22. Monkhouse, D.C.; Maderich, A. Whither compatibility
testing. Drug Devel. Ind. Pharm. 1989, 15, 2115.
studies may not be performed on small-scale batches. 23. Hollenbeck, R.G.; Mitrevej, K.; Fan, A. Estimation of the
The major advantage of early biostudies is the poten- extent of drug-excipient interactions involving croscarmel-
tial for early IVIVC and subsequent surrogate use of lose sodium. Pharm. Sci. 1983, 72, 325.
24. Bolhuis, G.K.; deJong, S.W.; Lerk, C.F. The effect of mag-
dissolution testing in further work. Scale-up and nesium stearate admixing in different types of laboratory
pilot-plant roles in formulation changes, while not and industrial mixers on tablet crushing strength. Drug
covered thoroughly here, are reviewed by Racz.[34] Devel. Ind. Pharm. 1987, 13, 1547.
25. Danserean, R.; Peck, G.E. The effect of the variability in
the physical and chemical properties of magnesium stearate
on the properties of compressed tablets. Drug Devel. Ind.
Pharm. 1987, 13, 975.
REFERENCES 26. Rowe, R.C. An expert system for the formulation of tablets.
DTI Manuf. Intel. Newsletr. 1993, 14, 13–15.
1. Chowhan, Z.T. Excipients and their functionality in drug 27. Murray, F.J. The application of expert systems to pharma-
product development. Pharm. Tech. 1993, 17 (9), 72. ceutical processing. Pharm. Tech. 1989, 13 (3), 100–110.
 3652 Tablet Formulation

28. Lai, F.K.Y. A prototype expert system for selecting BIBLIOGRAPHY

pharmaceutical powder mixers. Pharm Tech. 1988, 12 (8),
22–31.
American Pharmaceutical Association and the Royal Pharma-
29. Kesavan, J.G.; Peck, G. Pharmaceutical granulation and
tablet formulation using neural networks. Pharm. Dev. ceutical Society of Great Britain. In Handbook of Pharma-
Tech. 1996, 1 (4), 391–404. ceutical Excipients, 3rd Ed.; American Pharmaceutical
30. Ebube, N.K.; McCall, T.; Chen, Y.; Meyer, M.C. Relating Association and Pharmaceutical Press: Washington, 2000.
formulation variables to in vitro dissolution using an Armstrong, A.M.; James, K.C. Understanding Experimental
artificial neural network. Pharm. Dev. Tech. 1997, 2 (3), Design and Interpretation in Pharmaceutics; Ellis Horwood
225–232. Ltd: Chichester, UK, 1990.
31. Turban, E. Expert Systems and Applied Artificial Intelli- Franz, R.M.; Browne, J.; Lewis, A. Experimental design, model-
gence; Macmillan Publishing Co.: New York, 1992. ing, and optimization strategies for product and process
32. Caudill, M. Expert networks. In Neural Network PC Tools; development. In Pharmaceutical Dosage Forms: Disperse
Academic Press: New York, 1990; 189–214. Systems, 2nd Ed.; Lieberman, H.L., Rieger, M., Banker, G.,
33. Augsburger, L.L.; Shangraw, R.; Lesko, L.; Williams, R. Eds.; Marcel Dekker, Inc.: New York, 1996; 1, 427–514.
An approach toward establishing a scientific foundation Peck, G.E.; McCurdy, V.E.; Banker, G.S. Tablet formulation
for interpreting regulations and workshop reports on and design. In Pharmaceutical Dosage Forms: Tablets, 2nd
scale-up and post approval changes. Pharm. Res. 1994, Ed.; Lieberman, H.L., Lachman, L., Schwartz, J., Eds.;
Tablet–Tablet

11 (10), S-161. Marcel Dekker, Inc.: New York, 1989; 1, 75–130.

34. Racz, I. Drug Formulation; John Wiley & Sons: NewYork, Polderman, J. Formulation and Preparation of Dosage Forms;
1989; 25–27. Elsevier/North-Holland, New York, 1977; 3–28.
Tablet Manufacture
Norman Anthony Armstrong
Welsh School of Pharmacy, Cardiff University, Cardiff, U.K.

INTRODUCTION of tablets on a commercial scale, and this led to the

concentration of pharmaceutical manufacture in rela-
The compressed tablet is by far the most widely used tively few industrial sites.
dosage form, having advantages for both producer A monograph for Glyceryl Trinitrate Tablets was
and user. However, the manufacture of tablets can be included in the British Pharmacopoeia of 1885, but

Tablet–Tablet
a complex process, since only a few raw materials no other tablet monograph appeared there until
inherently possess those properties which are necessary 1945. This was not due to lack of popularity of the dos-
for the production of tablets of satisfactory quality. age form itself, but rather the absence of suitable meth-
Hence some preliminary treatment and/or incor- ods of quality control that were applicable to tablets.
poration of excipients in the formulation is usually The tablet did not meet with universal approval. In
needed. Tablet manufacture is a paradox. Considerable 1895 an editorial in the Pharmaceutical Journal in the
ingenuity and formulation expertise are required to United Kingdom described the tablet as ‘‘one of the
transform a mass of particles into a low porosity mass. evils suffered by legitimate pharmacy,’’ and predicted
Yet, after the tablet has been ingested, the requirement that tablets ‘‘have had their day.’’[1] Not withstanding
then is usually for the tablet to release its active such a prediction, the usage of tablets has continued to
ingredient as rapidly as possible, and further ingenuity increase. The 2000 edition of the British Pharmaco-
is needed to bring this about. poeia contains 320 monographs for tablets, far in
Tablets are solid preparations each of which con- excess of any other dosage form.
tains a single dose of one or more active ingredients. The tablet is the most popular dosage form because
They are obtained by compressing uniform volumes it provides advantages for all concerned in the pro-
of particles, and are almost always intended for oral duction and consumption of medicinal products.
administration. Though the initial capital outlay for the manufacturer
The earliest reference to a dosage form resembling of tablets is considerable, they can be produced at a
the tablet is to be found in tenth century Arabic medi- much higher rate than any other dosage form, tablet
cal literature. Drug particles were compressed between presses capable of producing about one million tablets
the ends of engraved ebony rods, force being applied per hour being available. Furthermore, the fact that
by means of a hammer. Details of the tabletting pro- the tablet is a dry dosage form promotes stability,
cess, as it is now known, were first published in 1843 and in general, tablets have shelf lives measured in
when William Brockedon was granted British Patent years. They are also convenient to transport in bulk,
9977 for ‘‘manufacturing pills and medicinal lozenges since they contain relatively small proportions of exci-
by causing materials when in a state of granulation, pients unlike, for example, oral liquids.
dust or powder, to be made into form and solidified From the viewpoint of the pharmacist, tablets are
by pressure in dies.’’ In this case, too, force was applied easy to dispense, while the patient receives a concen-
by a hammer. Potassium bicarbonate was the first trated and readily transportable and consumed dosage
pharmaceutical substance to be so treated. form. Furthermore, if properly prepared, tablets pro-
The use of compressed pills, as they were then vide a uniformity of dosage greater than that of most
known, increased rapidly. It is likely that the term other medicines, and appropriate coating can mask
‘‘tablet’’ for this dosage form was first used in the unpleasant tastes and improve patient acceptance.
United States in the 1870s. Power-driven presses The tablet also provides a versatile drug delivery
replaced Brockedon’s hammer, and by 1874 there system. Though most tablets are intended to be swal-
existed both rotary and excentric presses, which in lowed intact, the same basic manufacturing process,
their mode of operation were fundamentally similar to associated with appropriate formulation, provides
those in use at the present time. The tablet lent itself medicines for sublingual, buccal, rectal, and vaginal
to mass manufacture by mechanical means, in contrast administration, together with lozenges, soluble, dis-
to the slower labour-intensive production of older solid persible, and effervescent tablets. In addition, techni-
dosage forms such as the pill. It is impractical for ques that can delay or otherwise modify the release
individual pharmacists to produce small quantities of the active ingredient from the tablet are available.
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-100001726
Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved. 3653
 3654 Tablet Manufacture

Naturally tablets only possess these advantages if of gravity from a hopper. Though tablets are usually
they are properly formulated and manufactured. A well- described in terms of weight, the die is filled by a volu-
prepared tablet should possess the following qualities: metric process. The volume is determined by the depth
to which the lower punch descends in the die. Unless
1. It should, within permitted limits, contain the this volume is filled reproducibly on each occasion,
stated dose of drug. then the mass of the tablet will vary, and with it the
2. It should be sufficiently strong to withstand the drug content of each tablet. Therefore, uniform filling
stresses of manufacture, transport, and handling is essential. However, it must be borne in mind that
so as to reach the patient intact. the die cavity has a cross-section of only a few milli-
3. It should deliver its dose of drug at the site and metres, and only a fraction of a second is available
at the speed required. for filling each die. It therefore follows that the parti-
4. Its size, taste, and appearance should not cles must flow easily and reproducibly.
detract from its acceptability by the patient.
Tablet–Tablet

Stage 2. Compression
TABLET COMPRESSION The upper punch descends, and its tip enters the
die, confining the particles. The distance separating
All tablets are made by a process of compression. the punch faces decreases, either by movement of the
Solid, in the form of relatively small particles, is con- upper punch alone (as in excentric presses) or by move-
tained in a die and a compressing force of several ment of both punches (as happens in rotary presses).
tonnes is applied to it by means of punches. The shape The porosity of the contents of the die is progressively
of the die governs the cross-sectional shape of the tab- reduced, and the particles are forced into ever-closer
let, and the distance between the punch tips at the proximity to each other. This process is facilitated by
point of maximum compression governs its thickness. the particles fragmenting and/or deforming. Once the
The conformation of the tablet faces, usually flat or particles are close enough together, interparticulate
convex, is a reflection of those of the punches. forces then cause the individual particles to aggregate,
The tip of the lower punch moves up and down forming a tablet. The magnitude of the force is gov-
within the die, but never actually leaves it. The upper erned by the minimum distance separating the punch
punch descends to penetrate the die and apply the faces. Therefore, a second essential property of the par-
compressive force. It is then withdrawn to permit ejec- ticles is that they cohere under the influence of a com-
tion of the tablet, brought about by an upward move- pressive force. It is also essential that this coherence be
ment of the lower punch. maintained when the compressing force is removed.
There are two types of tablet press. The excentric
press has one die and one pair of punches. The rotary
press has a larger number of dies which are fitted, with Stage 3. Ejection
their corresponding punches, into a rotating turret.
Irrespective of the type of press that is used, the pro- The upper punch is withdrawn from the die, and so the
cess of tablet compression can be divided into three force being applied to the tablet is removed. The effect
stages, as shown in Fig. 1. of this might be to cause the deformed particles to
return to their former shape, which would result in a
decrease in interparticulate contact and hence tablet
Stage 1. Filling strength. It is essential that this does not occur. As
the upper punch leaves the die, the lower punch moves
The lower punch falls within the die, leaving a cavity upwards, pushing the tablet before it. During the com-
into which particulate matter flows under the influence pression stage, the particles are forced into intimate
contact with the interior die wall. It follows that
attempts to remove the tablet will be opposed by fric-
Upper punch
Tablet tional forces and so successful ejection demands lack of
adhesion between the tablet and the diewall.
Die Therefore in summary, for a particulate solid to be
successfully transformed into tablets, three key proper-
Lower punch ties need to be present:
Ejection

Filling Compression
1. Good particle flow.
2. The ability of the particles to cohere under the
Fig. 1 Cycle of operations of an excentric tablet press. influence of a compressing force. This coherence
Tablet Manufacture 3655

must be retained after the compressing force has Active ingredient(s) Mixing Diluent
been removed.
3. The ability of the tablet to be ejected from the die Binder Wetting Water
after the compressing force has been removed.
Granulation
Few powders possess all these essentials and some
possess none of them. Thus, before successful tablet-
Drying
ting can take place, some preliminary treatment with
the addition of one or more excipients is almost
invariably needed. Sieving

Glidant Mixing
lubricant Disintegrating agent
METHODS OF TABLET PRODUCTION

Tablet–Tablet
Compression
The pretreatment that is usually necessary takes the
form of granulation. The process of granulation is Fig. 2 The wet granulation process of tablet manufacture.
essentially one of size enlargement, and it serves several
purposes in the tablet manufacturing process:
necessary to increase the bulk of such a tablet with a
diluent. Some commonly used diluents are listed in
1. It improves flow by increasing particle size,
Table 1.
since large particles flow more readily than
The ideal diluent would be both chemically and
small ones.
physiologically inert, and would not interfere with
2. It improves compression characteristics, adding
the bioavailability of the active ingredient. It should
to the cohesive strength of the tablet.
also be inexpensive and be easily tabletted since, if the
3. Once a homogeneous mixture has been achieved,
proportion of active ingredient is small, the overall tablet-
segregation is prevented, since particles that are
ting properties of the mixture are largely governed by
stuck together cannot separate.
those of the diluent.
4. It reduces dust.
Lactose is by far the most frequently used diluent
for solid dosage forms. An inexpensive disaccharide
Both wet and dry granulation techniques are
obtained as a by-product of the cheese industry; it is
available.
available in a number of forms, though a-Lactose
monohydrate is the variety that is normally used as
Tablet Manufacture by Wet Granulation the diluent in tablets made by wet granulation. It is
freely albeit slowly soluble in water and as such it is
This is the traditional method of pretreatment of solids a suitable diluent for active ingredients of low water
prior to tabletting. Despite its complexity and inherent solubility. Lactose is a non-reducing sugar, and is
disadvantages, even now about half the tablets pro- reasonably inert. It can take part in the Maillard reac-
duced worldwide are manufactured by this process. tion when mixed with substances containing primary
Its essence is that particles of active ingredient, with amine groups, giving highly colored products, and thus
a diluent if necessary, are stuck together using an its use is contraindicated in such formulations.[2]
adhesive, the latter usually being water-based. The Probably the second most commonly used diluent
result is a granular product which flows more readily in the wet granulation process is dibasic calcium
and has an improved ability to cohere during com- phosphate. This substance is virtually insoluble in
pression. water and hence is always used in conjunction with a
A flow diagram of the wet granulation process disintegrating agent. Its properties have been reviewed
together with appropriate excipients is shown in Fig. 2. by Carstensen and Ertell.[3]

The diluent Mixing

The first stage in the wet granulation process is often a The purpose of the mixing stage is to ensure that the
dry mixing stage in which the active component is powder blend and hence the resulting tablets are
mixed with a diluent. Many drugs need to be adminis- homogeneous in content. A random mixture is defined
tered in doses of only a few milligrams or even less, yet as one where the probability of sampling a given type
a tablet that weighs less than about 50 mg is difficult of particle is proportional to the number of such parti-
for the patient to handle conveniently. It is therefore cles in the total mixture. Thus, the aim is to produce a
 3656 Tablet Manufacture

Table 1 Tablet diluents

Diluent Comments
Calcium carbonate Insoluble in water (Cal-Carb , MillicarbÕ, Pharma-CarbÕ, SturcalÕ)
Õ

Calcium phosphate, dibasic Insoluble in water, good flow properties, available in dihydrate
and anhydrous forms (CyfosÕ, CalstarÕ, CalipharmÕ, EmcompressÕ)
Calcium phosphate, tribasic Insoluble in water (TricafosÕ, Tri-CalÕ, Tri-TabÕ)
Calcium sulfate Insoluble in water (Cal-TabÕ, CompactrolÕ)
Cellulose, microcrystalline Good compression properties, may not need lubricant, can act as
disintegrant (AvicelÕ, EmcocelÕ, VivacelÕ)
Cellulose, microcrystalline silicified Combination of microcrystalline cellulose and silica (ProsolvÕ)
Cellulose, powdered (ElcemaÕ, Solka-FlocÕ)
Dextrates (EmdexÕ)
Tablet–Tablet

Dextrose Hygroscopic, reducing sugar (TabfineÕ)

Fructose (FructofinÕ)
Lactitol (FinlacÕ)
Lactose monohydrate The most commonly used diluent. Inexpensive, takes part in Maillard
reaction (Fast-FloÕ, LactochemÕ, MicrotoseÕ, PharmatoseÕ,
TablettoseÕ, ZeparoxÕ)
Magnesium carbonate
Maltitol (MaltisorbÕ, MaltitÕ)
Maltodextrin (GlycidexÕ, LycatabÕ, MaltrinÕ)
Maltose (AdvantoseÕ)
Mannitol Freely soluble in water, negative heat of solution and therefore
cool taste, popular for chewable tablets, non-cariogenic (PearlitolÕ)
Sodium chloride Freely soluble in water, used in solution tablets
Sorbitol
Starch Also acts as disintegrating agent, may give soft tablets
Starch, pregelatinized Also acts as disintegrating agent. (LycatabÕ, Pharma-GelÕ, Pre-JelÕ,
SepistabÕ, Starch 1500Õ, Starx 1500Õ)
Sucrose Freely soluble in water, sweet taste, hygroscopic, used in lozenges in
conjunction with lactose
Sugar, compressible (DipacÕ, NutabÕ)
Sugar, confectioner’s
Sugar spheres (Nu-CoreÕ, Nu-PareilÕ)
Talc
Xylitol Negative heat of solution, cool taste (XylifinÕ, XylitabÕ)
Proprietary names are given in parentheses.

mixture such that when a sample is removed, the this is not always the case. Under certain conditions,
relative proportions of the components of that sample an optimum mixing time occurs, beyond which the
are the same as in the mixture as a whole. mixture shows a tendency to separate back into its
Unlike molecules in a fluid, which in time will mix components. This process is known as segregation.
spontaneously by a diffusion mechanism, powder par- Segregation is particularly likely to occur in mixtures
ticles do not mix spontaneously but remain in their where the components differ markedly in size, with dif-
relative positions. Therefore before mixing can occur, ferences in shape and density as secondary factors. It is
energy must be put into the system. This causes the especially likely to occur if regular patterns of move-
powder bed to dilate or expand, the particles separate ment are set up in the mixing device, and for this
from one another and this leads to relative motion reason, mixers are designed so that an irregular mixing
among them. motion occurs.[4]
It might be intuitively expected that the randomness Although in general a size difference between com-
of a mixture will progressively increase with time, but ponents can lead to segregation, a situation where
Tablet Manufacture 3657

there is a large difference in sizes between components can be established. Ertell et al. have shown that the size
may be beneficial. In such circumstances, small parti- of the granulator and the mixing time can be major
cles of one component can become trapped in irregula- influences on the physical properties and dissolution
rities in the surface of the larger component. These are rate of the resulting tablets.[7]
not random mixtures, as the particles of the two com- As described above, the wet granulation process is a
ponents cannot behave independently. This concept is long and hence expensive procedure, which has been
called ‘‘ordered mixing’’ and it has found applicability improved by the introduction of high-speed mixer
in the manufacture of solid dosage forms containing granulators. These have agitator and chopping blades,
small quantities of highly potent active ingredients[5] which enable mixing, wet massing, and granulation to
(see the article on Blenders and Blending in this encyc- take place in the same piece of apparatus. In such
lopedia). devices, the granulation process takes place extremely
rapidly, and hence the establishment of optimum
Granulation granulation times is even more important.
A further technique is fluid-bed granulation. Air is

Tablet–Tablet
The underlying process of size enlargement in wet passed into the powder bed from below. This causes
granulation is achieved by either one or both of two the particle, of powder to form a suspension in the
different mechanisms. Firstly, adjacent solid particles air and gives effective mixing. The granulating fluid
may be stuck together using an adhesive. Such sub- is then sprayed over the particles, which adhere on col-
stances are known as binders or granulating agents. lision and they are then dried in the heated air stream.
Secondly, dissolution of the solid in the granulating The wet granulation process, apparatus, and phar-
liquid can occur, followed by evaporation of the liquid maceutical applications have been comprehensively
phase of the latter. This will result in the deposition reviewed by Kristensen and Schaefer[8] (see the article
of dissolved material on particle surfaces, forming on Tablet Granulation in this encyclopedia).
so-called crystal bridges. The occurrence of this mech-
anism will depend on the solubility of the solids in the Drying
liquid phase. Thus, sucrose will form crystal bridges
with an aqueous granulating fluid, whereas calcium After the process of granulation, the product exists as a
phosphate will not. wet mass from which the liquid must be removed, since
The process and underlying mechanisms of granu- the presence of water leads to the impairment of flow
lation have been fully described by Sherrington and properties, and perhaps to chemical instability. Water
Oliver.[6] Details of commonly used binders are given is usually removed by evaporation for which energy
in Table 2. They are often natural or synthetic poly- is needed. This is normally provided as heat, though
mers and are usually added as aqueous solutions or microwave energy is being increasingly used for drying
dispersions. Alternatively, they can be mixed with the in tablet manufacture.
other solids in the formulation in the dry state, water The essential constituents of an effective piece of
then being added. drying equipment are a heat supply to increase the
If the active ingredient is unstable in the presence of temperature and thereby reduce relative humidity, a
water, then a granulation process using non-aqueous device for removal of evaporated water and a means
liquids can be used. The usual granulating system in of minimizing the distance that water molecules must
such cases is povidone dissolved in isopropanol. The diffuse before they can be evaporated.
extra costs and environmental problems posed by the The fluidized bed drier is the most commonly used
use of a volatile and flammable liquid are disincentives device for drying tablet granules. The solid is fluidized
to the use of non-aqueous granulation. from below by a jet of hot air, and so each granule
The traditional piece of granulating apparatus is the becomes separated from its neighbors. The air provides
shear granulator. Its function is to homogeneously an effective means of heat transfer, as well as of remov-
incorporate an adhesive and viscous liquid such as ing water vapor. The speed of the drying process is
starch paste into a mass of dry powder to form governed by the distance that water molecules must
agglomerates. It follows that a considerable shearing diffuse before they arrive at the evaporative surface.
force needs to be exerted. The mixed solids are loaded Since the wet granules are present as individual units,
into the bowl of the mixer, and the liquid added with the maximum distance over which diffusion occurs is
agitation. The damp solid is then forced through a equal to the radius of a granule. Hence, fluidized bed
relatively coarse screen (about 1–2 mm), often by means drying is a rapid process.
of oscillating bars, to give discrete granules. The pro- The temperature of the bed can be precisely con-
gression of the granulation process can be monitored trolled, and a free-flowing product results. The resem-
by measuring the electrical power consumption by blance to fluid-bed granulation will be apparent,
the granulator, and hence optimum granulation times and apparatus based on the fluidized bed principle is
 3658 Tablet Manufacture

Table 2 Binders used in the wet granulation process

Concentration in the
Binder granulating fluid (% w/v) Comments
Acacia mucilage Up to 20 Yields very hard granules
Alginic acid 1–5
Carbomer 5–10 (CarbopolÕ)
Carboxymethylcellulose calcium 5–15 (NymcelÕ)
Carboxymethylcellulose sodium 5–15 (NymcelÕ)
Cellulose, microcrystalline (AvicelÕ, EmcocelÕ, VivacelÕ)
Powdered cellulose (ElcemaÕ, Solka FlocÕ)
Ethyl cellulose 1–3 (AquacoatÕ)
Gelatin 5–20 Forms gel in cold water, therefore
Tablet–Tablet

warm solution used, strong adhesive

Glucose, liquid Up to 50 Strong adhesive, hygroscopic
Guar gum 1–10
Hydroxyethyl cellulose 2–6 (CellosizeÕ)
Hydroxypropyl cellulose 2–6 (KlucelÕ, MethocelÕ)
Hydroxypropyl cellulose—low-substituted 5–25
Hydroxypropylmethyl cellulose 2–5 (MethocelÕ, PharmacoatÕ)
Magnesium aluminum silicate 2–10 (PharmasorbÕ, VeegumÕ)
Maltodextrin 2–10 (GlucidexÕ, LycatabÕ, MaltrinÕ)
Methylcellulose 1–5 (CelacolÕ, MethocelÕ)
Polydextrose (LitesseÕ)
Polyethylene oxide 5 (PolyoxÕ)
Povidone 0.5–5 Also known as PVP or polyvinylpyrrolidone.
Soluble in water and some organic solvents, can
be used for non-aqueous granulation, very
commonly used, synthetic material (KollidonÕ,
PlasdoneÕ)
Sodium alginate 1–3 (ManucolÕ)
Starch paste 5–25 Very commonly used
Starch, pregelatinized 5–10 (LycatabÕ, Pharma-GelÕ, Pre-JelÕ, SepistabÕ,
Starch 1500Õ, Starx 1500Õ)
Sucrose (syrup) Up to 70 Hygroscopic, tablets may harden on storage
Water Suitable for solids that are freely soluble in water
Proprietary names are given in parentheses.

available in which mixing, granulation, and drying may have to diffuse through the whole thickness of
take place in the same chamber. the solid layer before the evaporative surface is
Although the apparent turbulence of the air stream reached. Thinner layers give quicker evaporation, but
may give rise to interparticulate collisions and hence this would reduce the overall capacity of the drier.
attrition, this is not usually a severe problem. However, Thus, the drying process is slower in a tray drier than
the rapid movement of particles in a hot, dry atmos- in a fluidized bed drier.
phere can lead to the development of static electrical As water diffuses through the bed of solid, it will
discharges. Suitable precautions must therefore be carry with it any components of the formulation that
taken, especially if flammable liquids have been used are soluble in it. This will lead to a non-uniform distri-
in the granulation process. bution of these components in the solid. This is not
A more traditional means of drying is the tray drier. usually a problem with fluidized bed drying, but with
Hot air flows over a series of shelves on which the wet tray drying, significant differences in composition can
material is spread. Compared to the fluidized bed drier, occur between the upper and lower surfaces of the
the solid–air interface is smaller, and water molecules solid bed. This can give rise to non-uniform drug
Tablet Manufacture 3659

content and, if the migrating species is colored, Methods of assessing glidant action have been
variation in the appearance of the product.[9,10] reviewed by Augsburger and Shangraw.[11] Lerk et al.
Microwaves are being increasingly employed in the showed that a concentration of 0.2% colloidal silica
pharmaceutical industry for drying purposes. The inci- in a tablet formulation had no effect on tablet crushing
dent microwave radiation (frequencies of 2450 and strength. However, higher concentrations reduced
960 MHz are used) causes electrons in substances such crushing strength especially when associated with pro-
as water to resonate, which in turn generates heat and longed mixing times.[12] Some commonly used glidants
causes the water to evaporate. The water vapor is are shown in Table 3.
removed under vacuum, and hence the product dries
rapidly at a relatively low temperature. As the bed of
solid is stationary, particle attrition does not occur, The lubricant
and dust formation is minimized.
When the tablet formulation is compressed, the sides
of the tablet are brought into intimate contact with

Tablet–Tablet
Second mixing stage the die wall. The tablet must then be ejected from the
die, involving the movement of the side of the tablet
When the drying process is complete, it is likely that relative to the die wall. Therefore, friction between
the product will have cohered into relatively large the tablet and the die wall must be overcome. With
masses, especially if tray drying has been used. The materials such as lactose, friction resistance can be
dried material is therefore passed through a sieve considerable, and it may be impossible to remove the
(usually 250–700 mm) to break up aggregates and to tablet from the die without damage to the tablet or
give a relatively uniformly sized granule. A second to the tablet press. Therefore, a lubricant is almost
mixing stage now follows in which several important invariably included in a tablet formulation. A lubricant
ingredients of the formulation are added. is a substance that deforms easily when sheared
between two surfaces, and hence when interposed
The glidant between the tablet and the die wall, provides a readily
deformable film.[13] Details of some tablet lubricants
The formation of granules from the original powder are shown in Table 4.
particles may have improved flow sufficiently for uni- Inadequate lubrication can often be recognized by
form die filling to be achieved. However, if flow is still vertical scratches on the sides of the tablet. It may also
not good enough, a glidant (also known as an anticak- lead to a build-up of solid on the punch faces, which in
ing agent) can be added to improve flow still further. turn gives a matt, dimpled appearance to the face of
The most frequently used glidant is colloidal silicon the tablet, a phenomenon known as picking.
dioxide, which has a mean size of about 20 nm. It is In practice, magnesium stearate is by far the most
thought to act by lodging in the surface irregularities frequently used tablet lubricant, and is extremely effec-
of the granule, forming a more rounded structure tive. Its activity, as with other metallic salts of fatty
and hence reducing interparticulate friction. Colloidal acids, is believed to derive from adhesion of the polar
silica has the added advantage of acting as a moisture metallic portion of the molecule to the powder particle
scavenger. Residual water in the formulation is bound surface. As a consequence, the hydrocarbon portion
to the silica, thereby providing a drier environment for of the molecule becomes oriented away from the
the other ingredients. surface.[14] Thus, a non-polar layer is presented to

Table 3 Tablet glidants

Glidant Concentration in tablet (%) Comments
Calcium silicate 0.5–2
Cellulose, powdered 1–2 (ElcemaÕ, Solka FlocÕ)
Magnesium carbonate 1–3
Magnesium oxide 1–3
Magnesium silicate 0.5–2
Silicon dioxide, colloidal 0.05–0.5 Excellent glidant (AerosilÕ, Cab-o-SilÕ)
Starch 2–10
Talc 1–10 Insoluble in water but not hydrophobic
Proprietary names are given in parentheses.
 3660 Tablet Manufacture

Table 4 Tablet lubricants

Lubricant Concentration in tablet (wt%) Comments
Calcium stearate 0.5–2 Water insoluble
Fumaric acid 5 Water soluble
Glyceryl behenate 0.5–4 Water insoluble
Glyceryl palmitostearate 0.5–5.0 Water insoluble (PrecirolÕ)
Hydrogenated vegetable oil 1–6 Water insoluble, may be used in conjunction
with talc (LubritabÕ, SterotexÕ)
Magnesium lauryl sulfate 1–2 Soluble in warm water
Magnesium stearate 0.25–5 Water insoluble, excellent lubricant, reduces
tablet strength, prolongs disintegration
and dissolution times
Tablet–Tablet

Polyethylene glycol 4000 or 6000 2–5 Soluble in water, moderately effective, also
known as macrogols (CarbowaxÕ)
Sodium lauryl sulfate 1–2 Water soluble, moderate lubricant, but good
wetting properties, often employed in
conjunction with stearates (EmpicolÕ, Stearowet CÕ)
Sodium stearyl fumarate 0.5–2.0 Sparingly soluble in cold water, soluble in
hot water (PruvÕ)
Starch 2–10 Moderate lubricant
Stearic acid 1–3 Water insoluble
Talc 1–10 Insoluble in water but not hydrophobic.
A moderate lubricant
Zinc stearate 0.5–2 Water insoluble
Proprietary names are given in parentheses.

adjacent powder particles and structures such as the uncontaminated by lubricant, is created as the particles
press tooling. It is from the formation of this non-polar break up. This new surface can then take part in inter-
layer that the advantages and disadvantages of the use particulate bonding.[15]
of magnesium stearate in a tabletarise. All these factors, both positive and negative, are
To act as an effective lubricant in a tablet, the lubri- consequences of the attrition of particles of lubricant
cant must be dispersed over the surface of the powder and their spreading around the exterior surface of the
particles or granules. The more complete this layer, the other components of the tablet. Therefore, any proces-
more effective the lubricant action will be. However, sing factor that can affect lubricant attrition or the
this has two deleterious consequences. The first is that completeness of the film might be expected to influence
each powder particle presents a hydrophobic and tablet disintegration, dissolution, bioavailability, and
hence water repellent exterior. It is well known that physical strength. The mixing process is extremely
the presence of a lubricant based on fatty acids slows important here, and mixing time, mixer type, and batch
disintegration and dissolution, and has been shown size[16] have all been shown to influence tablet proper-
to cause bioavailability problems. ties. Thus, there is a need to establish a minimum lubri-
The second consequence is that direct contact cant concentration and an optimum mixing time
between powder particles is, at least in part, replaced within which adequate lubrication is achieved without
by contact between adjacent hydrocarbon layers. Since the development of undesirable tablet characteristics.
these by definition have low shear strength, it is not To ensure batch-to-batch uniformity, the parameters
surprising that interparticulate bonding is reduced of the mixing process such as type of mixer, batch size,
and hence the tablet structure is weakened. Reduction and mixing time must be kept as constant as possible.
in tablet strength is particularly marked with sub- A mixing time of 2–5 min usually suffices to give
stances such as microcrystalline cellulose that undergo adequate lubrication.[17]
deformation on compression, since although the parti- The water repellent properties of hydrocarbon
cles may change shape, the hydrocarbon layer remains based lubricants can be countered to a certain extent
intact. Substances which fragment on compression suf- by the inclusion of a wetting agent such as sodium
fer a smaller reduction in strength, since new surface, lauryl sulfate into the formulation. Such materials
Tablet Manufacture 3661

themselves can have a limited lubricant action. Mix- only broke down the tablet but also the constituent
tures of stearates and lauryl sulfates are commercially granules, giving a finer product.[21]
available. The mechanism of action of disintegrating agents
Sodium stearyl fumarate has been used as an alter- has been the subject of some debate.[22] Some sub-
native for magnesium stearate. It has about the same stances such as starch swell when they come into con-
lubricating effect, and causes similar tablet strength tact with water, and disruption of the tablet structure
reduction and prolongation of disintegration time.[18] has been attributed to this. However, other effective
Lubricants based on fatty acids, because of their low disintegrants do not swell in this way, and are believed
water solubility, are unsuitable for tablets which must to act by providing a network of hydrophilic pathways
be dissolved in water before use. Polyethylene glycol inside the tablet through which water can diffuse. Irres-
6000 (macrogol 6000) is soluble in water, but its lubri- pective of the precise mechanism of disintegration, it is
cant activity is limited. Magnesium lauryl sulfate has clear that water uptake into the tablet must be the first
been suggested as a water-soluble substitute for mag- step in the disintegration process.[23]
nesium stearate. In addition to its lubricant action, this Addition of wetting agents such as sodium lauryl

Tablet–Tablet
substance, like sodium lauryl sulfate, is an effective sulfate or sodium docusate can assist this water
wetting agent.[19] penetration by lowering the surface tension, and
It must be stressed that the functions of a glidant they are often used in conjunction with hydrophobic
and lubricant in a tablet formulation are totally differ- lubricants such as magnesium stearate (see the article
ent. A few materials, e.g. talc, can act as both glidant on Tablet Disintegrants and Disintegration in this
and lubricant, but usually two different excipients are encyclopedia).
needed. Thus, although colloidal silicon dioxide is an
excellent glidant, it has no lubricant activity. Con-
versely, magnesium stearate, despite its popularity as Tablet Manufacture by Dry Granulation
a lubricant, can hinder rather than promote flow.
Although widely used, the wet granulation method of
tablet manufacture suffers from several disadvantages.
The disintegrating agent Water is the usual granulating fluid, and this exposes
tablet ingredients to the danger of hydrolysis. Further-
Strongly coherent particles are essential for the pro- more, the granulating fluid has to be removed, usually
duction of robust tablets, which will have high physical by heating. In addition to the energy costs that are
strength and low porosity. However, before it can be incurred, the elevated temperature will accelerate any
absorbed in the gastrointestinal tract, the active hydrolytic reaction that might be taking place.
ingredient must dissolve, and a physically strong tablet Dry granulation is an alternative method that can
is an impediment to dissolution. Therefore, tablet for- be used, and this process is shown in Fig. 3. The com-
mulations often include a disintegrating agent, which ponents of the formulation are compressed in the dry
when it comes into contact with water, disrupts the state. If sufficient bonding strength cannot be achieved
tablet structure and leads to fragmentation. A larger by compression alone, a binder is added, also in the
surface area is thus exposed to the dissolving fluid dry state.
and dissolution is facilitated. Tablets which contain a The initial compression stage can take place by one
large proportion of solids that are freely soluble in of two methods. The first uses a conventional tablet
water have less need of a disintegrating agent, since press, a process often referred to as ‘‘slugging.’’
such tablets tend to erode from their exterior surfaces Because the components of the formulation will not
rather than disintegrate. Details of some tablet disinte- have the necessary attributes for producing good
grating agents are given in Table 5. tablets, the tablets produced at this stage (the slugs)
For many years, starch was the disintegrating agent will not be of acceptable quality, especially as regards
of choice. Recently, however, so-called ‘‘super disinte- to appearance and weight uniformity. The slugs are
grants’’ have been introduced, which markedly reduce then broken down to form a granular product, which
tablet disintegration time. Such substances include after sieving can then be compressed again to give sat-
croscarmellose, crospovidone, polacrilin potassium, isfactory tablets. Malkowska and Khan showed that
and sodium starch glycolate.[20] the ease of compressibility of the formulation at the
The disintegrating agent may be mixed with other second compression was inversely proportional to the
powders prior to wetting with the granulating fluid pressure used at the slugging stage, implying that slug-
(intragranular) or at the second mixing stage (extragra- ging at high pressure should be avoided.[24]
nular), or both. Shotton and Leonard found that while A second method of compression is to use a roller
extragranular disintegrating agents caused the tablet to compactor. The powder mixture is passed between
disintegrate quicker, intragranular disintegrants not two contra-rotating cylindrical rollers to form a cake,
 3662 Tablet Manufacture

Table 5 Tablet disintegrating agents

Disintegrating agent Concentration in tablet (wt%) Comments
Alginic acid 2–10
Carbon dioxide Created in situ in effervescent tablets
Carboxymethylcellulose calcium 1–15 (NymcelÕ)
Carboxymethylcellulose sodium 1–5 (NymcelÕ)
Cellulose, microcrystalline Up to 10 Directly compressible, some lubricant
properties (AvicelÕ, EmcocelÕ, VivacelÕ)
Cellulose, powdered 5–15 Solka FlocÕ
Croscarmellose sodium 0.5–5 (Ac–di-SolÕ, SolutabÕ)
Crospovidone 2–5 (Kollidon CLÕ, Polyplasdone XLÕ)
Docusate sodium 0.5–1 Acts primarily as a wetting agent
Tablet–Tablet

Guar gum 2–8

Hydroxypropyl cellulose—low-substituted 5–25
Magnesium aluminum silicate 2–10 (VeegumÕ)
Methylcellulose 2–10
Polacrilin potassium 2–10 Cation exchange resin
(Amberlite IRP88Õ)
Poloxamer 5–10
Povidone 0.5–5 (KollidonÕ, PlasdoneÕ)
Sodium alginate 2.5–10 (ManucolÕ)
Sodium glycine carbonate Source of carbon dioxide for
effervescent tablets
Sodium lauryl sulfate 0.5–2 Primarily a wetting agent but
aids disintegration (EmpicolÕ)
Sodium starch glycolate 2–8 (ExplotabÕ, PrimojelÕ)
Starch 2–10 Potato and maize starches are
most frequently used
Starch, pregelatinized 5–10 (LycatabÕ, Pharma-GelÕ, Pre-JelÕ,
SepistabÕ, Starch 1500Õ, Starx 1500Õ)
Proprietary names are given in parentheses.

which as before is broken down to a product of granu- The key component here is the diluent. This must
lar size and then recompressed. Both methods require not only possess those properties which are necessary
the addition of a lubricant prior to the first com- for satisfactory tablet formulation, but also retain
pression stage, though more lubricant will probably those properties when mixed with the other constitu-
be needed before the second compression. ents of the formulation such as the active ingredient.
The process of direct compression is shown in
Fig. 4. The ingredients are mixed together and then
Tablet Manufacture by compressed. Almost invariably a lubricant must be
Direct Compression added, and a glidant and a disintegrating agent
included when necessary. The process does not involve
Both wet and dry granulation methods of tablet the use of a liquid, and hence a drying stage with its
manufacture are complex multistage processes, but attendant energy costs is avoided.
are necessary to convert the components of the formu- Details of some direct compression diluents are
lation into a state that can be readily compressed into given in Table 6. The majority of these are available
acceptable tablets. If, however, a major component of from only one supplier, though the two most fre-
the formulation already possesses the necessary degree quently used—spray-dried lactose and microcrystalline
of fluidity and compressibility, granulation would be cellulose—are available from several sources.
unnecessary. This is the basis of the direct compression In view of the apparent simplicity of this method
method of tablet manufacture.[25] of tablet manufacture and the number of suitable
Tablet Manufacture 3663

Active ingredient(s) Diluent The true direct compression process as described

earlier almost invariably applies to formulations con-
(Binder) Lubricant
taining potent active ingredients and where the direct
Mixing
compression properties derive from the diluent. A
few substances do possess adequate flow and cohesive
First compression properties without the need for pretreatment. These
(by tablet press or roller compactor) are usually crystalline inorganic salts such as sodium
chloride and potassium chloride. Direct compression
forms of less potent active ingredients are available
Comminution
e.g., paracetamol and ascorbic acid. These can be
directly compressed into tablets, perhaps after the
Sieving
addition of a lubricant. However, such substances are
Glidant more accurately described as ‘‘pre-granulated,’’ in that
Mixing Disintegrating agent
lubricant the granulation process—either wet granulation or

Tablet–Tablet
precompression—has been carried out by the excipient
Second compression manufacturer.
Fig. 3 The dry granulation process of tablet manufacture.

THE BEHAVIOR OF PARTICLES UNDER

diluents that are commercially available, it is perhaps A COMPRESSIVE LOAD
surprising that techniques of tablet manufacture
involving granulation are still so widely used. Direct All tablet manufacture can be regarded as the appli-
compression can, of course, only be used when a diluent cation of pressure to a population of particles enclosed
is required by the formulation, i.e., the active ingredient in a confined space. An understanding of particle
must be relatively potent. Direct compression can offer behavior under such conditions is therefore the key to
significant savings in energy, equipment, and material understanding the formation and properties of tablets.
handling costs. Against this must be set higher ingredi-
ent costs, since direct compression diluents are more
Application of a Force to Particles in a Die
expensive than other diluents.
There are, however, other factors which must be
Attractive forces exist between any two solid bodies.
considered. In wet granulation, the properties of the
These forces may be non-specific, e.g., van der Waal’s
individual drug and diluent particles are, at least to a
forces, or may be more specific in nature, e.g., brought
certain extent, hidden by the binder, whereas in direct
about by molecules exhibiting intermolecular hydrogen
compression, the original particles are still present.
bonds. However, irrespective of their nature, it is these
Therefore, in the latter technique, a premium is placed
forces acting among a large population of particles
on batch-to-batch consistency of particulate properties
that enable a coherent tablet to be formed.[26] Their
such as size and shape for both drug and diluent. In a
magnitude depends directly on the particle mass and
direct compression formulation, the components can
inversely on the square of the distance separating
behave as individual particles, and therefore there is
the particles. It follows, therefore, that with small
a danger that these can segregate after mixing and
particles of small mass, a tablet will only be formed
prior to compression. In a granulation process, the
when adjacent particles are forced into intimate
particles are bound together and so segregation is less
contact with each other. This contact is brought about
likely to happen. Furthermore, the reduction in dust
by the application of force.
formation brought about by granulation cannot occur
A representation of what may happen to an indi-
in direct compression.
vidual particle when a force is applied to it can be
obtained by considering what happens to a spring
when subjected to a load that is applied and then
Active ingredient(s) Diluent removed. This is shown in Fig. 5. Although the ana-
logy of a powder under compression to a spring under-
going elongation is not exact, it does provide useful
Glidant Mixing Lubricant
comparisons. The load is termed the stress and the
change in length the strain.
Compression
Disintegrating agent Initially there is a rectilinear relationship between
stress and strain, and if the stress is removed, the spring
Fig. 4 The direct compression process of tablet manufacture. returns to its original length. This is elastic behavior
 3664 Tablet Manufacture

Table 6 Direct compression tablet diluents

Diluent Proprietary name Comments
Õ Õ
Calcium phosphate, dibasic Emcompress , Di-Tab Good flow properties, high density,
insoluble in water
Calcium phosphate, tribasic Tri-TabÕ Insoluble in water
Calcium sulfate CompactrolÕ Insoluble in water
Õ Õ Õ
Cellulose, microcrystalline Avicel , Emcocel , Vivacel Highly compressible, low bulk density,
acts as disintegrant
Cellulose, powdered ElcemaÕ
Dextrates EmdexÕ
Lactitol FinlacÕ DC
Lactose
Tablet–Tablet

Anhydrous alpha Pharmatose DCL30Õ Good flow properties

Anhydrous beta Pharmatose DCL21Õ
Spray-dried Fast-FloÕ, ZeparoxÕ,
Pharmatose DCL11Õ
Lactose-cellulose coprocessed mixture CellactoseÕ
Maltodextrin LycatabÕ, MaltrinÕ Fairly soluble in water,
slight lubricant effect
Mannitol PearlitolÕ Freely soluble in water,
negative heat of solution
Sorbitol NeosorbÕ
Starch, pregelatinized starch Starch 1500Õ, Starx 1500Õ Disintegrant
Õ Õ Õ
Sucrose–maltodextrin coprecipitate Des-Tab , Dipac , Nu-Tab Good flow properties,
moisture sensitive
Xylitol XylitabÕ Freely soluble in water,
negative heat of solution

and is completely reversible. The spring is said to obey until eventually the load is so great that the breaking
Hooke’s law and the reciprocal of the slope of this por- point of the spring is reached and it snaps. Now, con-
tion of the curve is Young’s Modulus for the spring. sider now a number of particles constrained in the die
If the stress is further increased, eventually a point is of a tablet press and to which a progressively increasing
reached when the straight-line relationship is lost. This force is applied. A series of events can then occur, per-
is termed the elastic limit. If stresses in excess of the haps sequentially but there is a greater likelihood that
elastic limit are applied and then removed, the spring some overlap will occur.
will not return to its original length. Thus, a fraction The particles will undergo rearrangement to form a
of the change in length is permanent or irreversible, less porous structure. This will take place at very low
and this is termed plastic behavior. Further increase forces, the particles sliding past each other. This stage
in load will result in more and more plastic deformation will usually be associated with some fragmentation, as
the rough surfaces move relatively to one another and
asperities are abraded away.
The particles have now reached the stage where
Breaking point relative movement becomes impossible, although the
porosity of the powder bed may still be considerable.
A further increase in applied force can then induce
elastic deformation, plastic deformation, or fragmen-
Strain Elastic limit tation. Which of these alternatives predominates will
Slope = (Young's modulus−1) depend on the properties of the material involved,
Permanent
deformation
Hookean deformation but the net result will be a further decrease in porosity,
and an increase in interparticulate contact.
Stress
If only elastic deformation has occurred, then when
Fig. 5 Stress–strain relationships. the compressing force is removed, the particles will
Tablet Manufacture 3665

return to their former shape. The additional inter- much lower in the powder mass on its vertical axis.
particulate contact caused by compression will be lost On the other hand, low force zones occur on the same
and a coherent tablet will not be formed Fig. 6. axis but much nearer to the moving punch Fig. 7.
If, however, the elastic limit has been passed, then as Train’s findings were later confirmed by Charlton
the force is removed, not all the increased interparticu- and Newton using gamma-ray attenuation.[28]
late contact will be lost, cohesion will be retained and a The consequences of such a force distribution on
tablet will be formed. Thus, from the point of view of tablet strength can be profound. Particle deformation,
forming a robust tablet, substances with low elastic whether elastic or plastic, will be proportional to the
limits, which undergo plastic deformation at low com- force applied, and as has been discussed, this defor-
pressive forces, are preferable to more elastic bodies. mation is an essential preliminary to the formation of
If consolidation of the powder mass is brought the interparticulate bonds on which tablet integrity
about by fragmentation, then a large number of points depends. Thus, the porosity of the tablet, and hence
of interparticulate contact are created, from which the its strength, will vary within the tablet. The weakest
strength of the tablet derives. In this case, removal of points in the tablet structure will be those that receive

Tablet–Tablet
the compressing force should have no effect on tablet the lowest force i.e., on the face of the tablet
strength, since there is no way the fragments can adjacent to the stationary punch and on the central
recombine into the original particles. However, purely axis near to the moving punch. Thus, because of its
fragmentary consolidation is unlikely to have occurred, non-uniform density, some parts of a tablet are stron-
and so the effect of removal of the force on deformed ger than others.
particles must still be considered. It should be noted that this discussion assumes that
only one punch is actively applying the force to the
powder mass while the other is stationary and passive.
Force Transmission through a Powder Bed This is true in the case of an excentric press, but with a
rotary tablet press, both punches move and hence both
Consider as before a group of particles in the die of an exert forces on the powder bed. The force distribution
excentric tablet press. Force is applied by means of the so obtained is thus different from that shown in Fig. 7,
descending upper punch and because the lower punch and results in two low density zones near the faces of
is passive, the force will be transmitted to it through the tablet and a high density zone in approximately
the powder bed. The distribution of force within the the centre of the powder mass.
powder bed was investigated by Train, who embedded The effect of the removal of the compressing force
force transducers (q.v.) in a relatively large mass of must now be considered. Elastic recovery will occur
powder.[27] He found that the diminution of force did to a greater or lesser extent, which will result in a
not proceed uniformly on descent through the bed, reduction in the strength of interparticulate bonds
but formed a much more complex pattern. This was and an overall weakening of the tablet. It therefore
caused by the forces being transmitted to and reflected follows that if a tablet is to be disrupted by elastic
from the die wall. Significant features are zones of high recovery, this is most likely to occur at its weakest
force at the periphery near the moving punch, and point. This is just below the top surface, and is the
phenomenon often encountered in tablet manufacture
known as lamination or capping. With this explanation
in mind, some effects associated with capping, and some
causes and pragmatic solutions to the problem can now
be explained.
Compressive load applied Capping was for many years considered to be due
to the entrapment of air in the tablet, and even the
Particles Particles
deform fragment
under under
load load
Compressive load removed

Low density High density

zones zones

Plastic deformation Elastic deformation Particles

Particles remain deformed Particles return to former shape remain
Cohesion maintained Cohesion lost fragmented
Fig. 7 Density distribution in a compact prepared on an
Fig. 6 Elasticity and plasticity in a particulate mass. excentric tablet press.
 3666 Tablet Manufacture

production of tablets in vacuo which still capped did time, dissolution time are dependent in some way on
little to dispel this theory. Neither did this suggestion the force that is applied by the punches to the particles
explain why air should cause the fracture just below in the die.
the face of the tablet. However, by considering the Considerable research on tablet properties was
non-uniform density distribution in the tablet, it can performed for many years, but until a method of
be seen that the weakness is not caused by the presence accurately measuring compression force was available,
of air per se, but rather the relative absence of solid in meaningful studies could not be carried out.
those parts of the tablet that have high porosity.[29] As The key to progress in this field was the introduc-
compression proceeds, it follows that the pores in the tion of the so-called instrumented tablet press by
tablet structure are filled with air at a progressively ele- Higuchi and others in the mid 1950s,[31] in which force
vated pressure, and this will obviously assist disruption transducers were fitted to the press to measure the
of the tablet when the compressing force is removed. applied load. This revolutionized research into the
Thus, any factor which obstructs the expulsion of air tabletting process and in addition has lead to the devel-
from the powder mass during compression will exacer- opment of presses with automatic tablet weight
Tablet–Tablet

bate capping, though it is not the fundamental cause. control, since the mass of particles in the die governs
Such factors include the clearance between punch the force detected by the transducer[32] (see the article
and die, the speed at which the force is applied, and on Automation of Tablet Presses in this encyclopedia).
the presence of small particles, which makes passage A force transducer is also known as a strain gauge.
of air through the tablet more tortuous.[30] In its simplest form, the strain gauge is a network of
Any applied stress that exceeds the breaking wires through which an electric current is passed. The
strength of the tablet will also cause the tablet to break wires are bonded very securely to a component of the
at its weakest point. A number of stresses occur when press, e.g., the upper punch. If a force is applied to
the tablet is removed from the die after compression. the punch, it deforms. The magnitude of the defor-
The die may become worn at the point in the die where mation (the strain) is governed by a combination of
the tablet is compressed, i.e., the die is fractionally the magnitude of the force (the stress) and the value
wider at this point than elsewhere. Thus, when the of Young’s modulus for the material from which the
tablet is ejected, it is forced through an aperture, the punch is made. The wire of the strain gauge is also
diameter of which is slightly less than that of the tablet deformed, and hence its electrical resistance changes.
itself. This will obviously stress the tablet, and the If the gauge is incorporated into a Wheatstone bridge
interparticulate bonds may be overcome at their weak- circuit, then a small change in voltage results. The size
est point. Also as the tablet is extruded from the die, of the signal from the strain gauge is proportional to
elastic expansion will occur not just in an axial but in the amount of deformation, which in turn is a function
a radial direction. The latter occurs progressively, i.e., of the applied force. Hence, after amplification and
one segment of tablet is free to expand while the one appropriate calibration, the voltage changes can be
below is still constrained by the die. Bond disruption expressed in terms of the applied force. Signals from
will be an inevitable consequence. the transducers can be fed into an oscilloscope or chart
recorder, or stored electronically and subsequently
manipulated by computer.
CHARACTERIZATION OF THE A further advance was the fitting of displacement
COMPACTION PROCESS transducers to tablet presses. These too give out an
electrical signal, the magnitude of which is governed
A range of parameters has been devised which by the position of a sensing device in relation to a fixed
attempts to describe the process of powder compac- reference point, and in this way punch movement can
tion, both to elucidate principles and also to enable be measured. As before, these signals can be amplified,
predictions to be made regarding compaction proper- recorded, and stored. If the outputs of the force and
ties. The majority of these depend on the availability displacement transducers are combined, the applied
of methods of accurately measuring applied force force at a given point in time, and punch position at
and punch position. the same instant can be established.
Instrumentation of rotary presses is more difficult
than that for an excentric press if transducers are to
Measurement of Applied Force be fitted directly to the punches. Since the turret of
and Punch Movement the press rotates, a fixed connection between trans-
ducers and the recording device is impracticable.
The aim of any tabletting process is to produce tablets Alternatives to fixed connections are slip rings and
that are of satisfactory quality. Virtually all tablet radiotelemetry. A further option is to fit transducers
properties e.g., porosity, physical strength, disintegration to points of the press remote from the punches and
Tablet Manufacture 3667

which do not rotate. Techniques for fitting transducers punch does not fall to zero until ejection is complete
to both excentric and rotary tablet presses, their (point J). The greater the ejection force, the greater
calibration, and applications of the information the need for a lubricant in the formulation.
derived from instrumented presses have been reviewed The reason why the lower punch maximum force is
by Watt.[33] less than that of the upper punch is because a fraction
Fig. 8 shows the output from force and displace- of the force applied by the upper punch is transmitted
ment transducers fitted to both punches of an excentric to the die wall, where it results in die wall friction. This
tablet press. The upper punch describes an approxi- is reduced by the presence of a lubricant. Hence, the
mately sinusoidal path as it descends to penetrate the ratio between lower punch maximum force and upper
die (point A) and then rises after the compression event punch maximum force, often called the R value, is used
has taken place, leaving the die at point B. The lower as the basis of comparison between lubricants.[34] R has
punch remains motionless during the compression a maximum value of unity and lubricants based on
event and then rises to eject the tablet from the die stearates usually exhibit R values greater than 0.95.
(point C). The time that elapses from point D to point F is

Tablet–Tablet
As the upper punch enters the die and comes into known as the consolidation time. This is the time inter-
contact with the particles, the height of the bed of par- val when a force is detectable at the upper punch. The
ticles is reduced and hence porosity decreases. Initially contact time is the period when the upper punch is in
porosity reduction is brought about by particle contact with the original particles or the tablet (point
rearrangement. This requires a very low force which D to point H). The residence time is the period when
is probably not detectable by the force transducers, a force is detected at the lower punch (point D to point
the output of which remains zero. The upper punch J), which ends on tablet ejection.
then encounters a resistance to its motion (point D) Transducer output from a rotary tablet press differs
as further consolidation by rearrangement becomes in two aspects. Firstly, the lower punch plays an active
impossible. Hence, the output of the upper punch force role in the compression event and moves upwards as
transducer rises, slowly at first and then more rapidly. the upper punch moves downwards. The second differ-
Particles are deformed and/or fragmented during this ence is small but important. Because of the sinusoidal
stage to form a coherent tablet. Force is transmitted movement of the upper punch in an excentric press,
to the lower punch, and a similar rise is detected by the punch speed is only zero at the instant when it
transducers there. As maximum upper punch pen- reverses direction (point E). Punches on a rotary press
etration is achieved (point E), force maxima are have a flat area on the punch head. As the punches
detected on both punches (points F and G), that on pass under or over the pressure rolls, the flat area
the lower punch being less than that on the upper. dictates that there is no punch movement. The period
Once the maxima have occurred, the upper punch during which this occurs is called the dwell time, and
begins to rise, and the force detected on both punches though it only lasts a fraction of a second, it can have
falls. That on the upper punch returns to zero as a major effect on the consolidation process.[35,36]
contact is lost between the ascending punch and the
top surface of the tablet (point H). That on the lower
Tablet Strength Profiles

The physical strength of a tablet is dependent on the

Upper punch A B extent and strength of interparticulate bonds and these
movement E in turn are related to the compressive force which
is applied. Therefore, the relationship between the
Lower punch C
applied force and some parameter related to tablet
movement strength is a good indication of the ease with which
a given substance will form satisfactory tablets, and
F
Upper punch may also give an insight into the compaction mech-
force G
anism of the solid and its mechanical properties.
The strength is usually assessed as the force required
Lower punch to fracture a tablet in a defined direction e.g., its diam-
force eter. This has traditionally been referred to as the
‘‘hardness’’ of the tablet, which is incorrect termi-
D H J nology in this context. In material science, hardness
Time
is a surface property related to resistance to inden-
Fig. 8 Force and displacement data from an instrumented tation. Terms such as physical strength or mechanical
excentric tablet press. strength are more appropriate.
 3668 Tablet Manufacture

The compression force is measured using an instru- Confusion can often arise in the units that are used
mented tablet press as described earlier. The physical to express compression force or pressure and the
strength of the tablet is measured with crushing strength of the tablet. Table 7 gives examples of units
apparatus such as that described in most current phar- that have been used recently for the axes of tablet
macopoeias. The results are conventionally presented strength profiles. Comparison between different tablet
graphically with compression force as the abscissa formulations would be greatly facilitated if authors
and strength as the ordinate. used and journal editors insisted on the use of the SI
This technique is satisfactory when comparing system of units. The SI unit of length is the meter, that
tablets of the same size and shape, such as in-process of force the Newton and that of pressure the Pascal.
control. However, if virtually identical tablets are not The unit for physical strength is the Newton and that
being tested, problems may arise. The force is some- of tensile strength calculated from Eq. (1) is the Pascal.
times expressed as the compression pressure, obtained
by dividing the force by the cross-sectional area of the
punch. This is valid if the punch has a flat surface, the Relationships between Tablet Porosity and
Tablet–Tablet

area of which can be easily calculated; but in practice Compression Force or Pressure
this is often not the case, and conversion between
compression force and compression pressure involves Fig. 8 shows the movement of both the upper and
assumptions regarding the area of the punch face, lower punches of the tablet press in relation to fixed
which may not be valid. reference points and from these data, the distance
The physical strength of a tablet is also dependent separating the two punch faces can be calculated. If
on its dimensions. In the construction of a force- both punch faces are in contact with the tablet, it
strength profile, all tablets will have the same cross- follows that the distance of their separation is equal
sectional area as the same tooling will have been used. to the height of the tablet (h). Consequently, if the
However, as the compressive force is changed, so will cross-sectional area of the die (a), the tablet weight (w)
the tablet height. Hence, comparisons made on the and the true density of the solid from which the tablet
basis of breaking strength will not be truly valid. is made (r) are all known, the porosity of the tablet (Z)
This problem has been circumvented in part by the can be calculated from Eq. (2).
calculation of the tensile strength of the tablet. The
most commonly used formula is shown in Eq. (1),
introduced by Rudnick, Hunter, and Holden[37] and e ¼ 1
ðw=ahrÞ ð2Þ
Fell and Newton.[38]
For cylindrical tablets, Eq. (2) becomes Eq. (3)
Ts ¼ 2P=Pdt ð1Þ e ¼ 1
ð4w=Pd2 hrÞ ð3Þ

where Ts is the tensile strength of the tablet (MPa), P As the applied force (or pressure) being applied at
the crushing strength (N), d the tablet diameter (m), the same moment in time can also be determined from
and t the tablet thickness (m). Fig. 8, it follows that a relationship between force and
This equation applies to cylindrical tablets which porosity can be constructed.
have a diameter and whose height is constant over A typical relationship is shown in Fig. 9. As force
the whole tablet surface. Newton et al. have attempted is increased from zero, the porosity of the tablet
to extend the concept of tensile strength to tablets that falls rapidly, but then further increase in force has a
are not cylindrical.[39] progressively smaller effect. The porosity value at

Table 7 Some units used for the construction of tablet strength profiles
Abscissa parameter Unit Ordinate parameter Unit
Force kg Crushing strength (hardness) kg
lb Strong-Cobb units
kN N
N kp
Pressure Kg cm
2 Tensile strength Kg cm
2
Pa Pa
MPa MPa
lbin
2
Tablet Manufacture 3669

ln 1/(1– D)
Porosity

Pressure Pressure

Tablet–Tablet
Fig. 9 A force-porosity diagram. Fig. 10 Pressure and porosity data plotted according to the
Heckel equation.

which the curve becomes virtually horizontal is depen-

dent on the solid being compressed. Substances that tablet presses were not widely available. The height
deform plastically typically give tablets of lower of the tablet was measured after ejection from the die
porosity than those which fragment. and thus the net change in height was a combination
Many equations have been derived in attempts to of height reduction by consolidation and a height
provide a mathematical expression of Fig. 9. These increase by elastic expansion. Furthermore, the typical
have been reviewed by Kawakita and Ludde.[40] It plot comprised only a small number of points, and an
must be stressed that such equations are simply math- apparently rectilinear zone could be identified. With
ematical descriptions of the data, and they have no improvements in instrumentation and in particular
underlying physical significance. linkage to a computer, Heckel plots of several hundred
The most widely used of these equations is the points became feasible with tablet height measure-
Heckel relationship:[41] ments being made before ejection from the die. In the
author’s experience, the more points there are on a
pressure–porosity diagram, the more difficult it is to
ln 1=ð1
DÞ ¼ kP þ A ð4Þ identify a truly rectilinear section.
Paronen and Ilkka have surveyed the use of the
where D is the relative density of the tablet and hence Heckel and other compression equations and have
(1
D) is the porosity, P is the applied pressure, and drawn attention to difficulties in their interpretation.[43]
k and A are constants.
This equation predicts that a graph of ln 1/(1
D)
against P should yield a straight line of slope k and Force–Displacement Curves
intercept A. Heckel surmised that the greater the slope,
the greater degree of plasticity in the solid being com- Fig. 8 shows how force and punch displacement
pressed. Hersey and Rees[42] defined the reciprocal of k change with time. It is therefore possible to determine
to be the mean yield pressure of the solid. both the force and displacement values at any given
Fig. 10 shows a typical Heckel diagram. Deviations point in time, and from this, a force–displacement
from linearity often occur at low and high pressures. curve can be constructed. Usually the force is plotted
These are to be expected. At low pressures, reduction as the ordinate and displacement as the abscissa
in porosity is largely by particle rearrangement, and Fig. 11.
thus the true consolidation mechanism, i.e., fragmen- The area enclosed by the curve has the units of force
tation or deformation, will be a minor component of (Newtons) multiplied by distance (metres), which is
the total consolidation process. At high pressures, dimensionally equivalent to work or energy (in Joules).
porosity can become very low and hence its reciprocal Therefore, the force–displacement curve has been used
becomes a very large number. to calculate the work consumed in the compression of
However, the real problem with Heckel plots is a solid.[44]
identifying a truly rectilinear section. When pressure– Further refinement of this technique has enabled
porosity diagrams were first devised, instrumented estimates of elasticity and plasticity of the solid to be
 3670 Tablet Manufacture

B Time-Dependent Effects

It has long been known that for some formulations,

changing the speed of operation of the tablet press or
changing the type of press can have a profound effect
Force on tablet quality. Such circumstances may arise, for
example, when changing from a slow excentric press
in formulation development to a high speed rotary in
production.
As described earlier, diagrams of transducer output
against time enable parameters such as contact time
and dwell time to be defined Fig. 8. Also the slope of
A D C the displacement–time diagram equals the speed of the
Displacement
punch and the slope of the force–time diagram is the
Tablet–Tablet

Fig. 11 A force-displacement curve. rate of change of the force. If the operating speed
of the press is altered, there is a proportional change
in all of these.
Such considerations are important because the
made. If the particles were totally non-elastic, then as consolidation of many substances is time-dependent.
soon as the upper punch reached its maximum pen- Fragmentation can be regarded as a virtually instan-
etration and began to rise again, contact would be lost taneous process. However, substances that undergo
with the tablet. Therefore, the force recorded by trans- deformation behave in a viscoelastic manner, and the
ducers fitted to the upper punch would instantly fall to time over which the compression force is applied can
zero. This clearly does not happen, as demonstrated by be crucial. As the time over which they are compressed
the upper punch force values after maximum punch is reduced, such materials show less consolidation and
penetration in Fig. 8. This can only mean that as the this in turn can affect the physical strength of the tab-
upper punch ascends, the tablet expands elastically, let. For example, Armstrong and Palfrey[47] studied the
keeping in contact with the punch face. Thus, the area effects of punch speed change on the tablet strength of
ABC in Fig. 11 represents the work expended on com- four direct compression diluents. Though tablet
pressing the solid, and area DBC is the work delivered strength was reduced in all cases, solids such as modi-
back to the upper punch by the expanding tablet. The fied starch suffered the greatest reduction. Roberts and
difference between these two areas (i.e., ABD) has been Rowe investigated the relative sensitivity of the yield
termed the ‘‘Net work’’ of compression. pressures of a range of solids to changes in com-
Use of this technique has shown that the presence pression speed, and again found that those solids
of a granulating agent causes a marked increase in which deformed on compression showed the greatest
the plasticity of the powder mass, with a consequent change.[48]
increase in cohesion and tablet strength. The film of The rate of consolidation in an excentric press is
granulating agent between the particles can be governed by the speed at which the upper punch moves
regarded as a highly viscous liquid with a large yield into the die. This in turn is governed by the lengths of
value. Application of a force in excess of the yield value the excentric arms of the press and its speed of oper-
causes granules to deform, and this becomes perma- ation. In a rotary press, punch speed is governed by
nent when the force is reduced to below the yield the diameter of the die table, the diameter of the press-
value.[45] A somewhat similar mechanism is believed ure roll, the geometry of the punch head, and the speed
to account for the properties of some direct-com- of operation. Formulae for the calculation of punch
pression tablet diluents, e.g., spray-dried lactose, which speeds at any point of the compression cycle for both
consists of small crystalline masses embedded in an types of press have been derived.[49,50]
amorphous and more easily deformed matrix. It is important to distinguish the punch speed from
The force–displacement curve has been extensively the output of the press. Though rotary presses have a
used to characterize the compression event. However, much higher output than excentric presses, this largely
it depends on the availability of sensitive and properly derives from their multiplicity of punch stations. It
calibrated transducers. Even small inaccuracies in does not necessarily follow that their punch speeds
measuring displacement at high force values can have during compaction are significantly greater.[51]
a profound effect on the resultant value of the area If the work of compression in Joules is divided by
under the curve. Ragnarsson has provided a very the time in seconds over which compression occurs,
thorough review of the uses and potential pitfalls of then the power of compression (in Watts) is obtained.
the force–displacement curve.[46] An alternative method of calculating power of
Tablet Manufacture 3671

compression is to multiply the force by the punch 21. Shotton, E.; Leonard, G.S. Effect of intragranular and
speed when that force was applied. This too can be extragranular disintegrants on the particle size of disinte-
grated tablets. J. Pharm. Sci. 1976, 65 (8), 1170–1174.
obtained from the data in Fig. 8. This permits the com- 22. Lowenthal, W. Disintegration of tablets. J. Pharm. Sci.
pression events on different presses to be compared.[49] 1972, 61 (11), 1695–1711.
23. van Kamp, H.V.; Bolhuis, G.K.; de Boer, A.H.; Lerk, C.F.;
Lie-A-Huen, L. The role of water uptake on tablet disinte-
gration. Pharm. Acta Helv. 1986, 61 (1), 22–29.
ARTICLES OF FURTHER INTEREST 24. Malkowska, S.; Khan, K.A. Effect of recompression on the
properties of tablets prepared by dry granulation. Drug
Dev. Ind. Pharm. 1983, 9 (3), 331–347.
Drying and Dryers, p. 1435. 25. Bolhuis, G.K.; Chowhan, Z.T. Materials for direct com-
Tablet Manufacture by Direct Compression, p. 3673. pression. In Pharmaceutical Powder Compaction Tech-
nology; Alderborn, G., Nystrom, C., Eds.; Marcel Dekker,
Inc.: New York, 1996; 419–500.
26. Nystrom, C.; Karehill, P.G. Intermolecular bonding forces.
In Pharmaceutical Powder Compaction Technology; Alder-
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145–153. 42. Hersey, J.A.; Rees, J.E. Deformation of particles during
19. Salpekar, A.M.; Augsburger, L.L. Magnesium lauryl sulfate briqueting. Nature 1971, 230, 96.
in tabletting: effect on ejection force and compressibility. 43. Paronen, P.; Illka, J. Porosity-pressure functions. In Phar-
J. Pharm. Sci. 1974, 63 (2), 289–293. maceutical Powder Compaction Technology; Alderborn,
20. Shangraw, R.F.; Mitrevej, A.; Shah, M. A new era of tablet G., Nystrom, C., Eds.; Marcel Dekker, Inc.: New York,
disintegrants, pharmaceutical technology. 1980, Oct., 49–57. 1996; 55–75.
 3672 Tablet Manufacture

44. de Blaey, C.J.; Polderman, J. Compression of pharmaceuti- 48. Roberts, R.J.; Rowe, R.C. The effect of punch velocity on
cals. I the quantitative interpretation of force-displacement the compaction of a variety of materials. J. Pharm. Pharma-
curves. Pharm. Weekblad 1970, 105, 241–250. col. 1985, 37, 526–528.
45. de Blaey, C.J.; van Oudtshoorn, M.C.B.; Polderman, J. 49. Armstrong, N.A.; Abourida, N.M.A.H.; Gough, A.M. A
Compression of pharmaceuticals. III. Study on sulphadimi- proposed consolidation parameter for powders. J. Pharm.
dine. Pharm. Weekblad 1971, 106, 589–599. Pharmacol. 1983, 35, 320–321.
46. Ragnarsson, G. Force-displacement and network measure- 50. Rippie, E.G.; Danielson, D.W. Visco-elastic stress–strain
ments. In Pharmaceutical Powder Compaction Technology; behavior of pharmaceutical tablets: analysis during unload-
Alderborn, G., Nystrom, C., Eds.; Marcel Dekker, Inc.: ing and postcompression periods. J. Pharm. Sci. 1981, 70 (5),
New York, 1996; 77–97. 476–482.
47. Armstrong, N.A.; Palfrey, L.P. The effect of machine speed 51. Armstrong, N.A. Time-dependent factors involved in pow-
on the consolidation of four directly compressible tablet der compression and tablet manufacture. Int. J. Pharm.
diluents. J. Pharm. Pharmacol. 1989, 41, 149–151. 1989, 49, 1–13.
Tablet–Tablet
Tablet Manufacture by Direct Compression
Norman Anthony Armstrong
Welsh School of Pharmacy, Cardiff University, Cardiff, U.K.

INTRODUCTION As part of its complexity, wet granulation involves

the addition of a liquid (usually water), followed by
All tablets are made by compressing a particulate solid its removal, normally by evaporation. In addition to
between two punches in a die of a tablet press. For an the energy requirements of this drying process, the
active ingredient to be transformed into tablets of presence of water might bring about hydrolysis of

Tablet–Tablet
satisfactory quality, the formulation must have three the active ingredient, which will be exacerbated at the
essential attributes. elevated temperatures used for drying.
First, the formulation must flow into the die space of If a major component of the formulation such as the
the tablet press sufficiently rapidly and in a reproduc- diluent were to possess the necessary degrees of fluidity
ible manner. Otherwise, unacceptable variation in tab- and compressibility, granulation would be unnecessary.
let weight and content of active ingredient will ensue. This is the basis of the direct compression method of
Second, the particles in the formulation must cohere tablet manufacture.
when subject to a compressing force, and that coher- Prior to the early 1960s, there were very few
ence should remain after the compressive force has materials which possessed these properties. Little and
been removed. Mitchell[1] in their text Tablet Making, cite sodium
Third, after the compression event is complete, it chloride and bromide, potassium chlorate, bicarbonate
must be possible for the tablet to be removed from the and iodide, ammonium chloride, and hexamine as
press without damage to either the tablet or the press. having properties which permit tabletting without some
Very few active ingredients possess all three of these form of prior treatment. No reference is made by name
essentials and some possess none of them. Hence some to the process of direct compression.
preliminary treatment is almost invariably necessary. At about the same time, two materials were intro-
duced that were specifically designed to act as tablet
diluents and would not require preliminary treatment.
METHODS OF TABLET MANUFACTURES These were spray-dried lactose and microcrystalline
cellulose, introduced in 1962 and 1964, respectively.
There are three methods of tablet manufacture These two substances can be said to have initiated
designed to confer these essential attributes to a tablet the ‘‘direct compression revolution.’’ Since that time,
formulation. Wet granulation and direct compression a wide range of direct compression tablet diluents
are the most important, with dry granulation (also has become available. The properties of some of these
known as precompression or slugging) used in some materials will be reviewed later in this article.
circumstances. Fig. 1 shows the processes of wet granu- It is important to distinguish between true direct
lation and direct compression broken down into their compression diluents (i.e., excipients) and active ingre-
constituent stages. dients which are available in a direct compression
The relative simplicity of the direct compression form. These are usually high dose materials such as
process is immediately apparent. aspirin, paracetamol, and ascorbic acid. They can be
Ease of removal of the tablet from the press is, directly compressed into tablets, the only pretreatment
in theory at least, readily achieved. Friction occurs being mixing with a lubricant and perhaps a disintegrat-
between the tablet and the die and punches of the ing agent. However, such substances are more accurately
press, which can be overcome by including a lubricant described as ‘‘pregranulated’’ since the granulation
in the formulation. Hence every formulation, irrespec- process, either wet or dry, will have been carried out
tive of the method of manufacture, will include a by the excipient manufacturer. It is likely that such mate-
lubricant. This will usually be a metallic salt of a fatty rials will contain a binder. For example, ascorbic acid
acid such as magnesium stearate. pregranulated with either starch or hydroxypropyl cellu-
The other two prerequisites—flow and cohesion— lose is commercially available.[2]
can only be achieved by more elaborate techniques The perceived advantages of the direct compression
and are in fact the reasons why the wet and dry granu- process of tablet manufacture have given rise to a
lation processes were devised. considerable body of literature. Between 1970 and
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-120003785
Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved. 3673
 3674 Tablet Manufacture by Direct Compression

A B granulation and drying, and an ever-increasing use of

Active Active
automation have served to make wet granulation a much
ingredient ingredient more efficient and economic process than it once was.[3]
Diluent Mixing Diluent Nevertheless, the wet granulation process still retains
Lubricant Mixing many inherent disadvantages. Problems include choice
Binder Glidant and method of addition of the binder, and the effect of
Water Wetting Disintegrating drying time and temperature on drug stability and its
agent
distribution within the solid mass.
Compression
Granulation

Drying
Direct Compression Process: Advantages
and Disadvantages
Sieving
Lubricant
The most striking feature of the direct compression
Tablet–Tablet

Glidant Mixing
Disintegrating
process is its simplicity and hence economy. Less
agent Compression equipment is required and the number of stages in the
process, each of which will require validation, is greatly
Fig. 1 Comparison of the (A) wet granulation and (B) direct reduced. There are also lower labor costs, reduced
compression processes of tablet manufacture. processing time, and lower power consumption.
An important advantage of the direct compression
the end of 2000, there were 598 references to ‘‘direct process is that it is a dry procedure with no need for
compression’’ in the index of International Pharma- a drying stage. Thus, exposure to water and the ele-
ceutical Abstracts. It has been estimated that today, vated temperatures needed to remove that water are
some 40 years after the introduction of diluents specifi- avoided, resulting in a decreased risk of deterioration
cally designed for direct compression, about 50% of of the active ingredient.
worldwide tablet production is made by this method.[2] A further advantage of direct compression is that
The question must be asked why a process which has tablets disintegrate into their primary particles rather
so many apparent advantages and for which suitable than granular aggregates. The resultant increase in
materials seem to be plentifully available has not made surface area available for dissolution should result in
a greater impact. An innate conservatism in the phar- faster drug release.
maceutical industry is perhaps a factor, but cannot The primary limitation on the use of direct com-
be the complete answer. It is interesting to consider pression is that it depends on the fluidity and compress-
as an analogy that the process of film coating of ibility of a tablet diluent. Therefore, it cannot be used
tablets, coincidentally introduced at about the same for low potency, high dose active ingredients where
time as direct compression, has practically totally the inclusion of sufficient diluent in the formulation
replaced the sugar-coating technique. This shows that to permit direct compression would lead to unaccepta-
a new process can achieve significant and relatively bly large tablets. Thus, active ingredients such as para-
speedy penetration into an industrial environment if cetamol and aspirin do not lend themselves to the direct
it represents a major step forward. compression process. However, as stated earlier, such
ingredients are often available in pregranulated form.
Paradoxically, one of the root causes of difficulties
Wet Granulation Process: Advantages in the direct compression process is its simplicity. It
and Disadvantages must be regarded as a process in its own right, albeit
a simple one, rather than a simplified form of the wet
The wet granulation process is the traditional method granulation process. The key point to grasp is that
of manufacture and is frequently used in the pharma- wet granulation produces what is in effect a new raw
ceutical industry. Expertise in wet granulation is widely material, i.e., the granule. Minor variations in the
available, as is the required equipment. The process properties of the constituents of that granule are
improves flow and cohesion, reduces dust and cross- covered up by ‘‘submerging’’ them in a mass of binder.
contamination, and permits the handling of powder This is not so with direct compression. The properties
blends without loss of homogeneity. of each particle of every constituent remain essentially
Though it has been practiced for many years and unchanged. There is thus a greater need for within-
therefore may be perceived as an ‘‘old-fashioned’’ pro- batch and between-batch consistency.
cess, it must be borne in mind that the wet granulation There is also the possibility of segregation of the con-
process has itself undergone a transformation in recent stituents of the formulation after a homogeneous blend
decades. High-speed mixer–granulators, fluidized bed of active ingredient and excipients has been achieved.
Tablet Manufacture by Direct Compression 3675

In a wet granulated product, particles are stuck hydrogen bonds to give extremely strong tablets of
together by the binder and so there is a much reduced microcrystalline cellulose.
chance of segregation. Because segregation is princi- Most direct compression diluents are available from
pally a function of differences in particle size between only one source, but a few can be obtained from more
active ingredients and excipients, it is desirable that than one manufacturer. If multiple sources are avail-
the size of the direct compression diluent matches that able, they will be offered under individual registered
of the drug. This may not always be feasible. names. For example, microcrystalline cellulose is avail-
The simplicity of the direct compression process able under a number of brand names such as AvicelÕ
should obviously bring financial benefits. However, it (FMC Corporation), EmcocelÕ (Edward Mendell),
must be borne in mind that direct compression tablet and VivacelÕ (J. Rettenmaier). Chemical properties
diluents are considerably more expensive than conven- of such materials will be similar if not completely
tional diluents such as a-lactose monohydrate. identical, especially if there are pharmacopeial stan-
Regulatory considerations also play a part in a dards for identity and purity. However, it cannot be
decision whether or not to use the direct compression assumed that products from different manufacturers

Tablet–Tablet
process. Several years may elapse between the finalizing will have the similar physical properties which will
of a tablet formulation and its marketing. During this govern their performance in the tabletting process.
period, stability testing of the product will have occurred. Each brand will probably be accompanied by
The formulator must be confident that a chosen direct promotional literature to assist the sale of the product.
compression diluent will still be available for a consider- It is customary to describe its properties adjectivally,
able time after product marketing; otherwise reformula- e.g., ‘‘excellent flow,’’ ‘‘superior compressibility,’’ and
tion with all its attendant delay and expense will be to present data referring to that material alone. Hence
required. A number of direct compression diluents have comparison between different excipients can be some-
been marketed that were withdrawn after only a few times extremely difficult.
years because of lack of market penetration. Several authors have listed the attributes of the
Progression of product development will be accom- ‘‘ideal’’ direct compression diluent.[2,4] However, it
panied by scale-up of the manufacturing process of the must always be borne in mind that the diluent will
active ingredient to commercial proportions. This may invariably form part of a multicomponent mixture.
bring about changes in the physical properties of the At the very least, the diluent will be mixed with the
active ingredient. As stated earlier, the wet granulation active ingredient, and almost invariably a lubricant will
process can mask minor changes in physical properties, also be present. The greater the proportion of active
but this masking cannot occur in direct compression. ingredient in the formulation, the less influence the
For these reasons, direct compression has been most diluent will have on the properties of the tablet.
widely adopted by manufacturers of generic (i.e., non- One of the difficulties that beset the product devel-
innovative) pharmaceuticals. During the time when the oper is the lack of meaningful tests by which excipients
active ingredient is covered by patent protection, (including direct compression diluents) can be assessed.
its optimum manufacturing process will have been This has led to the development of the so-called ‘‘func-
achieved, and so subsequent batch-to-batch variation tionality tests.’’ Some functionality tests that have been
in its physical properties ought then to be minimal. suggested (e.g., particle size, surface area) are in fact
physical test methods being carried out under closely
defined conditions.[5] The relation of such a test to the
FACTORS INFLUENCING THE CHOICE OF A actual function of the excipient needs to be established.
DIRECT COMPRESSION TABLET DILUENT After this link has been made, a more suitable title for
these tests might be ‘‘functionality-related tests.’’[6]
A wide range of substances is or has been marketed as Nevertheless, it is useful to consider some of the
direct compression tablet diluents. In general, these are desirable properties of a direct compression diluent and
commonly occurring materials whose properties have how these properties can be appropriately measured.
been modified in such a way to give the fluidity and
compressibility demanded by the direct compression
process. Many direct compression diluents are aggre- Properties Required of a Direct
gates of primary particles. For example, an aqueous Compression Diluent
slurry of a-lactose monohydrate is spray-dried to
give an agglomerated product that flows better and is Fluidity
more compressible than the parent substance. A
second example is the acid hydrolysis of a-wood Good flow is a prerequisite for any tablet formulation to
cellulose to yield particles containing bundles of micro- ensure uniformity of tablet weight, which in turn contri-
crystals of cellulose. These can cohere by means of butes to uniformity of content. Flow can be measured by
 3676 Tablet Manufacture by Direct Compression

methods such as angle of repose, flow through an orifice, Table 1 Some units used for the construction of tablet
and by using flow cells, but more meaningful data can be strength profiles
obtained by measuring the uniformity of weight of the Abscissa Ordinate
tablets themselves. Flow properties can often be
improved by the inclusion of a glidant in the formulation. Parameter Unit Parameter Unit
Force kg Crushing strength kg
lb (hardness) Strong-Cobb
Ease of mixing and lack of segregation
kN units
N N
Achievement of a homogeneous mixture of active kp
ingredient and diluent is essential to obtain tablets with
Pressure kg cm
2 Tensile strength kg cm
2
an acceptable uniformity of content of active ingredi- Pa Pa
ent. As stated earlier, there is a risk of segregation in Mpa MPa
a direct compression mixture, because the components lb in.
2
Tablet–Tablet

are not stuck together as they are in wet granulation. (From Ref.[9].)
The main cause of segregation is differences in the
particle size of components, with differences in shape
and density being secondary factors.[7] Hence it is where Ts is the tensile strength of the tablet (MPa), P is
desirable that there should not be differences between the crushing strength (N), d is the tablet diameter (m),
the particle sizes of active ingredient and diluent. It is and t is the tablet thickness (m).
unlikely that the size of the active ingredient particles Use of this equation presupposes that the tablet is
can be changed to match those of the diluent, so the circular in cross-section and of uniform thickness,
reverse is desirable. Thus, the ideal direct compression i.e., it is cylindrical. Pitt et al. have attempted to extend
diluent should be available in a range of sizes. the concept of tensile strength to tablets which are not
An alternative approach is to use the concept of cylindrical.[11]
ordered mixing in which fine particles of the active Even though comparison is often difficult, it would
ingredient are dispersed over the surface of much lar- be rendered considerably easier if the units used
ger diluent particles.[8] This is only feasible with potent conformed to the SI system of weights and measures.
drugs when the diluent will comprise by far the major Thus, the meter, newton, and pascal should be used
component of the formulation. as the units of length, force, and pressure, respectively.

Compression pressure–tablet strength profile Capacity or dilution potential

This is the relationship between the compression By definition, direct compression diluents are intended
pressure applied to the formulation and the physical to be mixed with other ingredients. Therefore, not
strength of the resulting tablets. It would seem to be a only should the pressure–tablet strength profile of the
most important piece of information, yet its derivation diluent be determined, but also should those of
is by no means straightforward. Also comparison of mixtures of the diluent with an active ingredient.
data derived by different researchers is often difficult. The capacity of a direct compression diluent is the
If such data are presented graphically, either force proportion of another ingredient that can be mixed
or pressure can be used as the abscissa. Unless the with it while still obtaining tablets of acceptable qual-
cross-sectional dimensions of the tablet are known, ity. The definition of ‘‘acceptable’’ will depend on the
interconversion between force and pressure is impos- purpose for which the tablets are required.
sible. Furthermore, a wide variety of units of force The magnitude of the effect that a given active
and pressure have been used (Table 1), which again ingredient will have on tablet properties will clearly
makes comparisons difficult if not impossible. depend on the tabletting properties of that substance.
The physical strength of the tablet usually forms the If it is also capable of direct compression, then the effect
ordinate of a graph. The variety of units used for this is will not be great. If, however, it is a substance that is dif-
also shown in Table 1. Here too interconversion can be ficult to compress into tablets, then it will cause a
extremely difficult. The breaking strength of a tablet is marked deterioration in tablet quality when mixed with
dependent on its physical dimensions. It is therefore the diluent. Therefore, for a reliable test of capacity, the
logical to use some measure of strength that is inde- direct compression diluent should be mixed with a
pendent of tablet size. Therefore the tensile strength ‘‘standard’’ substance and tabletted under standard-
of the tablet is often calculated according to Eq. (1).[10] ized conditions. The pressure–strength profiles of the
mixtures can then be constructed. Paracetamol[12,13]
T s ¼ 2P=Pdt ð1Þ and ascorbic acid[9] have been used as standards.
Tablet Manufacture by Direct Compression 3677

Reworking

A faulty batch of tablets can sometimes be recovered

Tensile by grinding up the tablets and recompressing them, a
A process which is known as reworking and is analogous
strength
(MPa) to the dry granulation method of tablet manufacture.
This can sometimes cause problems with a direct com-
pression formulation. Many direct compression diluent
B particles are in the form of aggregates, e.g., spray-dried
lactose is composed of small crystals of lactose embed-
0 100 200 300 400 ded in amorphous lactose. If these aggregates are
Compression pressure (MPa) compressed, their structure may be broken down to
such an extent that subsequent recompression will
Fig. 2 Calculation of the capacity of a direct compression result in impaired tablet quality.

Tablet–Tablet
diluent. (From Ref.[9].)
The technique of Malkowska and Khan,[14] used as
described before to determine the capacity of a direct
A method of quantitatively assessing the capacity of a compression diluent, was originally developed as a
direct compression diluent has been devised by Minchom method of expressing the ability of a formulation to be
and Armstrong[12] who adapted a concept originally reworked. Referring to Fig. 2, the upper curve represents
introduced by Malkowska and Khan.[14] Tablets contain- the strength of tablets prepared without reworking and
ing only diluent were prepared over a range of com- the lower curve is the strength of reworked tablets. The
pression pressures, their strengths were measured, and a reworking index is calculated from the ratio of the areas
pressure–strength profile was constructed. A mixture of under the curves as described previously.
diluent plus active ingredient was then compressed under
identical conditions, tablet strengths were determined, The mechanism of consolidation
and the profile was constructed as before (Fig. 2).
The profiles were then fitted by regression analysis A fundamental property of a solid is the mechanism by
to quadratic equations, and integration of these equa- which it consolidates under the influence of a com-
tions between limits (in this case pressures of 100 and pressing force. There are two principal mechanisms—
300 MPa) gave the area under the curves. Thus, the fragmentation and deformation—though most solids
change in tablet strength brought about by the will show a mixture of the two with one mechanism
addition of the active ingredient is given by the ratio predominating. The mechanism can have a major
B/(A þ B). Details of the application of this method influence on tablet properties.
to mixtures of direct compression lactose and ascorbic The effect of compression speed on tablet quality is
acid are shown in Table 2.[9] The technique has been dependent on the consolidation mechanism. Fragmen-
developed further by Habib et al.[15] tation can be regarded as a virtually instantaneous pro-
The effect of the active ingredient on flow properties cess. Thus, solids which consolidate by fragmentation
can be determined at the same time by calculating the show little dependence, if any, on the speed at which
uniformity of weight of the tablets used to prepare the consolidation pressure is applied. Deformation on
pressure–strength profiles. the other hand is time-dependent. It takes a finite time
for deformation to occur, and at high rates of punch
movement, not enough time may be available for the
Table 2 Areas under pressure–tablet
full effect of the pressure to be exerted. Changes in
strength profiles for mixtures of ascorbic
punch speed can arise by changing the speed of the tab-
acid and direct compression lactitol
let press or by changing from a relatively slow-speed
Ascorbic acid excentric press to a high-speed rotary. It must be
concentration (%) Area ratio
stressed that the key parameter is punch speed rather
0 1.00 than production rate.[16] Roberts and Rowe[17] derived
20.0 0.75 a parameter that they called the strain rate sensitivity.
33.3 0.47 This classifies substances according to how tablet strength
42.9 0.35 is affected by changes in punch speed. Details of some of
the substances investigated by Roberts and Rowe are
50.0 0.32
shown in Table 3, and many of these are direct com-
55.8 0.27 pression diluents.
60.0 0.20 The practical implications of this work are shown
(From Ref.[9].) in Table 4.[18] As punch speed is increased, the strength
 3678 Tablet Manufacture by Direct Compression

Table 3 Strain rate sensitivities of powders incorporation should be stated in any publication
Strain rate relating to direct compression diluents.
Substance sensitivity (%)
Calcium phosphate 0
TABLET DILUENTS USED IN
Calcium carbonate 0
DIRECT COMPRESSION
Heavy magnesium carbonate 0
Paracetamol DC 1.8 A wide variety of materials have been used as direct
Paracetamol 10.6 compression diluents, and this has given rise to a con-
Lactose 16.2 siderable literature. Many research reports originate
Tablettose 19.2 with organizations marketing a specific diluent, or
are derived from work sponsored by that organization.
Anhydrous lactose 20.3
Such publications will obviously report data and results
Avicel PH101 38.9 relevant to that one diluent.
Tablet–Tablet

Sodium chloride 39.9 However, there are some publications which seek to
Mannitol 46.4 compare a range of diluents, and these are particularly
Maize starch 49.3 valuable.[20,21] Perhaps, the most comprehensive review
(From Ref. [17]
.) of direct compression diluents has been published by
Bolhuis and Chowhan.[2] Monographs of many direct
compression diluents are to be found in the Handbook
of Pharmaceutical Excipients, which contains details
of tablets of dicalcium phosphate is only marginally
of the physical properties of these substances.[22]
affected, whereas those made of starch suffer a severe
Direct compression diluents are often commonly
reduction in their strength. As punch speed is increased,
occurring substances which have been physically
speed-sensitive materials give a progressively more
modified to give the necessary degree of fluidity and
porous and hence weaker tablet. It follows that in any
compressibility. They are most conveniently classified
report on the tabletting properties of a diluent, the type
according to their source, viz.:
of press and its speed of operation should be stated.
A second area in which consolidation mechanism is
 Cellulose and cellulose derivatives.
important is in the sensitivity of diluents to the effects
 Inorganic materials.
of lubricants. In general, addition of a lubricant such
 Polyols.
as magnesium stearate causes a reduction in tablet
 Starch and starch derivatives.
breaking strength. As the diluent is mixed with the
 Sugars.
lubricant, each diluent particle becomes coated with a
 Mixtures and coprocessed products.
thin film of lubricant which interferes with interparti-
culate bonding. However, if fragmentation is the
primary method of consolidation, new surface that is
Information on some direct compression diluents is
uncontaminated by lubricant is continually generated,
given in Table 5.
and so bonding is less affected. Thus, factors which
affect the distribution of the lubricant over the diluent
surface may have an influence on tablet strength, the Cellulose and Cellulose Derivatives
magnitude of which depends on the predominant
consolidation mechanism. Such factors include mixer Since its introduction as a direct compression diluent
design and mixing time and speed.[19] It follows in 1964, microcrystalline cellulose has become a phar-
that the lubricant, its concentration, and method of maceutical excipient of great importance. It consists

Table 4 Tablet tensile strengths (MPa) produced by a compression pressure of 80 MPa on an

excentric press
Press speed Dicalcium Microcrystalline
(tablets per minute) phosphate Lactose cellulose Starch 1500
20 0.63 0.77 5.4 0.28
40 0.62 0.76 5.2 0.25
160 0.60 0.68 3.9 0.15
% reduction 5.0 12 28 47
(From Ref.[18].)
Tablet Manufacture by Direct Compression 3679

Table 5 Direct compression diluents, their proprietary names, and their manufacturers
Diluent Proprietary name Manufacturer
Cellulose and cellulose derivatives
Cellulose, microcrystalline AvicelÕ FMC
EmcocelÕ Penwest
VivacelÕ Rettenmaier
Cellulose, powdered Solka-FlocÕ Degussa
ElcemaÕ Penwest
Cellulose, silicified microcrystalline ProsolvÕ Penwest
Inorganic materials
Calcium phosphate, dibasic anhydrous Dicafos-AÕ Budenheim
Calcium phosphate, dibasic dihydrate CalstarÕ FMC
DicafosÕ Budenheim

Tablet–Tablet
EmcompressÕ Penwest
Di-TabÕ Stauffer
Calcium phosphate, tribasic TricafosÕ Budenheim
TricompressÕ Penwest
Tri-TabÕ Rhone-Poulenc
Calcium sulphate CompactrolÕ Penwest
Polyols
Xylitol XylitabÕ Xyrofin
Õ
Lactitol Finlac Xyrofin
Mannitol PearlitolÕ Roquette
Õ
Sorbitol Neosorb Roquette
Sorbitol InstantÕ Merck
Starch and starch derivatives
Starch, pregelatinized Starch 1500Õ
Starx 1500Õ Colorcon
Modified rice starch PrimotabÕ Avebe
Granulated corn starch Sepistab ST200 Seppic
Sugars
Compressible sugar Destab Desmo
Dipac Amstar
Nutab Ingredient
Technology
Sugartab Penwest
Dextrates EmdexÕ Penwest
CelutabÕ Penick and Ford
Lactose
Agglomerated Tablettose Meggle
Pharmatose DCL15Õ DMV
Anhydrous a Pharmatose DCL30Õ DMV
Õ
Anhydrous b Pharmatose DCL21 DMV
Anhydrous DT Sheffield
Spray-dried Fast-FloÕ Foremost
ZeparoxÕ Borculo
Pharmatose DCL11Õ DMV
Mixtures and coprocessed products
Anhydrous lactose–anhydrous lactitol Pharmatose DCL40Õ DMV
Calcium sulphate–microcrystalline cellulose Cel-O-Cal FMC
Lactose–cellulose CellactoseÕ Meggle
Lactose–starch StarlacÕ Roquette
Lactose–povidone LudipressÕ BASF
 3680 Tablet Manufacture by Direct Compression

of aggregates of microcrystals and is isolated from principally by fragmentation, so although a lubricant

a-wood cellulose by acid hydrolysis. is needed, tablet strength loss is low.[19] It is non-
When compressed, microcrystalline cellulose particles hygroscopic at humidities up to 80%, and due to its
deform plastically and the surfaces thus brought into hydrophilic nature, dicalcium phosphate dihydrate
contact unite by hydrogen bonding. Because tablets tablets are rapidly penetrated by water on immersion.
made from microcrystalline cellulose are extremely Despite this, the tablets do not disintegrate because they
strong, there is a high dilution potential and they can are almost insoluble in water.[27] Dicalcium phosphate
withstand weakening caused by lubricants.[19] In fact, dihydrate is, however, soluble in acidic media such as
microcrystalline cellulose tablets exhibit such a low gastric juice.
coefficient of friction that they may need no lubricant. The water of hydration is relatively easily lost from
Fluidity of microcrystalline cellulose is low and the dibasic calcium phosphate dihydrate, and this may
addition of a glidant may be necessary. Its bulk density have consequences for the stability of products contain-
is also low. Unlike many direct compression diluents, it ing it.[28]
is available in a wide range of particle sizes. Anhydrous dibasic calcium phosphate and calcium
Tablet–Tablet

Microcrystalline cellulose is available from several triphosphate can also be used for direct compression.
different sources. These can exhibit a range of table- The latter is actually a mixture of calcium phosphates
tting properties, and so the substitution of one brand including tricalcium orthophosphate [Ca3(PO4)2] and
of microcrystalline cellulose by another must be hydroxyapatite [Ca5(OH)(PO4)3]. The preparation and
approached with caution.[23] Microcrystalline cellulose properties of calcium phosphates have been reviewed
is quite hygroscopic, and its tabletting properties are by Carstensen and Ertell,[29] and their tabletting proper-
dependent on its moisture content, with water causing ties have been studied by Bryan and McAllister.[30]
the interparticulate hydrogen bonds to weaken. There- A direct compression diluent based on calcium
fore, comparisons between different brands must also sulphate is also available.[31]
take the moisture content into account.[24]
Powdered cellulose has been used as a direct com-
pression diluent. Though it forms hard tablets, fluidity Polyols
is poor and dilution potential is low. Like microcrystal-
line cellulose it has some self-lubricating properties, Several solid polyols can be used as direct compression
but addition of a lubricant is usually necessary, causing diluents, usually after some physical modification. Most
a marked reduction in tablet strength.[25] such polyols can be obtained from natural sources, but
Silicified microcrystalline cellulose is a coprocessed are usually manufactured by hydrogenation of the
mixture of microcrystalline cellulose and 2% colloidal parent sugar molecule. Some properties of polyols,
silicon dioxide, which has improved flow and binding together with comparative data for lactose and sucrose,
properties compared to microcrystalline cellulose itself.[26] are shown in Table 6.

Sorbitol
Inorganic Materials
This is probably the most commonly used polyol in
The most widely used inorganic direct compression tablet manufacture. It is freely soluble in water,
diluent is calcium phosphate. It is available in several producing strong tablets. Its negative heat of solution,
forms, but the unmilled (i.e., coarse) dibasic dihydrate which produces a cooling effect in the mouth, makes it
(CaHPO4  2H2O) is the most frequently used. It has particularly useful in chewable tablets and ‘‘sugar-
good flow and binding properties. Consolidation is free’’ confectionary. Furthermore, most polyols are

Table 6 Properties of some polyols

Water content at
Solubility in water Melting point Heat of solution Sweetness
(g per 100 cm3) ( C) (J g
1) 55% RH 75% RH 100% RH (sucrose = 100)
Lactitol 170 180
54 0.0 0.1 37.2 35
Mannitol 18 167
121 0.5 0.5 83.7 50
Sorbitol 200 110
113 6.4 10.4 122.2 60
Xylitol 160 94
153 0.1 0.1 70.6 100
Lactose 22 201
18 0.8 0.8 7.0 15
Sucrose 200 160
17 1.0 1.0 38.0 100
Tablet Manufacture by Direct Compression 3681

non-cariogenic, which increases their usefulness in this amylose, and 15% free amylopectin. Though it has
area. It is hygroscopic, and shows an appreciable been described as a direct compression diluent,[38]
increase in moisture content with attendant impairment tablets made from pregelatinized starch show low
of flow properties when exposed to relative humidities physical strength. The principal application of pregela-
in excess of 65%. At very high humidities, sorbitol tinized starch in tablet formulation is as a disintegrat-
absorbs enough water to bring about dissolution. ing agent. It retains the disintegrating ability of natural
Sorbitol can exist in four crystalline forms. Guyot- starch without the deleterious effects on flow and
Hermann, Leblanc, and Draguet-Brugmans[32] com- tablet strength that natural starch would bring about.
pared 11 commercially available varieties of sorbitol,
and found three of these four forms to be present.
g-Sorbitol was found to be the most useful as a tablet Sugars
diluent. The method of manufacture has also been
shown to affect tabletting properties, differences being Lactose
attributed to variations in particle shape and surface

Tablet–Tablet
properties. Spray-dried varieties of sorbitol are avail- Lactose is a naturally occurring disaccharide contain-
able as direct compression diluents which are claimed ing one galactose unit and one dextrose unit. It is a
to have overcome problems associated with the differ- constituent of all forms of mammalian milk, but is
ent crystalline forms.[33] produced commercially from cow’s milk, usually as a
by-product of the cheese industry. Lactose can exist
Mannitol in two isomeric forms, a-lactose and b-lactose, and
can be either crystalline or amorphous. Crystalline
Mannitol is an isomer of sorbitol. Like the latter, it has a-lactose occurs in both monohydrate and anhydrous
a negative heat of solution which makes it a useful forms, but b-lactose only exists in the anhydrous form.
excipient for chewable tablets and lozenges. It is less The temperature of crystallization is the principal
hygroscopic than sorbitol and has about one-tenth of determinant of which form is obtained.[39]
the solubility in water. Similarly to sorbitol, several Though crystalline a-lactose monohydrate is the most
polymorphic forms are available which differ in their common tablet diluent, it is usually used in granulated
ability to form tablets.[34] However, unmodified manni- rather than in direct compression formulations. Neither
tol cannot be used for direct compression because of its flow properties nor its binding properties are good
poor flow and binding properties. Directly compress- enough to form satisfactory tablets without preliminary
ible forms are available in a range of particle sizes treatment. Bonding properties are improved by conver-
which are reported to produce excellent tablets. sion into aggregates of a-lactose monohydrate crystals
by fluid bed granulation. This product is virtually free
Lactitol and xylitol from amorphous lactose.[40]
Spray-dried lactose was the first direct compression
These are both commercially available in forms suit- diluent to be introduced.[41] It had a major impact on
able for direct compression with good flow and binding tabletting technology and it is still widely used. Spray
properties. The former is a water-granulated product drying an aqueous suspension of lactose yields a
of microcrystalline aggregates.[35] A similar form of product that largely consists of crystals of a-lactose
xylitol is available, and in the case of xylitol, there monohydrate (about 80%) held together in a glass of
are also products pregranulated with either polydex- amorphous material (about 20%). Spray-dried lactose
trose or carboxymethyl cellulose.[36] Both substances exhibits excellent flow properties due to the spherical
are highly soluble in water with negative heats of shape of the aggregates. However, its ability to form
solution. strong tablets is limited and it has low dilution poten-
tial, so it is primarily used in tablets in which it forms
the major ingredient. Fragmentation is the major
Starch and Starch Derivatives consolidation mechanism, and so tablet strength is
not significantly affected by lubricants. Spray-dried
Starch is a very widely used tablet excipient, but in its lactose can be obtained from several manufacturers
natural state, it does not possess the fluidity and bind- whose products differ slightly. A comparative study
ing characteristics needed as a tablet diluent. The of spray-dried lactoses from a number of sources has
major consolidation mechanism of starch is by defor- been published by Whiteman and Yarwood.[21]
mation with a high elastic component.[37] In addition, Anhydrous lactose is primarily anhydrous b-lactose
starch shows a high degree of lubricant sensitivity.[19] with up to about 25% anhydrous a-lactose. It consists of
Pregelatinized starch, often referred to as Starch agglomerates of fine crystals produced by roller drying a
1500, contains about 80% unmodified starch, 5% free solution of a-lactose monohydrate. Flow properties are
 3682 Tablet Manufacture by Direct Compression

good, and tablet strength was found to be superior to whereas the fluidity of microcrystalline cellulose is
other lactose products.[21] Anhydrous b-lactose is poor yet it forms extremely strong tablets. It is under-
much more soluble than the a-isomer, and extended standable, therefore, that the possibilities of combining
disintegration times of tablets made from anhydrous diluents have been considered, the aim being to com-
lactose have been attributed to the presence of bine the advantages of both components without their
anhydrous a-lactose in the roller dried product.[42] disadvantages.
Anhydrous a-lactose can be produced by thermal The compaction properties of mixtures have been
or chemical dehydration of a-lactose monohydrate. reviewed by Fell,[45] who concluded that the relation-
During this process, the starting material changes from ship between the tabletting properties of a mixture
single crystals to aggregates of anhydrous a-lactose could only rarely be predicted from knowledge of
particles. Flowability and binding properties are good, the same properties of the individual components.
but slow dissolution of tablets made from anhydrous Nevertheless, some success has been achieved.[46]
a-lactose has proved a major limitation to its use.[42] In recent years, a number of coprocessed excipient
combinations have been marketed, which undoubtedly
Tablet–Tablet

Sucrose possess advantages over physical mixtures of the same

components. Such materials include mixtures of
A non-reducing disaccharide obtained from vegetable lactose and povidone,[47] cellulose and lactose,[48] and
sources, sucrose is a widely used pharmaceutical excipi- anhydrous lactose and anhydrous lactitol. A disadvan-
ent. Because it is more hygroscopic than lactose, it is tage of this approach is that the relative proportions of
used less in solid dosage forms. The compactability the components are fixed, and such a combination may
of pure sucrose is poor, but modified forms of sucrose not be universally optimal.
for direct compression are available. These are collec-
tively termed ‘‘compressible sugar,’’ and may contain,
depending on source, starch, maltodextrin, or invert
sugar, together with a lubricant. Several different types REFERENCES
of compressible sugar have been compared.[43] The
1. Little, A.; Mitchell, K.A. Tablet Making, 2nd Ed.; Northern
minor components obviously played a major role in Publishing: Liverpool, U.K., 1963.
tablet formation, since significant differences were 2. Bolhuis, G.K.; Chowhan, Z.T. Materials for direct
obtained between varieties. Compressible sugar is often compression. In Pharmaceutical Powder Compaction
Technology; Alderborn, G., Nystrom, C., Eds.; Marcel
used for lozenges and chewable tablets—because of Dekker, Inc.: New York, 1996; 419–500.
the high solubility of sucrose, tablets tend to dissolve 3. Kristensen, H.G.; Schaefer, H.G.; Granulation, T. Drug
rather than disintegrate. Dev. Ind. Pharm. 1987, 13 (4/5), 803–872.
4. Sheth, B.B.; Bandelin, F.J.; Shangraw, R.F. Compressed
tablets. In Pharmaceutical Dosage Forms: Tablets, 1st Ed.;
Dextrose and dextrates Lieberman, H.A., Lachman, L., Eds.; Marcel Dekker, Inc.:
New York, 1980; Vol. 1, 109–185.
5. Brittain, H.G. Functionality testing of excipient materials.
Dextrose does not lend itself to direct compression. Pharm. Technol. 1993, 9 (9), 72–73.
However, a spray-crystallized product, the major 6. Armstrong, N.A. Functionality related tests for excipients.
constituent of which is dextrose, is used in direct com- Int. J. Pharm. 1997, 155, 1–5.
7. Campbell, H.; Bauer, W.C. Cause and cure of demixing in
pression. This is known as dextrates, and is produced solid–solid mixers. Chem. Eng. 1966, 179–185.
by the partial hydrolysis of starch. It consists of about 8. Staniforth, J.N.; Rees, J.E.; Kayes, J.B. Relation between
90% dextrose, together with about 5% maltose and mixing time and segregation of ordered mixes. J. Pharm.
Pharmacol. 1981, 33, 175–176.
higher polysaccharides.[44] Though both hydrated and 9. Armstrong, N.A. Selection of excipients for direct com-
anhydrous forms of dextrates have been described, pression tablet formulations. Pharm. Technol. Eur. 1997,
only the former is commercially available. Dextrose 24–30.
10. Fell, J.T.; Newton, J.M. Determination of tablet strength by
is freely soluble in water and is highly hygroscopic, the diametral compression test. J. Pharm. Sci. 1970, 59 (5),
and hence its use in atmospheres of high humidity 688–691.
may cause problems. Because of its sweet taste and 11. Pitt, K.G.; Newton, J.M.; Richardson, R.; Stanley, P.
Material tensile strength of convex-faced aspirin tablets. J.
negative heat of solution, it is recommended for use Pharm. Pharmacol. 1989, 41, 289–292.
in chewable tablets. 12. Minchom, C.M.; Armstrong, N.A. A proposed technique
for expressing the capacity of directly compressible tablet
diluents. J. Pharm. Pharmacol. 1989, 39, 69.
13. Wells, J.I.; Langridge, J.R. Dicalcium phosphate dihydrate–
Mixtures and Coprocessed Products microcrystalline cellulose systems in direct compression
tabletting. Int. J. Pharm. Tech. and Prod. Manuf. 1981,
Though numerous direct compression diluents are 2 (2), 1–8.
14. Malkowska, S.; Khan, K.A. The effect of recompression on
available, none is ideal. For example, spray-dried the properties of tablets prepared by dry granulation. Drug
lactose flows easily but forms relatively weak tablets, Dev. Ind. Pharm. 1983, 9 (3), 331–347.
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15. Habib, Y.; Augsburger, L.; Reier, G.; Wheatley, T.; phenobarbitone sodium tablets prepared with compactrol.
Shangraw, R. Dilution potential: a new perspective. Pharm. Drug Dev. Ind. Pharm. 1985, 11 (11), 1947–1955.
Dev. Technol. 1996, 1 (2), 205–212. 32. Guyot-Hermann, A.M.; Leblanc, D.; Draguet-Brugmans,
16. Armstrong, N.A. Time-dependent factors involved in pow- M. Gamma sorbitol as a diluent in tablets. Drug Dev.
der compression and tablet manufacture. Int. J. Pharm. Ind. Pharm. 1985, 11 (2/3), 551–564.
1989, 49, 1–13. 33. Basedow, A.M.; Moschl, G.A.; Schmidt, P.C. Sorbitol
17. Roberts, R.J.; Rowe, R.C. The effect of punch velocity on instant, an excipient with unique tabletting properties. Drug
the compaction of a variety of materials. J. Pharm. Pharma- Dev. Ind. Pharm. 1986, 12 (11–13), 2061–2089.
col. 1985, 37, 526–528. 34. Debord, B.; Lefebre, C.; Guyot-Hermann, A.M.; Hubert, J.;
18. Armstrong, N.A.; Palfrey, L.P. The effect of machine speed Bouche, R.; Guyot, J.C. Study of different crystalline forms
on the consolidation of four directly compressible tablet of mannitol: comparative behaviour under compression.
diluents. J. Pharm. Pharmacol. 1989, 41, 149–151. Drug Dev. Ind. Pharm. 1987, 13 (9–11), 1533–1546.
19. Bolhuis, G.K.; Holzer, A.W. Lubricant sensitivity. In Phar- 35. Armstrong, N.A. Direct compression characteristics of lac-
maceutical Powder Compaction Technology; Alderborn, titol. Pharm. Tech. Eur. 1998, 10 (2), 42–46.
G., Nystrom, C., Eds.; Marcel Dekker, Inc.: New York, 36. Morris, L.E.; Moore, J.C.; Schwartz, J.B. Characterisation
1996; 517–560. and performance of a new direct compression excipient
20. Bolhuis, G.K.; Reichman, G.; Lerk, C.F.; van Kamp, H.V.; for chewable tablets: xylitab. Drug Dev. Ind. Pharm.
Zuurman, K. Evaluation of anhydrous a-lactose, a new 1996, 22 (9/10), 925–932.

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excipient in direct compression. Drug Dev. Ind. Pharm. 37. Paronen, P.; Juslin, M. Compression characteristics of four
1985, 11 (8), 1657–1681. starches. J. Pharm. Pharmacol. 1983, 35, 627–635.
21. Whiteman, M.; Yarwood, R.J. The evaluation of six 38. Manudhane, K.S.; Contractor, A.M.; Kim, H.Y.; Shangraw,
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22. Kibbe, A., Ed.; Handbook of Pharmaceutical Excipients, 39. Lerk, C.F.; Andreae, A.C.; de Boer, A.H.; de Hoog, P.;
3rd Ed.; Pharmaceutical Press: London, 2000. Kussendrager, K.D.; van Leverink, J. Transition of lactoses
23. Doelker, E.; Mordier, D.; Iten, H.; Humbert-Droz, P. by mechanical and thermal treatment. J. Pharm. Sci. 1984,
Comparative tabletting properties of sixteen microcrystal- 73, 857–859.
line celluloses. Drug Dev. Ind. Pharm. 1987, 13 (9–11), 40. Shangraw, R.F.; Wallace, J.W.; Bowers, F.M. Morphology
1847–1875. and functionality of tablet excipients for direct compression.
24. Roberts, R.J.; Rowe, R.C. Source and batchwise variability Pharm. Tech. 1981, 5, 69–78.
in the compressibility of microcrystalline cellulose. J. 41. Gunsel, W.C.; Lachman, L. Comparative evaluation of
Pharm. Pharmacol. 1987, 39, 70P. tablet formulations prepared from conventionally processed
25. Bos, C.E.; Vromans, H.; Lerk, C.F. Lubricant sensitivity in and spray-dried lactose. J. Pharm. Sci. 1963, 52 (2), 178–182.
relation to bulk density for granulations based on starch 42. Van Kamp, H.V.; Bolhuis, G.K.; Kussendrager, K.D.;
and cellulose. Int. J. Pharm. 1991, 67, 39–49. Lerk, C.F. Studies on tabletting properties of lactose. IV.
26. Sherwood, B.E.; Hunter, E.A.; Staniforth, J.N. Silicified Dissolution and disintegration properties of different types
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stabilisation of the tablet excipient calcium hydrogen Nystrom, C., Eds.; Marcel Dekker, Inc.: New York, 1996;
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2031–2055. 46. Armstrong, N.A.; Lowndes, D.H.L. The use of mixtures of
29. Carstensen, J.T.; Ertell, C. Physical and chemical properties spray-dried lactose and microcrystalline cellulose as direct
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30. Bryan, J.W.; McCallister, J.D. Matrix forming capabilities 47. Schmidt, P.C.; Rubensdorfer, C.J. Evaluation of ludipress
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 Tablet Press Instrumentation
Michael Levin
Metropolitan Computing Corporation, East Hanover, New Jersey, U.S.A.

INTRODUCTION A new generation of ‘‘compaction simulators’’ was

born when a mechanical press replicator was patented
This article is designed to facilitate the understanding in the year 2000.[23]
of the general principles of tablet press instrumentation A good exposé of the early stages of press instru-
Tablet–Tablet

and the benefits thereof by the formulators, process mentation and resulting research is presented in review
engineers, validation specialists, and quality assurance articles by Schwartz,[24] Marshall,[25] and Jones.[26] A
personnel, as well as production floor supervisors comprehensive review can be found in a relatively
who would like to understand the basic standards recent paper by Çelik and Ruegger.[27]
and techniques of getting information about their For other historical information on the press
tableting process. instrumentation, the reader is encouraged to peruse
Ridgeway Watt’s[28] volume. There have been a
number of papers published on various disparate
instrumentation topics, and in a recent volume by
Muños-Ruiz and Vromans,[29] there are two good arti-
HISTORY OF TABLET PRESS cles on the subject—but, unfortunately, they deal with
INSTRUMENTATION marginal issues of single station press and instrumen-
ted punch only.
In 1952–1954 Higuchi and his group[1] have instrumen-
ted upper and lower compression, ejection, and punch
displacement on an eccentric tablet press, and pio-
neered the modern study of compaction process. This
DATA ACQUISITION PRINCIPLES: FROM
work was followed by Nelson[2,3] who was the member
TRANSDUCERS TO COMPUTERS
of the original group.
In 1966, a U.S. patent was granted to Knoechel
To monitor and control a tablet press, certain sensors
and co-workers[4] for force measurement on a tablet
must be installed at specific locations on the machine.
press. This patent was followed by two seminal articles
These sensors are called transducers. In general, a
in Journal of Pharmaceutical Sciences on the practical
transducer is a device that converts energy from one
applications of instrumented rotary tablet machines.[5,6]
form to another (e.g., force to voltage). Tablet press
A number of other patents related to press instrumen-
transducers typically measure applied force, turret
tation and control followed from 1973 on ward.[7–15]
speed, or punch position. Because the signals coming
Despite the availability of published designs for
from such transducers are normally in millivolts,
instrumenting rotary presses, much of the early work
they need to be amplified and then converted to
on compaction properties of materials was done on
digital form in order to be processed by a data acqui-
instrumented single punch eccentric presses primarily,
sition system.
due to the relative ease of sensor installation, as well as
availability of punch displacement measurement.[16–18]
In mid-1980s, custom-made press monitoring
systems were described,[19,20] and the first commercial Piezoelectric Gages
instrumentation packages became available, including
both the systems for product development and press Historically, high impedance piezoelectric transducers
control. The first personal computer-based tablet press that employ quartz crystals were used in early stages
monitor was sold in 1987, and the first Microsoft of press instrumentation. When subject to stress, the
Windows-based press instrumentation package in 1995. crystal accumulates electrostatic charge that is directly
A new era of compaction research has begun proportional to the applied force. Both low and
with the introduction of an instrumented compaction (more modern) high impedance piezoelectric gages
simulator[21] in 1976, while a compaction simulator have high-frequency response, but may exhibit signal
patent was issued as recently as 1996.[22] drifting due to charge leakage (approximately 0.04%
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-120012030
3684 Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved.
Tablet Press Instrumentation 3685

decay per second can be seen in modern piezoelectric Once the bridge is balanced, a small perturbation in
transducers). Nowadays, the preferred way of instru- the resistance of any ‘‘leg’’ of the bridge results in a
menting tablet presses is with strain gages. non-zero signal output.
The output of the Wheatstone bridge is normally
expressed in millivolts per volt of excitation per unit
Strain Gages of applied force. For example, sensitivity of 0.2 mV/
V/kN means that applying, say, 10 kN force and
Strain gages are foil, wire, or semiconductor devices 10 V excitation will produce 20 mV output. To utilize
that convert pressure or force into electrical voltage. such output, it usually needs to be amplified several
When a stress is applied to a thin wire, it becomes hundred times to reach units of volts.
longer and thinner. Both factors contribute to Another important function of the bridge is balanc-
increased electrical resistance. If an electrical current ing of temperature effects. Although individual strain
is sent through this wire, it will be affected by the gages are sensitive to temperature fluctuations, Wheat-
changes in the resistance of the conduit. This principle stone bridge arrangement provides for temperature

Tablet–Tablet
is used in strain gage-based transducers. Foil gages, sensitivity compensation, so that the resulting trans-
known for robust application range, useful nominal ducer is no longer changing its output to any signifi-
resistance, and reliable sensitivity control, are most cant degree when it heats up.
commonly used for instrumentation of compression, Because the output of these bridges is in the
precompression, and ejection forces. Semiconductor- range of millivolts, the cables utilized to carry the sig-
based strain gages are inherently more sensitive but nal are normally shielded with a braided or foil-lined
suffer from high electrical noise and temperature sensi- sheath around individual wires. The shield, as a
tivity. Such gages are not commonly used in tablet rule, is connected to the amplifier, but never touches
press instrumentation except for measurement of the actual instrumented equipment (i.e., tablet press).
die-wall pressure and take-off forces. If this rule is violated, a ground loop may generate
electrical noise and present a dangerous electrical
shock hazard.
Wheatstone Bridge

Wheatstone bridge (Fig. 1) is a special arrangement of Gage Factor

strain gages that is used to ensure signal balancing. The
‘‘full’’ bridge is composed of two pairs of resistors in a The gage factor is a specific characteristic of a strain
circle, with two parallel branches used for input and gage. It is calculated as the change in resistance relative
two for signal output. By applying the so-called exci- to unit strain that has caused this change. Strain gages
tation voltage (typically, 10 V DC) to the bridge input that are commonly used for tablet press instrumen-
and changing the resistance of different ‘‘legs’’ of the tation are made from copper–nickel alloy and have a
bridge by adding special resistors, we can make sure gage factor of about 2. In a typical bending beam
that there is a zero output voltage when no load is application (such as compression roll transducer),
applied to the transducer. This is called zero balance. one side of the beam experiences tension while the
other side undergoes compression. By mounting two
gages on each side, the sensitivity of the transducer
can be doubled.
Strain gages have to be bonded to areas of machine
IEX
parts that are most sensitive to applied force. Such
areas can be identified with the use of polarized light
technique that ‘‘points out’’ the stress distribution.
RS1 RS2

Manufacturing a Force Transducer

∆ V to DA
system
RS4 RS3 Usually, instrumentation designs are proprietary and
specific detail drawings are held in fiducial capacity.
Each force transducer is custom designed for a parti-
cular machine part. Overall specifications are taken
Screw from the actual party and/or manufacturer approved
terminals
drawings.
Fig. 1 A typical Wheatstone bridge arrangement of strain Duplicate steel members, such as pins and cams, are
gages. normally made from A2 tool steel in a fully annealed
 3686 Tablet Press Instrumentation

(softened) state. Original machine parts are first Like any properly made transducer, it produces an
annealed, if required. The member is then machined, output signal proportional to the applied load. Unlike
hardened to Rockwell 60–64, tested, and finally, custom-made transducers that require application of
ground to specified dimensions. strain gages directly to press parts, load cells are self-
It is highly advantageous to determine stress contained and can be placed on a press in specially
distribution using polarized light beams identifying the machined cavities to be easily replaced or serviced.
areas of maximum strain yet avoiding areas of uneven As a drawback, load cells generally are less sensitive
strain. or less suited to measure the absolute force than
A typical procedure of transducer manufacturing in custom-made strain gage transducers. Load cells can
a professional gaging lab might be as follows. Foil be used on punch holders in a single station press.
type strain gages are bonded to the steel member utiliz- Another example of load cell use is a die assembly
ing a high-performance 100% solid epoxy system for calibration of existing traducers on a press.
adhesive under controlled heating rate conditions.
After intrabridge wiring is completed using solid
Tablet–Tablet

conductor wiring (to prevent electrical noise), multi- Linear Variable Differential
strand wire cabling with a combination of foil and Transformer
braided insulation is connected to the bridge. Wire
anchoring is achieved utilizing epoxy adhesive and Linear variable differential transformer (LVDT,
protected with a combination of latex-based adhesives Fig. 2) is a device that produces voltage proportional
and/or epoxy resins. Lead wire cabling is protected to the position of a core rod inside a cylinder body.
by Teflon outer shield, as well as inner braided wire It measures displacement or a position of an object
shield. The Teflon to steel joint is sealed with epoxy relative to some predefined zero location. On tablet
but should not be subject to stresses that would cause presses, LVDTs are used to measure punch displace-
the cable to kink or yield in such a way as to expose the ment and in-die thickness. They generally have very
inner braided wire shield. Protective coatings are high precision and accuracy, but there are numerous
then applied and final postcure heating is performed practical concerns regarding improper mounting or
for at least 2 hr at a temperature approximately maintenance of such transducers on tablet presses.
50
C above the transducer normal operating tempera-
ture (approximately 175 or 105
C). A silicon-based
adhesive, such as RTV, is used to fill large cavities Proximity Switch
while maintaining a low modulus of elasticity, prevent-
ing undue influence upon the actual strain measure- Proximity switch (Fig. 3) is a non-contact electromag-
ment. The coating is resilient to moderate mechanical netic pickup device that senses the presence of metal.
abrasion, as well as most solvents, oils, cleaners, etc.
It is not intended for protection against penetrating
sharp objects.
A high-quality connector is then attached to the
cable. The next step is to perform offset zeroing with
fixed, 1% precision, low-temperature coefficient resis-
tors, followed by NIST (National Institute for
Standards and Technology) traceable calibration.
It is worth noting that, in general, the duplicate
members are not made of corrosion-resistant steel,
because high tensile strength and ability to be easily
machined are required. Prior to storage, the surface
should be treated like any high-grade tablet press
punch steel will be treated; a thin coating of oil should
be used after wiping with alcohol.

Load Cells

In some cases, where appropriate, a load cell can be

used in place of bonded strain gages.
A load cell is a strain gage bridge in an enclosure
forming a complete transducer device. Fig. 2 Linear variable differential transformer (LVDT).
Tablet Press Instrumentation 3687

Instrumentation Terms: Definitions

Several important terms may be now defined with

respect to transducers and signal conditioning:

 Full Scale (FS): The total interval over which a

transducer is intended to operate. Also, it can define
the output from transducer when the maximum
load is applied.
 Excitation: The voltage applied to the input
terminals of a strain gage bridge.
Fig. 3 Proximity switch.
 Accuracy: The closeness with which a measure-
ment approaches the true value of a variable being
measured. It defines the error of reading. Good tab-

Tablet–Tablet
On tablet presses, it is widely used to detect the beginning let press transducers have at least 1% accuracy
of a turret revolution, to identify stations and to facilitate (with this level of accuracy, for example, a com-
peak detection in tablet force control applications. pression force transducer with 50 kN FS will pro-
duce at most an error of 500 N).
 Precision: Reproducibility of a measurement, i.e.,
Instrumentation Amplifier how much successive readings of the same fixed
and Signal Conditioning value of a variable differ from one another. If a per-
son is shooting darts, for example, the accuracy is
Signal conditioning involves primarily an amplifier determined by how close to the bull’s eye the darts
that provides the excitation voltage, as well as gain (a have landed, while the precision will be indicated by
factor used in converting millivolt output of the strain how close the darts are to each other.
gage bridge to the volt-based range of the input of the  Resolution: The smallest change in measured
data acquisition device). value that the instrument can detect.
Instrumentation amplifier is different from other  Dynamic range of a transducer: The difference
types of amplifiers in that the signal from each side between the maximal FS level and the lowest detect-
of the transducer bridge is amplified separately and able signal. Measured in decibels (dB), it indicates
then the difference between the two amplified signals the ratio of signal maximum to minimum levels:
is, in turn, amplified. As a result, noise from both sides
of the sensors is reduced.
dB ¼ 10 log10 ðSmax =Smin Þ
Some instrumentation amplifiers also offer filter
functionality. A filter circuit combines resistors and
capacitors that act to block the undesirable frequen- Some press sensors may have a rather narrow
cies. It is a fact, however, that a good transducer dynamic range not necessarily correlated with accu-
should provide a clean signal. Use of analog or digital racy. For example, a very accurate compression roll
filters may cause the loss of some important portions pin designed to measure 50 kN force may not detect
of the signal that one is attempting to measure. Fre- 5 kN signal.
quency filters may cause the measured compression  Calibration: Comparison of transducer outputs at
force peak, for example, to be skewed toward the trail- standard test loads to output of a known standard
ing edge of the peak and can yield a lower than actual at the same load levels. A line representing the best
peak force. fit to data is called a calibration graph.
Because filters distort the signal, they must be  Calibration factor: A load value in engineering
avoided unless absolutely necessary. In many cases, units that a transducer will indicate for each volt
better electronics may make filter use obsolete. For of output, after amplifier gain and balance. Cali-
example, the so-called antialiasing filter that is used bration factor is usually expressed in relation to FS.
to condition a high-frequency signal for a slow sam-  Shunt calibration: A procedure of transducer testing
pling rate is generally not required for tablet press when a resistor with a known value is connected to
applications if modern fast speed data acquisition one leg of the bridge. The output should correspond
devices are used for signal processing. to the voltage specified in the calibration certificate. If
In addition to amplification, excitation, and filter- it does not, something is wrong and the transducer needs
ing, signal-conditioning devices may provide isolation, to be inspected for possible damage or recalibrated.
voltage division, surge protection, and current-to-  Sensitivity: The ratio of a change in measurement
voltage conversion. value to a change in measured variable. For example,
 3688 Tablet Press Instrumentation

if a person ate a 1 lb steak and the bathroom scale part in 216 ¼ 65,536, or approximately 0.0015% of
shows 2 lb increase in the body weight, then the FS (for tablet press applications, such resolution is
scale can be called as insensitive (ratio is far from usually excessive).
unity). Thus, resolution of ADC board limits not only the
 Traceability: The step-by-step transfer process by dynamic range but also the overall system accuracy.
which the transducer calibration can be related to Alternatively, a higher resolution may be required
primary standards. During any calibration process, to retain a certain level of accuracy within a given
transducer is compared to a known standard. dynamic range. For example, a 0.5% accurate trans-
National or international institutions usually pre- ducer with 80 dB dynamic range requires at least
scribe the standards. In the United States, such gov- 12-bit ADC resolution.
erning body is the NIST. Amplifying a low-level signal by 10 or 100 times
 Measurement errors: Any discrepancies between the increases the effective resolution by more than 3 and
measured values and the reported results over the 6 bits, respectively. On the other hand, increasing an
entire FS. Such errors include, but are not limited to: ADC resolution cannot benefit the overall system
Tablet–Tablet

accuracy if other components, such as amplifier or

 Nonlinearity: The maximum deviation of the transducer, have a lower resolution.
calibration points from a regression line (best When an input signal change is smaller than the sys-
fit to the data), expressed as a percentage of tem’s minimum resolution, then that event will go
the rated FS output and measured on increas- undetected. For example, for an FS of 10 V (corre-
ing load only. sponding to, say, a compression transducer output of
 Repeatability: The maximum difference 50 kN), using a 12-bit ADC, any signal that does not
between transducer output readings for repeated exceed 2.44 mV (12.2 N) will not be seen by the system.
applied loads under identical loading and The ADC boards also differ by the effective sam-
environmental conditions. It indicates the ability pling rate range. Sampling rate speed is measured in
of an instrument to give identical results in suc- Hertz (times per second). The signals coming from a
cessive readings. tablet press have a frequency of not more that 100 Hz
 Hysteresis: The maximum difference between (compression events per second). To avoid aliasing (los-
transducer output readings for the same ing resolution of the incoming signal due to slow sam-
applied load. One reading is obtained by pling rate), the sampling rate should be at least twice
increasing the load from zero and the other the highest frequency of the signal. Most ADC boards
reading is obtained by decreasing the load from used for data collection in tableting applications have
the rated FS load. Measurements should be a sampling rate of 5–20 times larger than signal fre-
taken as rapidly as possible to minimize creep. quency. That is why antialiasing filters are not required.
 Return to zero: The difference in two readings:
one, at no load, and the second one, after the
FS load was applied and removed. Computers and Data Acquisition Software

A good transducer is one with the combined (or Overall accuracy of a data acquisition system is
maximum) error of less than 1% of the FS. determined by the worst-case error of all its compo-
nents. One should be aware of the fact that most sys-
tem errors come neither from transducers (0.5%–1%
Analog-to-Digital Converters accuracy) nor from A/D converters (0.025% accuracy)
but from the software analyzing the data (round-off
In order to convey analog output (in volts) from a errors, improper sampling rate, or algorithms).
transducer to a computer, it has to be converted into The speed and capacity of a data acquisition system
a sequence of binary digital numbers. Modern analog- depend on the computer’s processor and hard drive
to-digital converter (ADC) boards are sophisticated specifications. The real-time data from transducers is
high-speed electronic devices that are classified by the streamlined to both the screen (for monitoring) and
input resolution, as well as the range of input voltages the disk (for replay and analysis). Generally speaking,
and sampling rates. ‘‘real-time’’ processing means reporting any change in
Resolution of an ADC board is measured in bits. the phenomena under study as it happens. Interest-
Bit (abbreviation for binary unit) is a unit of infor- ingly, but a high-speed data collection from a tablet
mation equal to one binary decision (such as ‘‘yes or press and a bookkeeping home finance program used
no,’’ ‘‘on or off’’). A 12-bit system provides a resolu- on a monthly basis to balance the checkbook can both
tion of one part in 212 ¼ 4096, or approximately be related to as ‘‘real-time’’ software. The difference is
0.025% of FS. Likewise, 16 bits correspond to one in the time frames. For a tablet press, we are collecting
Tablet Press Instrumentation 3689

data that need to be sampled and processed in milli- upper and lower punch, and moreover, we can com-
seconds (a typical compression event may take 25 msec), pare readings of the compression force from different
while for home bookkeeping once a month will do. tablet presses.
Most vendors supplying transducers, signal con-
ditioning, and computer hardware adhere to practical
standards, e.g., there are some accepted norms for Compression Force Measurement
strain gage factors, combined errors, sensitivity, ADC
resolution, and sampling rate. The difference between On a typical R&D grade compression transducer, a
vendors is apparent when we compare software compression roll pin is machined with incisions made
because there are no universally established standards for placing strain gages (Fig. 4). The actual form of
of user interface. Yet the hardware is ‘‘transparent’’ these cavities constitutes the very art of the transducer
(i.e., invisible) to the end user—day in and day out design that is usually proprietary and is based on the
the user is facing the screen, keyboard, and mouse. know-how of instrumentation vendor.
The ease of software use, bug-free analysis of signals It hast to be noted that there could be an upper or

Tablet–Tablet
coming from transducers, reliability of statistical lower instrumented compression roll pin.
computations, quality of graphs and reports, and vali- The resulting measurement of a compression force
datability of the system—all of the above contribute to is highly correlated with a variety of tablet properties.
the quality of the data acquisition software. As compression increases, so does tablet hardness and
Proper validation tests of a data acquisition system weight (at constant thickness and true density), along
should include calculation of an overall system error with a force required to eject a tablet. Many variables
when the input is known and controlled (e.g., an NIST affect the force of compression: press settings, press
traceable signal generator providing a sinusoidal signal speed, punch length variation, punch wear, and dam-
with a known amplitude and frequency, to simulate age, formulation and excipient properties.
compression events on a press). Comparing the output
(for example, peak heights as reported by the software)
to the known input, the overall system error can be Precompression Force Measurement
reliably established.
Similar to instrumented compression pin, there is a
way to instrument an upper or lower instrumented pre-
TABLET PRESS INSTRUMENTATION FOR compression roll pin. Precompression, if it exists on
PRODUCT AND PROCESS DEVELOPMENT a press, is used for de-aerating and initial tamping
of the powder mass in the die and usually helps to
R&D Grade Instrumentation achieve the desired hardness without capping or
lamination.[30–31] The force of precompression is nor-
In a production environment, typical tablet weight mally a fraction of the compression force.
control mechanism is driven by signals that are coming
from a compression force transducer. Strain gages may
Ejection Force Measurement
be installed on a column connecting the upper and
lower compression wheel assemblies. This transducer
There are many ways to instrument an ejection cam. A
measures what is generally known as ‘‘main com-
preferred arrangement of strain gages (the so-called
pression,’’ i.e., a diluted average of the forces acting
‘‘shear force’’ design) does not require any discontinui-
on the upper and lower punches. Alternatively, a load
ties in the cam surface, and, most importantly, it
cell may be positioned in some convenient location to
provides for a very good linearity of the resulting
register a transmitted compression force. It is measured
signal (Fig. 5).
away from the force application axis, and some force is
lost in the transmission. The resulting measurement
may be adequate for tablet weight control, but, even if
properly calibrated, it does not represent the absolute
value of the compression force.
The R&D grade instrumentation, in contrast,
requires placing the strain gages as close to the punch
tips as possible in a vertical alignment with the direc-
tion of force application.[30] In practice, it means plac-
ing the gages in a compression roll pin, so that the
resulting measurement would reflect the absolute force.
Thus, we can differentiate between the force on the Fig. 4 Compression force transducer.
 3690 Tablet Press Instrumentation

switch). It is installed on the press in order to mark the

beginning of a turret revolution and thus enable sta-
tion identification and speed calculations by system
software.
In addition, a linear arrangement of proximity
switches can mark the beginning and end of a com-
pression event (information that is vital for tablet
weight controllers) and also indicate missing punches.
Other points of instrumentation on a tablet press
are as follows:

 A rotary press pull-up cam can be instrumented to

measure the upper punch pull-up force (the force
required to pull up the upper punch from the die).
Tablet–Tablet

Fig. 5 Instrumented ejection cam. Likewise, the lower punch pull-down force is mea-
sured on a bolt holding the pull-down cam.[34] It
is useful in determining the smoothness of press
operation (extent of lubrication, cleanliness of the
Larger ejection forces may lead to an increased wear
machine, and long batch fatigue buildup).
on tooling and eject cam surface. Ejection force may
 Punch displacement measurements are easily done
also be used to evaluate the effectiveness of lubrication
on a single station press by attaching LVDT to
(of both the press and the product) and punch sticking.
the punch. On a rotary press, such measurements
Sensitivity and linearity of an ejection transducer are
can be done by means of slip ring, telemetry, or
design dependent: shear force designs are always pref-
instrumented punch. Punch displacement profiles
erable over split cam or cantilevered beam designs.
may be used in conjunction with compression force
to estimate work of compression and work of
Take-Off Force Measurement expansion (measure of elasticity). Because capping
tendency increases with the punch penetration
Take-off force is monitored by mounting a strain gage depth, it may be desirable to monitor actual punch
to a cantilever beam on a press feed frame (in front of movement into the die. The shape of a force–
the take-off blade, Fig. 6). It is done to measure adhesion displacement curve is an indication of the relative
of tablets to lower punch face. Such adhesion is indica- elasticity or plasticity of the material; whereas plas-
tive of underlubricated granulation, poor tooling face tic deformation is desirable for stronger tablets,
design, die-wall binding, and tablet capping.[32,33] excess plasticity usually results in tablets that tend
to cap and laminate.[35–37]
 Radial and axial die-wall force measurements also
Speed Measurement and provide an insight into the compaction mechanism
Station ID Determination of the material and may indicate a die-wall binding
(sticking) that is, in effect, a negative pull on lower
Station identification and press speed are usually punch. The radial die-wall pressure due to friction
obtained by means of a revolution counter (proximity is material-specific and is more evenly distributed
inside the die with an addition of a lubricant.[38–44]
Instrumentation of the die presents a technological
challenge because pressure is distributed non-
linearly with respect to tablet position inside the
die and depends on tablet thickness.[45,46]
 In-die temperature can be monitored for heat-
sensitive formulation, such as ibuprofen.[47,48]

Instrumented Punch

Several vendors offer instrumented punch, i.e., a punch

that has strain gages and other instrumentation built-in.
Fig. 6 Instrumented take-off bar. The data are then accumulated or transmitted via
Tablet Press Instrumentation 3691

telemetry to a stationary computer. Such devices are sticking punches, underlubricated dies, and so on.
versatile enough to report compression forces and Finally, instrumentation is widely used for tablet
either punch displacement or acceleration, and, at least weight control.
in theory, they can be easily moved from press to
press.[49,50] However, one should keep in mind that
each instrumented punch is limited to one size and Instrumentation for Formulation
shape of the tooling, and is limited to one station, com- Development
pared to roll pin instrumentation that reports data for
all stations and any tooling. In addition, instrumented Much of the current body of knowledge about com-
punches are either rather cumbersome to install, or else paction properties of pharmaceutical materials came
they report a useless measurement of punch accelera- from instrumented tablet presses.[52–56] Many tablet
tion instead of punch displacement. Attempts to calcu- properties, such as tensile strength (hardness) and
late displacement from acceleration so far could not be porosity can be predicted from force profiles.[57–59]
validated. Work of compaction (a scale-up parameter) can be

Tablet–Tablet
obtained with proper instrumentation.[60,61] Infor-
mation about the plasticity of materials can be derived
Single Punch Press for R&D
from force–time curves.[62–64]
The phenomenon studied with the help of instru-
Single punch eccentric presses are often used in early
mentation is the so-called ‘‘lag time’’ (the time differ-
stages of formulation development because they do
ence between peak of compression and maximum
not require a large amount of powder. Another benefit
punch penetration). The extent of this lag is indicative
is that they allow relatively inexpensive measurements
of compaction mechanisms of the powder being com-
of die forces and punch displacement (there is no
pressed.[65,66]
rotation of die table and therefore no need to use
expensive telemetry methods). This is the primary rea-
son why so much basic research and product develop- Typical waveform
ment were done on eccentric machines.[51] Negative
considerations are that a special tooling is required Let us have a look at the actual waveforms that one
(usually, F tooling), and also that speed of compaction may obtain from an instrumented tablet press (Fig. 7).
is too slow compared to rotary presses. As it will be The black proximity switch trace that marks the begin-
seen later, the speed is a crucial factor in tableting pro- ning of a revolution should be noticed. On this press,
cess and therefore the results obtained on single punch precompression and compression events coincide in
presses do not directly correlate with tablets made on time (they relate to different punches, of course). They
rotary machines. are followed by the ejection and take-off.

Benefits of Press Instrumentation Compactibility profile

Among many benefits of press instrumentation, formu- When the average tablet hardness is plotted against
lation fingerprinting is perhaps the most obvious. the average compression peak force, we get the so-
Compressibility and ejection profiles, as well as dissolu- called compactibility profile that allows us to compare
tion and disintegration curves related to compression different formulations or different processing speeds.
force, are unique for each formulation and can be Referring to Fig. 8, which formulation is better? Well,
used as a batch record. For process optimization, one formulation No. 2 makes harder tablets for the same
can include compression or precompression force and compression force, and this would mean less wear
speed factors in experimental design. Compactibility and tear on the production press is required to achieve
and ejection profiles can be used for excipient and desired hardness. On the other hand, if the hardness
lubricant evaluation. Other useful product develop- tolerance limits are exceptionally narrow, the steeper
ment and optimization tools include response surface, slope of formulation No. 2 may be a detriment.
Heckel, and force–thickness plots.
Scientifically reliable process scale-up cannot be Lubricant profile
achieved without instrumentation data providing
scale-up parameters such as dwell time, density, and A certain quantity of lubricant must be present in the
energy of a tablet. granulation to reduce the friction that occurs at the
In a pilot plant and on the production floor, proper die wall as the tablet is being ejected, as well as to pre-
instrumentation can be used for press troubleshooting, vent sticking of the tablet to the face of the punches.
to warn about tooling irregularities, worn-out cams, Without instrumented ejection cam and take-off bar,
 3692 Tablet Press Instrumentation
Tablet–Tablet

Fig. 7 Typical compression, precompression, and ejection waveforms.

no objective estimate of an optimal lubricant level is Response surface plot

possible, and the ‘‘best’’ formulation is usually the last
one prepared. Formulation and process optimization can be done
The plot shown in Fig. 9 will help a formulator to statistically with the use of experimental design for esti-
determine the optimal amount of lubricant. Obviously, mates of the best processing parameters and excipient
one would try to minimize the ejection force (again, to and lubricant levels. Controllable variables in tableting
reduce wear and tear on the ejection cam of the pro- are mainly the precompression and compression forces
duction press), and yet to avoid the pitfalls of having and tablet press speed, as well as the formulation
too much lubricant in the formulation. component levels. Response variables include the ejec-
Because of the natural association of lubricant tion force, tablet hardness and friability, dissolution
properties with lipophilic materials, formulations con- rate, and drug stability. The purpose of an experi-
taining high levels of lubricant can show retarded dis- mental design is to perform a series of experiments in
solution of the active ingredient and the slow
dissolution rate could adversely affect the in vivo bio-
availability of the drug.
Formulation 1 = 3.9 + 0.305 x
Formulation 2 = 2.2 + 0.486 x
Lubricant study 19

17
Another important use of ejection force transducer
15
Hardness, kP

is the evaluation of lubricants (either different chemi-

cally, or similar lubricants coming from different 13
vendors). 11
As shown in Fig. 10, the preferred lubricant is No. 1 Formulation 1
9
as it results in a smaller ejection force. Early lubricant Formulation 2
studies are a must. When lubricant problems occur 7

later on in the scale-up process, corrective measures 5

8 12 16 20 24 28 32 36
not only require additional materials and development Compression, kN
time, but may also require a supplement of an
approved new drug application (NDA). Fig. 8 Compactibility profile.
Tablet Press Instrumentation 3693

Ejection force vs. compression force at

z := 0.004138x + 0.000034xy − 0.000001x2
different lubricant levels
140

120 1.5% Lubricant

1.0% Lubricant
20 −1.544
Ejection force, lb

100 0.5% Lubricant

−0.479
16
0.587
80
12

Hardness, kP
1.653
2.718
60 8
3.784
4 4.850
40
5.915
0
6.981
20 8.047
500 1500 2500 3500 4500 5500 6500

0
1
1
9.112

50 00
Compression force, lb

0
9

00
45 00
10.178

40 00
0
M

Tablet–Tablet
7

35 00
CC 11.244

30 00
Fig. 9 Lubricant profile.

N
25 0
,m

k
12.309

20 00

n,
g

0
0

15 0

io
3

s
10 0
13.375

es
0
50

pr
0
1

om
14.441

C
order to determine some levels of factors that will Fig. 11 Response surface plot.
allow us to achieve an optimal level of dependent vari-
ables. The experiments should be designed so as to
minimize effort and maximize statistical reliability of If you monitor punch displacement and com-
the results. Published work in this area deals mostly pression force, you can make pretty accurate assess-
with response surface designs that produce a predictor ment of the compactibility of your material. If
polynomial equation for each response variable under LVDTs are attached to both upper and lower punches,
consideration. A multidimensional surface is then it is possible to actually measure in-die thickness of the
searched for the best ranges of factor variables that, tablet at various compression levels. In Fig. 12, one can
when plugged in the equations, result in the optimal see how the tablet thickness rapidly decreases in the
value of the responses. In this plot (Fig. 11), the opti- compression stage and then gains some thickness dur-
mal (highest) hardness is obtained when the com- ing the decompression stage of the tableting cycle. The
pression force is in the range 3000–4000 lb with as degree of this increase in thickness is indicative of
much MCC in the formulation as possible.[67] the elastic recovery as the pressure is removed from
the tablet.
Compactibility study—elastic recovery Elastic recovery and work of compaction were stud-
ied extensively using instrumented tablet presses.[68]
Many powders, especially with viscoelastic compaction
mechanism, such as starch or avicel, exhibit large Compactibility study—Heckel plot
degree of stress relaxation (with time-dependent defor-
mation). In 1961 Heckel[69,70] postulated a linear relationship
between the logarithm of relative porosity (inverse den-
sity) of a powder and the applied pressure. The slope of

Ejection force vs. lubricant level at 3000lb compression

140
Force – thickness plot
120 Lubricant 1 12
Compression force, kN

Lubricant 2
Ejection force, Ib

10
100
8
80
6 UC
60 4

40 2
0
20
0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 6 5 4 3 2 1 0
Lubricant level, % Tablet thickness, mm

Fig. 10 Lubricant study. Fig. 12 Compactibility study—elastic recovery.

 3694 Tablet Press Instrumentation

2.5

2
–In(Porosity)

1.5
In(Porosity)
1

0.5

0
0 50 100 150
Compaction pressure, MPa

Fig. 13 Heckel plot.

Tablet–Tablet

the linear regression is the Heckel constant, a material-

dependent parameter inversely proportional to the Fig. 14 Upper and lower compression force vs. time.
mean yield pressure (the minimum pressure required
to cause plastic deformation of the material undergoing
compression). Large values of the Heckel constant Histogram of Punch Performance
indicate susceptibility to plastic deformation at low
pressures, when the tablet strength depends on the par- Similar irregularities in punch tolerances become even
ticle size of the original powder. The intercept of the more visible on a bar histogram where each bar corre-
line indicates the degree of densification by particle sponds to peak force produced by each punch (Fig. 15).
rearrangement (Fig. 13). As the new revolution arrives, the bar lengths are
adjusted. If a particular bar persists in being taller or
smaller than the rest, it means the presence of a long
or short punch.
TABLET PRESS INSTRUMENTATION FOR
PROCESS TROUBLESHOOTING
Press Monitor—Real-Time Screen
Typical problems in tableting include weight variation,
capping, lamination, tooling irregularities, die binding, On this real-time screen (Fig. 16), one can clearly see
picking, and sticking. Most of such problems can be that there is an abnormal ejection event in the first sta-
detected and/or resolved using proper instrumentation. tion after the proximity switch. This is a clear indi-
cation of a sticking punch or a similar problem. In
addition, the first compression peak is somewhat taller
Upper and Lower Compression than others. This may be caused by a long punch.

Fig. 14 shows a typical set of upper and lower com-

pression profiles. One can see that the lower trace is TABLET PRESS INSTRUMENTATION
smaller than the upper. On a single station press, only FOR PROCESS SCALE-UP
the upper punch is usually moving, and the difference
is caused, mainly, by the friction of the compressed Scale-Up Factors
powder inside the die.
On a rotary press, both punches are moving and One of the main practical questions facing formulators
therefore the friction of the punches inside the turret during development and scale-up is: Will a particular
and the die causes the difference. The lower punch fits formulation sustain the required high rate of com-
the turret with a larger standard clearance compared to pression force application in a production press with-
the upper punch. In addition, the lower punch is never out lamination or capping? Usually, such questions
leaving the die, while the upper punch leaves and enters are never answered with sufficient credibility, especially
the die with each stroke. This results in a compara- when only a small amount of material is available and
tively better alignment and lower friction. The close any trial and error approach may result in costly
fit of the upper punch does not allow it to penetrate mistakes along the scale-up path.[71]
the die smoothly and this can cause the increase in fric- As formulations are moved from small-scale
tion. Thus, an excessive difference between the two research presses to high-speed machines, potential
peaks may indicate an underlubricated die or some scale-up problems can be eliminated by simulation of
punch misalignment problem. production conditions in the formulation development
Tablet Press Instrumentation 3695

Tablet–Tablet
Fig. 15 Histogram of compres-
sion force per punch.

lab. One way to eliminate potential scale-up problems dissolution that may be affected by porosity). How-
is to develop formulations that are very robust with ever, a formulation that was successfully developed
respect to processing conditions. A comprehensive on a single station or small rotary press may not stand
database of excipients detailing their material proper- up to the challenges of scale-up because tablets that
ties may be indispensable for this purpose. However, were meeting all specifications in the lab or clinical
in practical terms, this cannot be achieved without studies may exhibit capping or lamination at higher
some means of testing in production environment speeds.[72,73]
and, because the initial drug substance is usually avail- Compression force magnitude and the rate of force
able in small quantities, some form of simulation is application are the most important variables in tablet-
required on a small scale. Studies of tableting process ing scale-up.
on a class of equipment, generally known as compac-
tion simulators, are designed to facilitate the develop- Force factor
ment of robust formulations. However, simulators
are rarely used to simulate tablet presses for reasons The compression force is the dominant factor of the
that will be explained later. tableting process. It is directly related to tablet hard-
In any process transfer from one tablet press to ness and friability, and is correlated with the phenom-
another, one may aim to preserve mechanical proper- ena of lamination and capping.
ties of a tablet (density, and, by extension, energy It was also shown to have effect on disintegration
used to obtain it) as well as its bioavailability (e.g., times and dissolution profile.

Fig. 16 Real-time screen.

 3696 Tablet Press Instrumentation

Speed factor various excipients, especially viscoelastic materials.[90–94]

There are at least two definitions of dwell time in practice
As the punch speed increases, so does the porosity of today.
tablets and their propensity to capping and lamination.
The tensile strength of compacts tends to decrease with
Functional definitions
faster speeds, especially for plastic and viscoelastic
materials, such as starch, lactose, avicel, ibuprofen, or
Functionally, the effective dwell time (EDT) at 90%
paracetamol. Such materials have the tendency to
level can be defined as the time it taken by the force–
cap or laminate at higher speeds.[74–88]
time curve to traverse the 90% of the peak height. Like-
The notions of dwell time and contact time, to be
wise, the effective contact time (ECT) is the time
discussed in detail later, are the common indicators
between points at 10% of the peak height. The shape
of press speed and the rate of force application.
of the force curve depends, as we know, on the defor-
mation mechanisms of the powder and therefore, all
Force profile factor
Tablet–Tablet

other variables being equal, EDT and ECT will be dif-

ferent for brittle and plastic materials. Although some-
In addition to the level of force and the rate of force
what useful for material characterization, such
application, the shape of the compression force vs. time
variables should not be confused with the universally
curve is of a paramount importance because it directly
accepted definitions of contact time (time of contact)
affects tablet properties such as hardness and friability.
and dwell time (time of immobility).
It is a known fact that the compression part of the
compression cycle (during the ‘‘rise time’’ of the
force–time profile) is 6–15 times more important than Mechanical definitions
the decompression part as a factor contributing to cap-
ping and lamination. On the other hand, reducing the Mechanical Definitions of dwell and contact times dis-
decompression part of the cycle results in the increase regard material properties and concentrate on press
of tablet hardness by reducing the extent of elastic and punch geometry (Fig. 17). Contact time can be
recovery.[89] Alternatively, reducing the compression defined as the time the punch is in contact with the
part of the cycle results in no change of tablet strength compression wheel. Dwell time is defined as the time
for viscoelastic materials and increased hardness for the flat portion of punch head is in contact with
brittle materials. the compression wheel (time at maximum punch dis-
placement, or time when the punch does not move in
vertical direction). In dwell time calculations, the
Other Considerations length of the punch head flat and horizontal compo-
nent of punch speed (as determined by RPM and pitch
Numerous other factors may affect the scale-up pro- circle diameter) are used. In case of a round head tool-
cess. The quality of the measurements, variation in ing, the dwell time, as defined here, is zero. But it
tooling, powder properties, and tablet weight are some should be kept in mind that mechanical definition is
of those factors. given here as a convention, a yardstick, or a common
measure, to compare press speeds for different presses,
 Instrumentation grade and its absolute value is meaningless. A proposed con-
 Measurement of speed vention to quantify linear speed of a press is to use an
 Measurements of mechanical strength
 Tooling variation
 Powder flow variation
 Excipient/raw material variation
 Tablet weight variation

Dwell Time and Contact Time

Matching tablet press speed (rpm) of the research and

production presses has, of course, no meaning, because
of different number of stations and pitch circle diam-
eter. It is vital, therefore, to translate the rpm into
dwell time or contact time.
Many investigators have reported the effect of dwell Fig. 17 Dwell time and contact time—mechanical defini-
times and strain rate sensitivity on the compaction of tions.
Tablet Press Instrumentation 3697

Tablet–Tablet
Fig. 18 Dwell time comparison for
different presses.

Industrial Pharmaceutical Technology (IPT) Type B Dimensional Analysis of Tableting Process

tooling with a known punch head flat as a standard
for press speed comparisons. Dimensional analysis is a method for producing
In what follows, we will use the mechanical, rather dimensionless numbers that completely characterize
than functional, definitions because they serve as an the process. It is widely used in many areas, including
objective material-independent measure of compaction chemical engineering and pharmaceutical industry.[71]
speed. Because all the dimensionless numbers necessary to
describe the process in similar systems must have the
Dwell time comparison for different presses same numerical value,[95] matching such values on dif-
ferent scales is a sure way to success in any scale-up
Comparing the dwell time ranges for a number of operation. This dimensionless space in which the mea-
presses can be a gratifying experience. Here one can surements are presented or measured will make the
see that even for the same brand name, there is a wide process scale invariant.
distribution of ranges. It has to be noted that proper In tableting applications, the process scale-up
scale-up will be possible only when the ranges overlap. involves different speeds of production in what is essen-
Dwell time range may also be used to classify or ident- tially the same unit volume (die cavity in which the
ify tableting equipment. The dwell time ranges vary compaction takes place). Thus, one of the conditions
considerably in various tablet presses. As shown in of the theory of models (similar geometric space) is
Fig. 18, Manesty Betapress is positioned well within met. However, there are still kinematic and dynamic
the range of production speeds of the high-speed parameters that need to be investigated and matched
presses. That is, probably, why this press is often used for any process transfer.
for R&D work. On the other hand, small presses such Scientifically sound approach would be to use the
as Korsch PH106 or Piccola do not even come close to results of the dimensional analysis to model a parti-
benchmark production speeds of 6 msec–15 msec in cular production environment to facilitate the
terms of dwell time. The MCC PressterTM can reach
the production dwell times while making one tablet
at a time.

Dwell time vs. production rate

For a benchmark production speed of 100,000 tablets

per hour (Fig. 19), dwell time distribution follows an
inverse power relationship, which is expected because
dwell time is a reciprocal function of the press speed.
In general, all production presses should be qualified
with respect to dwell and contact times per benchmark
output. Ideally, product development must be done on
a laboratory press that can match (in terms of the dwell
time) the target production output. Fig. 19 Dwell time vs. production rate.
 3698 Tablet Press Instrumentation

scale-up of tableting process, by matching several

major factors, such as compression force and rate of
its application (punch velocity and displacement) in
their dimensionless equivalent form.[96]

TABLET PRESS AND COMPACTION

SIMULATORS

It can be seen that, as a rule, tablets are formulated at

speeds that are very slow compared to production. If
simulation of a production press is required to mini-
mize the scale-up effort, some way of speeding up the
tableting process in development is required. Once
Tablet–Tablet

the linear speed of the punch is attained, the rate of

force application (i.e., the instantaneous change in
compression) should also be matched. This is, of
course, an infinitely more difficult task.

Hydraulic Compaction Simulator

A small number of devices known as compaction simu-

lators exist in the world. Invented more than 20 years
ago, they become popular in basic compaction
research. A wealth of studies have been generated in
the last 20 years.[97–117]
Compaction simulators (Fig. 20) were designed to
Fig. 20 Schematic depiction of a compaction simulator.
mimic the compression cycle of any prescribed shape
by using hydraulic control mechanisms that are driving
a set of two punches (upper and lower) in and out of the
die. All hydraulic compaction simulators are similar in The compression profile (force vs. time) is impossible to
design and construction. A compaction simulator con- calculate theoretically because it has a different shape
sists of several main units: the load frame (column sup- for different materials and tooling.
ports and crossheads with punches), the hydraulic unit Using an instrumented punch to collect force data is
(pumps and actuators that move the crossheads), and cumbersome because it is limited to a particular punch
the control unit (electronic console and computer). size and shape. Recalculation to pressure values is not
Usually, a simulator accepts F tooling only, but can be always adequate. One can, however, monitor and
retrofitted to use standard IPT B tooling. Under com- record force waveform from a properly calibrated
puter control, the hydraulic actuators maintain load, R&D grade compression transducer. Once the pro-
position, and strain associated with each punch. duction press brand, model, speed, and tooling are
The simulation can be achieved by one of the two specified, a waveform can be recorded and then fed into
procedures: matching either the force (load control) or hydraulic simulator.
the movement of punches (position control) at any given This approach is impractical for formulation devel-
moment of time. Thus, when running a simulator, one opment work because one would need to record force
has a choice: to mimic the force/time path (compression waveform on a rotary press using formulation similar
profile) or the motion of the punches (punch displace- to the one being developed. Usually, there is no such
ment curves). It is impossible to mimic both at the same powder available and the actual formulation being
time on any hydraulic compaction simulator. developed is available in limited quantities.

Load control profile Position control profile

Matching the force–time profile of a production tablet Because load control profiles are not practical, users
press is the primary goal of any tablet press simulation. of hydraulic compaction simulators overwhelmingly
However, the rate of force application and the shape of prefer to utilize punch displacement profiles in hope
the resulting signal are not usually known in advance. that, once the punches are forced to move in the same
Tablet Press Instrumentation 3699

pattern as in the production press, the force–time curve empty die. The effect of material resistance to pressure
will follow. For example, a recently built laboratory and elastic recovery is not accounted for in the equa-
compaction simulator does not have a force control tion. The discrepancies between the calculated and real
functionality at all.[118] punch movements are rather striking.[120–122]
There are three possible sources of the punch In addition, it was shown that the lower and upper
position trace: punches may not move synchronously. Moreover,
maximum force does not coincide in time with the
 Prerecorded data minimum punch gap.[123] These and other considera-
 Artificial profiles tions (press deformation, contact time, etc.) make the
 Theoretical profiles effort of simulating a production press on a hydraulic
compaction simulator rather impractical. That is why,
To obtain a position control profile from any tablet to quote from a paper by Muller and Augsburger,[124]
press, one has to record the movement of the punches ‘‘Although compaction simulator have been designed
using LVDT. Besides the technological challenge that to mimic the displacement time behavior of any tablet

Tablet–Tablet
this objective may present, the punch movements on press, they rarely have been used in that fashion.’’
a press depend on many factors, including brand name Literature sources reporting the use of compaction
and model of the press, speed of the turret, shape of simulators in simulating actual tablet presses are rather
punches and die, size and shape of the tablet, and most scarce. Some studies suggest that, for whatever reason,
importantly, compaction properties of the powder. The tablets made on a Manesty Betapress were significantly
problem with this is that data from production presses softer than those made on a compaction simulator
are inevitably obtained using material other than the using the theoretical Betapress punch displacement
one being developed. It is a vicious circle: the profiles profiles.[120,125]
before developing a formulation and the formulation To summarize, one can say that hydraulic compac-
before obtaining punch displacement profiles are needed. tion simulators are ideally suited for basic compaction
Many compaction studies were done on compaction research but are not very practical for simulation of
simulators using artificial punch displacement profiles, production presses.
for example, the so-called ‘‘single-ended’’ profile, i.e.,
when the lower punch is stationary (like in a single
station press). Other studies were using a ‘‘saw tooth’’ Tablet Press Replicator: The PressterTM
profile, i.e., a constant speed profile where punch dis-
placement speed is constant at any given time interval Recently, a new type of machine was introduced to
under load. It is obvious that such profiles have noth- mimic production presses on a small scale. Known
ing to do with simulation, although they provide a under a brand name of ‘‘Presster’’—kind of an
degree of uniformity for basic compaction studies. agglomeration of the words ‘‘press,’’ ‘‘presto,’’ and
The very name ‘‘compaction simulator’’ is a misnomer ‘‘tester’’—this machine can be classified as a mechan-
as is acknowledged by a number of researchers in the ical compaction simulator.
field. The machine is best described as a compaction The PressterTM is a high-speed single station press
research system because it is well suited for the basic that is also a tablet press simulator (Fig. 21). It was
compaction studies (densification and bonding proper- designed to simulate production presses without any
ties of materials). use of hydraulic controls, and, consequently, there is
To simulate tablet presses, compaction simulator no need to feed in any artificial, theoretical, or prere-
users most frequently employ the theoretical position corded punch displacement profiles. Built around a lin-
control profiles. Theoretical path is calculated from ear carriage that moves a set of punches and a die
the geometry of the press and punches, using the radius between two compression rolls, it can mimic press
of the compression roll, the radius of the curvature of geometry by matching the compression wheels match
the punch head rim, the radius of the ‘‘pitch circle’’ press speed using a variable speed motor drive match
(distance between turret and punch axes), and the tur- tablet weight and thickness by adjusting depth of fill
ret angular velocity.[119] and the distance between the rolls match tooling by
The resulting sinusoidal equation is used in order to installing standard IPT or any special tooling.
‘‘simulate’’ punch movement in a tablet press. Thus, using mechanical similarity, all of the scale-up
In practice, theoretical and actual punch displace- fact are matched, namely, the compression force,
ment profiles on a rotary press have very little in the speed, and the shape of the force profile. To use
common because the theoretical profile does not Presster, first a production press to be simulate should
account for several mechanical factors, such as punch be selected, and the compression wheels with a match-
head flat. Moreover, the punch movement equation ing diameter should be installed. Then, production
was derived for a punch moving in and out of an speed should be selected in terms of tablets per hour,
 3700 Tablet Press Instrumentation
Tablet–Tablet

Fig. 21 Tablet press replicator: The PressterTM.

RPM, or dwell time. The PressterTM will mimic the transducer and several proximity switches for station
selected production press speed and compression force identification and pinpointing the compression event.
profile and will allow us to make one tablet at a time. Tablet weight controller can be just one, albeit a major,
As a high-speed single station press, mechanical unit of a larger press automation system. Press auto-
compaction simulator will be able to plot compres- mation system may include:
sibility profiles, Heckel graphs, calculate work of compac-
tion, and virtually any other imaginable variable that is of  Tablet weight control
interest to formulators. Tensile strength of tablets made  Material handling interface
on a Betapress and The PressterTM was similar.[126]  Feeding system
Precompression and ejection steps of the tableting  Collection system
cycle can be included in simulation. Some current lim-  Packaging system
itations of Presster should be pointed out: It will  Supervisory control (SCADA) station
neither follow any artificial punch movement profile,
nor will it address, at least in its present implemen- The latter can be a computer positioned on the
tation, the issues of feeding and die fill at high speeds, supervisor’s desk that monitors the performance of
or speed-related temperature fluctuations. each press on the production floor, with timely status
report for each batch.
There are many reasons why it is imperative to con-
PRESS INSTRUMENTATION AND CONTROL trol tablet weight variation:
ON THE PRODUCTION FLOOR
 Production costs are lowered because there is less
Tablet Weight Control and Tablet waste
Force Control  Productivity is increased because there is a better
equipment utilization
To keep tablet weight within the prescribed tolerance lim-  Batch-to-batch variability is minimized for obvious
its, the required instrumentation includes compression reasons
Tablet Press Instrumentation 3701

When the cost of each out-of-spec wasted tablet is Control algorithms

significant, the savings produced by weight control
quickly add up. Product quality is improved when One-point control is a description of a control when
there is less of a chance to get an out-of-spec tablet any deviation of the compression force from a preset
into the acceptable batch. Last, but not least, is the target level results in a corrective action of the dosing
improved safety because the automated process requires cam. This action can be proportional (P) to the devia-
less human intervention. tion from the target, or it could be correlated with
the integral (I) of the deviation over some time, or it
could be tied in with the rate of change (D, for deriva-
Control mechanisms tive) of the compression force. This would correspond
to proportional, integral, or derivative control types,
For constant tablet thickness, in a small target area of respectively. There also can be a combination of the
tablet weight, compression force is directly propor- control types. Thus, one can have P, I, P þ I,
tional to tablet weight. Force control systems maintain P þ D, and P þ I þ D, control algorithms.

Tablet–Tablet
compression force within prescribed limits by adjusting Alternatively, a two-point control is enacted when
the depth of fill cam. The limits are established empiri- the compression force is outside an acceptable band
cally for each formulation recipe. outlined by the upper and lower tolerance limits. Thus,
Alternatively, a weight control system would require there are separate control limits, rejection limits, alarm
an expressed correlation between force and tablet limits, and shutdown limits, and no respective action is
weight. A few tablets at different force levels can be taken when the signal is inside these limits.
made to correlate the resulting tablet weights with Strictly speaking, one-point control can be viewed
the force values. Next, one can express tablet weight as a special case of the two-point control when the
tolerance limits in terms of the compression force. bandwidth of the control limits is tightened to
The weight control system can adjust the dosing to approach zero.
keep the tablet weight within the desired limits. For
this control theory to work consistently, one needs to
recalibrate force–weight relationship periodically, as Control Systems
the powder properties and tablet mechanical condition
can vary in time. Original equipment manufacturer (OEM)
Control functions include alarm and shutdown control systems
(when any individual force peak or revolution average
exceeds preset limits), force or weight control (usually Almost each tablet press manufacturer offers a system
done on revolution average only), and rejection of that is designed to control the press. Some of these sys-
out-of-spec tablets. With a mechanical gate, usually tems are very sophisticated devices that monitor and
several tablets are rejected at a time, with the bad tablet control a vast array of tableting functions. If, however,
being rejected along with the adjacent good tablets. there are several brand names on the production floor,
With a fast pneumatic–air stream rejection mechanism, any standards in the control system implementation
the control system can pinpoint and reject one bad for different manufacturers should not be expected.
tablet only. Likewise, software interfaces exhibit quite a range of
user-friendliness.
Control criteria In one brand name press, tablet weight control is
achieved by regulating the dosing cam based on powder
Control decisions are usually done as follows: bed thickness in the precompression cycle. This clever
approach is possible because precompression force is
1. Adjust depth of fill cam if revolution average is kept relatively constant by means of pneumatic com-
outside some redefined acceptable limits. pensating mechanism. Under these conditions, tablet
2. Reject each tablet that exceeds individual limits weight is directly proportional to thickness. The sub-
or if the press speed slows down beyond some sequent compression cycle can be done to constant
level. thickness, like on any other press.
3. Sound an alarm if the same station repeatedly
produces bad tablets or if revolution average Generic control systems
exceeds alarm limits.
4. Shut down the press if any tablet is made with An alternative to OEM control systems is generic con-
a force exceeding the shutdown limits, or if trollers that may offer plug-in compatibility with the
revolution average is outside the shutdown brand name controllers and may provide a degree of
limits. standardization.
 3702 Tablet Press Instrumentation
Tablet–Tablet

Fig. 22 Recipe example.

However, such generic controllers may lack the degree when they need to change recipes, etc. Some examples
of sophistication and versatility of the controllers that are of displays, available to operators of instrumented pro-
made by the press manufacturers. One thing to keep in duction presses equipped with controllers, follow.
mind is that not all control systems are created equal,
but all control systems use the same principles.
A decent tablet weight control system should be Recipe example
based on product recipes, provide instant display of
compression force distribution, control charts and In Fig. 22, an example of a typical control recipe with
batch reports on demand, archiving data for sub- dosing cam adjustment, rejection, alarm, and shut-
sequent analysis or documentation, give some measure down limits expressed as a percent deviation from
of standardization, be fully compliant with current vali- the target tablet weight is shown. The software then
dation requirements, and provide a multilevel security, converts all values into the corresponding compression
e.g., password protection for operator and supervisor force levels for control purposes.

Fig. 23 Statistical process control

X-bar chart.
Tablet Press Instrumentation 3703

Bar histogram 7. Holm, J.P., Arming Control for Servo-Adjusted Tablet

Compression Machines. US Patent 3,734,663, 1973.
8. Hooker, D.B. Potted Strain Gage. US Patent 3,791,205,
This chart is similar to one used in tablet press moni- 1974.
toring for press troubleshooting (Fig. 15). It is vital 9. Williams, J.J. Applied Instrumentation Providing Tablet-
for any production press operator. ing Compression Force. US Patent 4,016,744, 1977.
10. Williams, J.J. Force Measurement and Analysis Parti-
cularly Relating to Rotary Tablet Presses. US Patent
Control chart 4,030,868, 1977.
11. Stiel, D.M.; Browski, C.R.; Neugroschel, E.; Williams, J.J.
Tablet Press Related Instrumentation for Use in Develop-
Control chart is a simple graph of peak compression ment and Control of Formulations of Pharmaceutical
force vs. time. Each point on the chart corresponds Granulations. US Patent 4,100,598, 1978.
12. Williams, J.J. Force Measurement and Analysis Parti-
to the average of N tablets made, with N ranging from cularly Relating to Rotary Tablet Presses. US Patent
1 to several revolutions. The horizontal lines would 4,099,239, 1978.
indicate the control limits. 13. Stiel, D.M. Method Invoking Tableting Compression Force
Control for Optimizing Tableted Formulation Parameters.

Tablet–Tablet
US Patent 4,121,289, 1978.
Statistical process control chart 14. Breen, D.T., et al. Tablet Press Controller and Method.
US Patent 4,570,229, 1986.
15. Lewis, D.A., Pharmaceutical Tablet Press Control Mech-
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 3706 Tablet Press Instrumentation

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Tablet–Tablet
Tablet Testing
Saeed A. Qureshi
Health Products and Food Branch, Health Canada, Sir F.G. Banting Research Centre,
Ottawa, Ontario, Canada

INTRODUCTION 3. Establishing stability profiles to achieve shelf

life.
A tablet is perhaps the oldest and most common phar-

Tablet–Tablet
maceutical dosage form. Its popularity is due to its This article provides an overview of the current
convenience in the administration of a drug without techniques used for this purpose and discussion of
the help or supervision of a health care practitioner, the generally accepted standards. Using conventional-
thus providing patients’ freedom and a very cost- release products as a model product and North
effective means of providing a reproducible medication. American standards/specifications, logistics of conduct-
A tablet seldom consists of only the active ingredient. In ing testing and the general practices and instrumen-
fact, a tablet represents a mixture of one or more active tation used are described. Relevant references are also
ingredient(s) with a number of ‘‘inactive’’ ingredients or provided[1–4] for further and specific details of testing
excipients. There are many reasons for formulating a products and possible differences in national and
tablet product with excipients, ranging from manage- regional guidelines.
ment of small dosage amounts of active ingredient to
esthetic reasons of color and shape of a product.
However, the most fundamental and critical objective TESTING OF NATURE OF THE TABLET
of a tablet product is to provide/deliver the active PRODUCTS
ingredient accurately and reproducibly. Therefore, from
this perspective, a tablet is now commonly considered In this category, one seeks to establish whether the
as a drug delivery device. There has also been an tablets are within specifications, for example, the nat-
increased emphasis in developing tablets that provide, ure of the active ingredient (identification), expected
unlike the conventional-release tablets that are of amount (assay), purity (related compounds), and uni-
fast disintegrating/release characteristics, controlled formity of the amount of drug from tablet to tablet
disintegration/release process of the active ingredient. (uniformity of dosage units). Commonly these testing
These tablet products are therefore known by different procedures are described in pharmacopeias under a
names, such as slow-, extended-, controlled-, sustained-, specific name, for example, the names given in parent-
or delayed-release tablets to reflect their drug release heses are referred to as the USP (U.S. Pharmacopeia)
characteristics. These modified drug release products terminology. In addition to these tests, some other tests
provide further convenience to patients by reduced such as friability, hardness, disintegration, etc. are also
frequency of drug administration, e.g., once a day conducted and are described below. Although a num-
instead of three doses per day, thus increasing chance ber of procedures could be described for individual
of compliance as well. However, for establishing the tests, most emphasis will be given to procedures
quality of a tablet product, the fundamentals remain described in the pharmacopeias because these are
the same, i.e., to ascertain that the product delivers usually relatively simpler to conduct and are generally
the intended active ingredient in an accurate and recognized around the world.
reproducible manner. Therefore, tablet testing can be
broadly divided into three aspects or categories:
Identification

1. Confirmation of the nature of the active ingredi- The first and foremost important test in tablet testing is
ent and the product (identity, quantity, impuri- to establish that the tablets contain the labeled active
ties, integrity, etc.). ingredient. For this purpose, usually a fixed number
2. Establishing pharmaceutical availability of the of tablets, e.g., 10–20, are ground and extracted with
active moiety both in vitro and in vivo in appropriate solvent extraction. The extract, with or
humans and, if required, also in animals. without a concentration step, is usually chromatographed
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-120012013
Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved. 3707
 3708 Tablet Testing

along with an authentic standard solution. The iden- Impurities

tity of drug is confirmed based on similarity of ultra-
violet (UV) spectrum and/or retention times using The impurities present in a tablet product may fall
chromatographic analysis. This test is generally quali- under different categories, such as: foreign substances
tative in nature. More sophisticated techniques such that are introduced by contamination or adulteration;
as chromatographic techniques with or without cou- toxic impurities that have significant undesirable bio-
pling with mass-spectrometry may be used. However, logical activity, even as a minor component, and
for routine quality control purposes, the simpler tech- require individual identification and quantitation by
niques such as thin layer chromatography (TLC) or specific tests; concomitant components that are charac-
high-performance liquid chromatography (HPLC) teristic of many bulk pharmaceutical preparations, but
with UV detection are mostly employed. are not considered as impurities, for example, optical
and geometrical isomers; ordinary impurities are those
that are innocuous by virtue of having no significant
biological activity in the amount present. These impu-
Tablet–Tablet

Assay rities may arise out of the synthesis, preparation, and/

or degradation of the product.
One may assume that this test as a quantitative version Generally, most of the impurity profile is established
of the identification test. Again, 10–20 tablets are from bulk pharmaceutical material (the raw material).
ground and the active ingredient is dissolved or extracted However, from the finished product perspective, pro-
in a suitable solvent using the described procedure. ducts are only analyzed to the extent of toxic impuri-
The concentration of the extracted solution is determ- ties. In this case, usually it must be established that
ined using a specific and validated spectroscopic or product is either free from the specific impurities or
chromatographic method against a solution of reference the impurities are within acceptable specifications.
standard. These results are reported as percent of
expected/labeled value. Although the specifications for
assay results differ from product to product, generally
Friability
the expected range for individual active ingredient is to
be within 90%–110% of the labeled amount.
This test is intended to determine, under defined con-
ditions, the friability of uncoated tablets, the phenom-
enon whereby tablet surfaces are damaged and/or
Uniformity of Dosage Units show evidence of lamination or breakage when sub-
jected to mechanical shock or attrition. Commercially
This test is conducted to establish consistency in the available apparatuses known as friabilators are used
content of active ingredient from tablet to tablet. There for the test. Basically, it consists of a drum with diam-
are generally two approaches taken in establishing this: eter between 283 mm and 291 mm and having width of
weight variation or content uniformity. 36 mm–40 mm, made of transparent plastic material
If the active ingredient represents not less than 50% (Fig. 1). The drum is attached to the horizontal axis
weight of the tablet and greater than 50 mg, then one of a device that rotates at 25
1 rpm. The tablets are
may establish uniformity of dosage units using the tumbled at each turn of the drum by a curve projection
weight variation method. A sample of 10 tablets are with an inside radius of 75.5 mm–85.5 mm that extends
weighed individually and results of these weighing
are recorded.
In the case of the content uniformity approach, a on
Rotati
sample of 10 tablets are individually analyzed using
the analytical method described under the assay pro-
cedure. It is mandatory to use content uniformity for
tablets with less than 50 mg of active ingredient and/
or representing less than 50% total mass of the tablets.
The content uniformity approach is preferred over the
weight variation approach as it more precisely reflects
the variation of the active ingredient from tablet to Tablets
tablet.
The required specification for this test is that uni-
formity of dosage unit should be within a range of
85%–115% with a relative standard deviation of less
than or equal to 6%. Fig. 1 Schematic of friability testing apparatus.
Tablet Testing 3709

from middle of the drum to outer wall. Thus, at each

turn, the tablets roll or slide and fall onto the drum
wall or onto each other. Usually, a sample of 10 tablets
are tested at a time, unless tablet weight is 0.65 g or
less, where 20 tablets are tested. After 100 turns, the
tablet samples are evaluated by weighing. If the
reduction in the total mass of the tablets is more than
1%, the tablets fail the friability test. Generally, the test
is done once. If cracked, cleaved, or broken tablets are
obvious, then the sample also fails the test.

Hardness Testing

Tablet–Tablet
A tablet requires a certain amount of mechanical
strength to withstand the shocks of handling in its
manufacturing, packing, shipping, and dispensing. As
discussed before, hardness and friability are most com-
mon measures used to evaluate tablet strength. The
need for testing hardness or crushing strength, in
addition to friability, may be explained with an ana-
logy that friability determines how fragile a tablet is.
If a tablet is more fragile than expected, then the Fig. 2 Schematic of a tablet disintegration apparatus.
friability test will detect its substandard quality. How-
ever, on the other hand, if the tablets are more robust
than desired, a friability test would not detect this
apparatus. A schematic representation of the appa-
deficiency. It is the tablet hardness test that will detect
ratus is shown in Fig. 2. The apparatus employs a
the deficiency.
basket of six tubes with a base of metal sieve. A tablet
The most widely used apparatus to measure tablet
is placed in each tube and is held in place by a plastic
hardness is the Schleuniger apparatus. This, and other
weight. The six-tube assembly, containing six tablets, is
newer electrically operated test equipment, eliminates
suspended using a hanger with a mechanism of vertical
the operator variability inherent in the measurement
motion at a fixed speed. While hanging the six-tube
using older apparatuses.[5]
assembly on the hanger, the assembly is moved in
Generally, the force required to break a tablet may
vertical motion in water or a buffer solution. The time
be expressed in either kilograms or pounds. It is of
for disintegration of each tablet is recorded and should
critical importance to note that because as results will
meet the required time specification.
vary with the specific make and type of the test appa-
ratus used, direct comparison of results obtained on
different types of apparatuses cannot be made. Thus,
the same instrument must be used consistently
PHARMACEUTICAL AVAILABILITY
throughout a particular study.
Checking the quality of drug products using chemical
Disintegration Test analyses, as described above, ensures that the product
is of acceptable quality in its contents. However, this
A disintegration test is a test to establish how fast a does not guarantee that the active ingredient in the
tablet disintegrates into aggregates and/or finer parti- product will be released in an acceptable manner.
cles. The test assumes that if product disintegrates There are a number of factors, such as particle size,
within a short period of time, such as within 5 min, crystalline form, and compression during manufactur-
then the drug would be released as expected and one ing that, individually or collectively, can severely
should not anticipate a problem in the quality of a impact the release of drug from a product thus affect-
drug product. Although this test is in use for some pro- ing its efficacy. That is why drug release characteristics
ducts in pharmacopeias, its use is generally diminishing must be evaluated and established. The quality assur-
in favor of drug dissolution testing, which is described ance around this concept represents one of the major
later in this article. components of product development and later for
When required, the test is conducted using a spe- quality assurance purposes. The gold standard to
cially designed instrument known as disintegration establish drug release characteristics of a product is
 3710 Tablet Testing

based on an in vivo study, i.e., testing bioavailability therefore, the absorption phase dominates, while at
(BA) of the drugs in humans. later times when all the drug is absorbed, it is the elim-
Furthermore, in reality, changes in product formu- ination process that dominates. Although exceptions
lation and manufacturing are anticipated and in many exist, a typical observed blood drug concentration pro-
cases are unavoidable, for example, if there is a change file after dosing is shown in Fig. 3. From the analytical
in ingredient from the supplier, or change in a manu- chemistry perspective, commonly it is not the blood,
facturing component or facility itself. However, such but the extracted plasma or serum that is analyzed
changes create concern about the impact on the drug and is used to reflect drug levels in blood.
release profile and thus the safety and efficacy of the The time to reach the maximum plasma drug con-
product. The only ‘‘gold standard’’ analytical test is centration (Cmax) is termed as Tmax. The slope reflect-
to conduct a BA study to establish the similarity of ing the increase of drug levels in the initial plasma
the new product to the earlier (or the reference) drug profile represents rate of appearance/delivery of
version. However, conducting such BA studies is often drug to the systemic circulation. Depending on the nat-
one of the most expensive and time-consuming pro- ure of the product, the rate of delivery could be critical.
Tablet–Tablet

cesses in the manufacturing of a product. In addition The slope reflecting the decrease of drug levels in the
to the cost and time considerations, ethical concerns terminal phase of plasma drug concentration repre-
also limit the conduct of these studies in humans. sents rate of elimination of drug from the systemic cir-
Thus, because of cost, time, and ethical considera- culation. If one were to draw the curve shown in Fig. 3
tions, it is not always possible to conduct drug release on a semilogarithmic scale, i.e., time vs. log(plasma
studies in humans, and an in vitro drug release evalu- drug concentration), then the terminal portion of the
ation test is the most desirable alternative. For this curve becomes a straight line, as the elimination rate
purpose, a dissolution test for tablet products has been is usually exponential. This rate of elimination is repre-
developed and has become a tool for both product sented by a parameter ke and is commonly known as
development and quality assurance. The test is con- elimination rate constant. These parameters, collec-
ducted routinely at every stage of drug product tively or individually determine the rate of absorp-
development, during the manufacturing and postma- tion/appearance of drug into systemic circulation.
nufacturing stages. A relatively detailed description Another important parameter is the area under the
of the concepts concerning pharmaceutical availability plasma concentration–time curve (AUC). It reflects the
is presented here along with the issues and difficulties extent of drug absorption. From a product quality
of the current approaches so that the reader may be aspect, knowledge of the described pharmacokinetic
aware of the directions of future development. parameters is important. Utilizing these parameters,
one would establish drug release characteristics of a
tablet in vivo.
In Vivo Drug Release Assessment

To assess release characteristics, one would require to Calculation of pharmacokinetic parameters

determine BA and/or bioequivalence (BE) characteris-
tics of a product. Both BA and BE areas are subdisci- Obviously Cmax and Tmax are observational and
plines of pharmacokinetic studies. Therefore, one determined directly from experimental results or the
would require to have some familiarity with the basic curve. Generally, AUC is determined using the trap-
principle of pharmacokinetics. For the purpose of this ezoidal rule, as the arithmetic sum of average plasma
article, the necessary pharmacokinetic concepts are concentration at two sampling times divided by the
described below. For further details see Refs.[6,7].

Bioavailability Studies

When a drug is absorbed from the gastrointestinal (GI)

tract into the blood stream (systemic circulation), it is
distributed throughout the body that results in reach-
ing at the site(s) of its action. Concurrent to distri-
bution, drug elimination from the blood due to
metabolism or excretion to urine commences. There-
fore, at any given time, drug levels in the blood are
the net result of this absorption and elimination pro-
cesses. In the initial phase, the rate and extent of drug
absorption is greater than that for the elimination; Fig. 3 Plasma drug concentration–time profile.
Tablet Testing 3711

differences in sampling times, i.e., (C2 þ C1)/ the drug, based on the area under the curve, should be
2(t2  t1), up to the last measured plasma concen- accounted for.
tration. Such an area under the curve is termed as The plasma samples are stored for later analysis.
AUC0–t. However, as the drug is still in the body and The samples are analyzed using validated analytical
exponentially eliminated after the last determined con- methods.[9] The most commonly used methods are
centration, the extent of drug remaining is determined chromatographic, i.e., HPLC or gas chromatography.
by a formula AUCt1 ¼ Ct/ke. Thus, the total These methods, including the storage conditions, must
AUCTotal ¼ AUC0–t þ AUCt–1. be validated so that accurate and precise results are
To establish drug release characteristics of a pro- assured. Pharmacokinetic parameters from the plasma
duct such as a tablet, one has to determine BA of the drug release profiles are determined for individual
drug. Bioavailability is defined as a measure, relative volunteers. The average values of these parameters
to some standard, of the rate and amount of drug that reflect the BA of the product.
reaches the systemic circulation. As Cmax, Tmax, and
AUC are reflections of availability of drug in plasma,

Tablet–Tablet
these parameters are used for determining BA of pro- Bioequivalence Studies
ducts. By comparing the derived pharmacokinetics
from a BA study, similarity or dissimilarity of drug A BE study is not as such an experimental study, but a
release in vivo is established. form of reporting for a comparative BA study. In a BE
The concept of BA is very critical from the drug study, a BA study is conducted to compare the results
quality perspective. Although, as stated above, chemi- between reference and test products. A test product
cal and physical tests are critical for testing the quality may be a product after a manufacturing or an ingredi-
of tablet products, from the safety and efficacy per- ent change or a generic product. In fact, the whole gen-
spective of drug products, it is the BA studies that eric industry is based on bringing products to the
are necessary. Even when content and quality of the market based on conducting BE studies between inno-
tablets are within specifications, but release of drug in vators and their products. The study has to be conduc-
humans is not as expected, the substandard quality of ted following a statistically valid approach and design,
the products could be dangerous to the health of its users. usually a crossover design, e.g., in one period, one-half
Therefore, BA studies are conducted at many stages of of the volunteers are given one set of dosage form and
tablet product development and manufacturing, in parti- in the second period, the order is reversed.
cular, product development and receiving permit for mar- Once the experimental part of a comparative BA study
keting. A sample protocol for conducting BA studies for a is completed and respective pharmacokinetic parameters
tablet product testing is described below.[8] are derived and compared, the products are declared
A comparative BA study is usually conducted in bioequivalent when they meet the set and expected speci-
healthy humans, usually involving 12–30 volunteers fications for the parameters. The requirements and para-
depending upon the expected variability in the derived meters and their specifications may vary from country to
pharmacokinetic parameters. To minimize variability country. However, the most common standard followed
in derived parameters, the healthy volunteers should is that of the U.S. Food and Drug Administration
have certain characteristics such as age between 18 (FDA). In this case, a 90% confidence interval of
and 50 years, within normal weight range, and free the ratios of the log-transformed values of parameters
from disease or any medication. The study protocol (Cmax and AUC) should fall within the range 80–125.
has to be approved by the institutional ethics committee. Therefore, if one would like to establish impact of
Following an overnight fast, a dose is administered any change on pharmaceutical availability from a pro-
to the subjects with a controlled amount of fluid duct or to develop a new formulation or product in
(250 ml of water). At a prescribed time, following the comparison with an old product, these BA/BE studies
dose, a standardized meal is provided to each volun- remain the standard and are the requirements for
teer, again to standardize conditions so that results showing the safety and efficacy of the altered/new pro-
from any of the volunteers would not affect adversely. duct. However, from the routine quality control pro-
Following the dosage administration, blood samples spective as well establishing quality of a product after
(10–15) are withdrawn from the volunteers. The idea some minor changes in formulation or manufacturing
behind selecting the sampling times is that one would processes, the justification for BA/BE studies on econ-
be able to accurately determine the Cmax. Therefore, omic and ethical reasons becomes difficult. Therefore,
generally more frequent sampling times are needed at often only in vitro drug release testing is conducted.
the lower part of the curve. Similarly, the terminal In the following sections, circumstances for the use of
phase should have a sufficient number of sample times in vitro and in vivo methods are provided. However,
to establish the elimination rate constant. The duration a brief description of the current practice of dissolution
of sampling should be such that most (at least 80%) of testing is first given.
 3712 Tablet Testing

In Vitro Drug Release Assessment types of apparatuses based on different types of mixing
approaches are available commercially and have com-
Drug dissolution testing is a procedure used to evalu- pendial recognition. These apparatuses are known as:
ate drug release characteristics of solid oral products 1) paddle; 2) basket; 3) flow-through; and 4) recipro-
such as tablets. As in this article, it is also referred to cating cylinder. For a detailed description and specifi-
synonymously as in vitro drug release assessment as cations of these apparatuses, readers should refer to
opposed to in vivo drug release, which is commonly any of the pharmacopeias such as the USP.[12] A brief
assessed in humans with studies using blood/plasma description of the apparatuses and stirring mechanism
concentration profile information from BA or BE stu- is given in the following sections.
dies. The rationale behind conducting dissolution test-
ing is that if a drug is to be absorbed from the GI tract, Paddle Apparatus. In this case, the agitation is
it usually has to dissolve. Therefore, for a drug to be achieved by an inverted T-shape stirrer. The product
absorbed, it has to be released from the product and is placed in the medium that is contained in a glass bea-
dissolved in the GI fluid. Thus, a dissolution test is ker with round-shaped bottom (Fig. 4). The stirring is
Tablet–Tablet

an established analytical test to assess the qualities of achieved by rotating the spindle commonly between
a drug product, based on its rate and extent of dissol- 50 rpm and 100 rpm. At specific times, samples are
ution, i.e., release characteristics. withdrawn and the percent of the drug dissolved is
Commonly, dissolution tests are conducted with determined using any of the conventional analytical
apparatuses comprising two major determinants that methods such as UV or liquid chromatography.
can be varied: 1) dissolution medium (nature and
volume); and 2) a stirring and mixing mechanism. Basket Apparatus. This method is very similar to the
one described above except that the inverted T-shape
Choice of dissolution media stirrer is replaced with a cylindrical metallic-wire bas-
ket (Fig. 4). The tablet product is placed in the basket
As a dissolution test is conducted to simulate drug that is attached to the spindle, which provides rotation
release in the human GI tract, the generally recom- to the basket. The dissolved drug comes out of the bas-
mended media are based on aqueous buffers in the ket and mixes with the bulk medium. In this apparatus,
pH range of 1–8. Commonly used media are HCl it is possible that when the product is disintegrated and
(0.01–0.1 N) to simulate gastric fluid and phosphate particles drop and settle at the bottom of the vessel
or acetate buffer in the range of pH 4.0–6.8 to rep- without dissolving thus providing limited dissolution.
resent intestinal fluids. Recently, suggestions[10,11] have Therefore, one may anticipate erratic dissolution
also been made for other relevant dissolution media results using such an apparatus, at least for some pro-
reflecting composition, volume, flow rates, and mixing ducts.
patterns of the fluids in the GI tract. Volumes are
usually in the range of 250 ml–1000 ml, however, the
use of smaller and larger volumes are becoming com- Flow-Through Apparatus. In this type of apparatus
mon as well. A critical consideration in the choice of (Fig. 5), no stirrer is present. The product is placed
a dissolution medium volume is that drug should dis- in the path of a flowing stream of dissolution medium.
solve freely, i.e., without any concern of reaching satu- The dissolved drug is continuously removed and the
ration in the medium. In the case of low solubility exiting solution is measured for the dissolved drug. If
drugs, solubilizing agents, such as sodium lauryl sul-
fate, are added to enhance the solubility. A low con-
centration of alcohol may be used to facilitate
dissolution for low solubility drugs. However, larger
concentrations of alcohol are to be avoided, as this
might result in misleading or unrepresentative results.
The dissolution tests are conducted in media kept at
a temperature of 37 C.

Stirring and mixing mechanism

The second major determinant for drug dissolution

testing concerns the device for stirring or mixing of Fig. 4 Schematic of USP drug dissolution apparatuses 1
the product with the dissolution medium. The instru- (basket) and 2 (paddle). (From Qureshi, S.A. Calibration:
mentation for mixing, in effect the dissolution appara- the USP dissolution apparatus suitability test. Drug Inf. J.
tuses, provides this aspect of the variant. Four different 1996, 30, 1060, Fig. 2.)
Tablet Testing 3713

vertical moving cylinder containing the remaining

product is lifted and moved to the next set of vessels
with fresh medium and the process is repeated a num-
ber of times. This instrument provides the strongest
agitation. Aside from the advantage of the system that
drug product can be tested at different pH values, the
system appears to have significant disadvantages. It
provides limited volume; thus, solubility of drug can
become an issue. There could be fluid carryover when
the product in the vertical moving tube is transferred
from one medium containing tube to the next. In
addition, automation of such a system for unattended
monitoring is not currently available.
Among the four apparatuses described above, most

Tablet–Tablet
of the experimental work reported in literature is based
on basket and paddle apparatuses.
Fig. 5 Schematic of a flow-through drug dissolution testing
apparatus, also referred to as apparatus 4 in the USP.
Reporting Results and Tolerances

Generally, dissolution results are reported as cumu-

required, the exiting liquid may be returned to the lated percent drug release vs. time. Presently, most of
medium reservoir for recirculation, thus providing a the tolerances are based on a single time point, such
closed-circuit operation with a limited or finite volume. as not less than 85% dissolved or released in 30 min.
However, its best use is achieved because of its open- However, more reports are appearing with percent
ended operation with a large dissolution medium vol- drug release values at multiple time points, resulting in
ume. The system is gaining popularity for products a drug release pattern, commonly known as a ‘‘profile.’’
of low solubility drugs. Its main disadvantage com- Commonly, dissolution tests are conducted in a set
pared to the paddle and basket apparatuses is that it of six units (tablets). Most commercially available
is somewhat more challenging to operate. apparatuses can run a test in a set of six units, thus
resulting in saving of time and resources and leading
Reciprocating Cylinder Apparatus. This system may to better reproducibility. Although tests are conducted
be envisioned as a collection of tubes/vessels in series, in a set of six, generally results are reported for individ-
as a set of six tubes. Each set of vessels contains a ual tablets. The most common form of tolerances is of
medium of similar or different characteristics such as the pharmacopeial ones such as the USP, which has
pH. A hollow cylinder (Fig. 6) with a wire gauze base been adopted by other pharmacopeias as well. These
contains the product and moves in a vertical motion in compendial standards are described in levels or stages,
each vessel. The dissolved drug is mixed with the exter- where each stage has its own set of tolerances that may
nal medium. At the end of preset time intervals, the be based on individual and mean results. For profiles, a
similar approach of tolerances is followed while con-
sidering every sampling time point as a discrete set of
results. It is important to note that there are no
requirements and standards of variance around the
mean such as standard deviation or coefficient of vari-
ation. A lack of such standards for variation appears
to be one challenge (weakness) in current dissolution
testing practice.[13,14]
Recently, a relatively new approach, known as
pooled sampling, has also been adopted for reporting
the results.[15] This approach is based on reporting a
single result obtained by pooling multiple samples,
usually six. The advantages of this approach appear
rather arguable. However, one significant disadvantage
is that it would hide variability in results from tablet to
Fig. 6 Schematic of a USP 3 dissolution-testing apparatus, tablet from a single lot or multiple lots, thus decreasing
also referred to as reciprocating cylinder apparatus. the quality of information obtained.
 3714 Tablet Testing

With regard to profiles, another parameter that Based on the drug solubility and permeability char-
has been introduced to compare drug release is (test acteristics, drugs may be divided into four groups: 1)
vs. reference product) known as similarity factor or high solubility–high permeability drugs; 2) low solu-
f2 factor.[16] In essence, it reflects a negative logarith- bility–high permeability drugs; 3) high solubility–low
mic sum of differences of percent drug release of two permeability drugs; and 4) low solubility–low per-
products over multiple sampling times. A value of this meability drugs. This classification of drug into four
parameter in the range of 50–100 is considered to groups is known as Biopharmaceutic Classifications
reflect similarity of profiles and thus similarity of pro- System.[25] Based on this classification system, one
ducts for drug release characteristics. However, there is would anticipate a successful IVIVC with drugs in
limited information currently available in support of low solubility–high permeability group. In this type
the usefulness of this approach. of drugs, as permeability is high thus as soon as drug
is dissolved, one would anticipate corresponding
absorption through the GI tissue. For the two cate-
Choice between In Vivo and gories that have drugs with low permeability values,
Tablet–Tablet

In Vitro Studies because the drug absorption is permeability dependent,

the IVIVC is unlikely to be observed. Furthermore, it
Both of these types of studies are conducted to assess is also unlikely that one would see IVIVC for drugs
the drug release characteristics of tablet products. in the category 1, i.e., high solubility and high per-
The gold standard remains in vivo studies. However, meability category. The reason being that if a drug is
as stated earlier, it is not necessary, due to ethical fast dissolving, e.g., in less than 15 min, then one would
and economical reasons, that one has to do the in vivo anticipate saturation or overloading of the GI tract
studies all the time. There are situations wherein the absorption system. Thus, changes in vivo dissolution
safety and efficacy aspect can be addressed by conduct- would unlikely be reflected in plasma drug concentra-
ing only the in vitro studies.[17,18] The confidence in the tions. However, if a high solubility–high permeability
in vitro results as a reflection of in vivo results is drug is formulated as a slow-release product, then
obtained if an established in vivo–in vitro correlation the drug would appear as low solubility/dissolution
(IVIVC) is available, i.e., in vivo drug release obtained drug and would behave as low-solubility–high per-
from plasma drug concentration–time profiles reflects meability drug where IVIVC should be possible.
the observed drug dissolution characteristics. There Therefore, IVIVC is not only a drug-dependent charac-
are numerous ways of observing and reporting teristic; it is also a product-dependent characteristic.
IVIVC.[19–23] However, for the purpose of reliability, Another reason for the lack of success of IVIVC
one generally compares the observed in vitro cumulat- may be due to the lack of relevant testing environment
ive drug release results with the cumulative drug in vitro. The currently used methodologies not only
release in vivo derived from the pharmacokinetic data. offer very poor reproducibility of results, but also very
Although there is significant enthusiasm in developing poor mixing and stirring characteristics in vitro.[14]
and reporting IVIVC in the scientific community, Therefore, it is highly unlikely that general success of
examples of successful IVIVC are rare for predicting IVIVC could be achieved soon. There is a clear need
in vivo characteristics based on in vitro results and vice for some improvement in this area.
versa. On the other hand, examples of lack of IVIVC Notwithstanding the above discussion, recently,
may be found more frequently. For example, in a there has been recognition of accepting in vitro results
study[24] to establish IVIVC, drug release characteris- in lieu of in vivo results for the assessment of safety
tics of various primidone tablet products were assessed and efficacy of pharmaceutical products. Particularly,
both in vitro (dissolution tests) and in vivo (BA/BE there has been a guidance document[26] from U.S.
study). The results of the evaluation indicate that even FDA that allows in vitro studies in lieu of in vivo stu-
though there were large differences in the in vitro dis- dies. In this particular example, if a drug has high per-
solution of the primidone tablets, the in vivo results meability and high solubility characteristics and no
were shown to be very similar. history of problem regarding the drug absorption,
In addition, one should also not expect an IVIVC inference to safety and efficacy of the product after
for all drug products. The drug absorption into sys- changes in product manufacturing can be granted pro-
temic circulation depends upon two aspects: 1) drug vided if the products show equivalent in vitro drug
release in the GI tract and 2) permeability characteris- release characteristics. The acceptance of in vitro
tics of the drug through GI tissue. If the absorption of results for in vivo is not based on IVIVC for this parti-
a drug is permeability dependent, then one should not cular group of products, i.e., products with high solu-
anticipate IVIVC because the IVIVC aspect only bility and permeability drugs and very fast in vitro
relates in vitro release to in vivo absorption or drug release characteristics (more than 85% dissolved in
appearance in the body. 15 min), but because experience with such drugs is that
Tablet Testing 3715

Table 1 An example of commonly used protocols for dissolution tests are relied on for the drug release
stability testing of tablet products characteristic in vivo. The analytical methods employed
Duration for stability testing must be validated to show that
Type Conditions (mo) the accuracy and precision have not been effected
Long-term testing 25
2 C/60% RH
5% 12 by the interferences of any potential degradation
product(s).
Accelerated testing 40
2 C/75% RH
6% 6

The stability studies on the active substances and
Alternate testing 30
2 C/60% RH
5% packaged dosage forms are conducted by ‘‘real time.’’
Alternate testing is required if significant changes occur during 6 mo Long-term tests at specific temperatures and humidity
storage under conditions of accelerated testing.
representing storage conditions experienced in the dis-
tribution chain of the climatic zone(s) of the country or
these have no BA problems and are unlikely to cause a region of the world concerned.
potential health risk. There are generally two types of stability studies
In short, it can be said that BA/BE testing remains conducted: 1) regular and 2) accelerated. In the case

Tablet–Tablet
the gold standard for establishing the pharmaceutical of accelerated stability study, the product is stored
ability of products in vivo. Due to ethical and eco- under elevated temperature and relative humidity
nomical reasons, BA/BE availability studies are mostly (RH) conditions to force or expedite its degradation
conducted at product development and major formu- pathway and assess the changes. The stability testing
lation and manufacturing process changes. However, conditions depend on the anticipated market environ-
an in vitro test, in conjunction with other analytical ment. An example of a recommended protocol[27] is
tests, is performed whether its relevance to in vivo is provided in Table 1.
established or not that appears to provide adequate
assurance of quality of products in particular antici-
pated release characteristics in vivo. Thus, drug disso- REFERENCES
lution testing has become an important and essential
quality control test for establishing lot-to-lot consist- 1. USP XXIV; The United States Pharmacopeial Convention,
Inc.: Rockville, MD, 2000; 1773–1774.
ency in the evaluation of products. Furthermore, in 2. British Pharmacopeia (BP) 2001; British Pharmacopeia
some cases such as for highly soluble and permeable Commission: London, UK, 2001; 1810.
drugs in fast release product characteristics, a dissol- 3. European Pharmacopeia, 4th Ed.; Council of Europe:
Strasbourg, France, 2002; 561.
ution test can also be used for a substitute of a BA 4. Specifications: Test Procedures and Acceptance Criteria for
study. New Drug Substances and New Drug Products: Chemical
Substances. International Conference on Harmonization
(ICH), Geneva, Switzerland, October 1999.
5. Gennaro, A.R., (Editor-in-Chief). Remington: The Science
and Practice of Pharmacy; Lippincott Williams & Wilkins:
PRODUCT STABILITY EVALUATION PA/MD, 2000; 883.
6. Gibaldi, M. Biopharmaceutic and Clinical Pharmacoki-
netics, 4th Ed.; Lea/Febiger: Philadelphia/London, 1991.
Product stability testing is an essential component of 7. Abdou, H.M. Dissolution, Bioavailability and Bioequiva-
establishing the quality of the products. The stability lence; Mack Publishing Company: Easton, PA, 1989.
parameters of a drug dosage form can be influenced 8. United States Food and Drug Administration Guidance for
Industry: Bioavailability and Bioequivalence Studies for
by environmental conditions of storage (e.g., tempera- Orally Administered Drug Products—General Consider-
ture, light, air, and humidity) as well as by the packag- ation, October 2000.
ing styles. 9. United States Food and Drug Administration Guidance for
Industry: Bioanalytical Method Validation, May 2001.
Knowledge of the physical stability of a formulation 10. Dressman, J.B.; Reppas, C. In vitro–in vivo correlations for
is very important for two reasons. First, a pharmaceu- lipophilic, poorly water-soluble drugs. Eur. J. Pharm. Sci.
tical product must look fresh and elegant as long as it 2000, 11, S-73–S-80.
11. Nichlaides, E.; Galia, E.; Efthymiopoulos, C.; Dressman,
remains on the shelf. Second, the active ingredient must J.B.; Reppas, C. Forecasting the in vivo performance of
be available to the patient throughout the expected four low solubility drugs from their in vitro dissolution
shelf life of the product. The causes of deterioration data. Pharm. Res. 1999, 16, 1876–1882.
12. USP XXIV; The United States Pharmacopeial Convention,
in quality of active ingredient could be due to Inc.: Rockville, MD, 2000; 1941–1951.
incompatibility with excipients, oxidation, reduction, 13. Qureshi, S.A.; McGilveray, I.J. Typical variability in drug
hydrolysis, and racemization to name a few. dissolution testing: study with USP and FDA calibrator
tablets and a marketed drug (glibenclamide) product. Eur.
With the exception of BA studies that are not J. Pharm. Sci. 1999, 7, 249–258.
conducted for the stability studies, all the above- 14. Qureshi, S.A.; Shabnam, J. Cause of high variability in drug
mentioned tests are performed after storing the pro- dissolution testing and its impact on setting tolerance. Eur.
J. Pharm. Sci. 2001, 12, 271–276.
duct in the package material to declare that products 15. USP XXIV; The United States Pharmacopoeial Convention,
of acceptable quality after the manufacturing. In vitro Inc.: Rockville, MD, 20001943.
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16. Shah, V.P.; Tsong, Y.; Sathe, P.; Liu, J.-p In vitro dissol- 22. Eddington, N.D.; Marroum, P.; Uppoor, R.; Hussain, A.;
ution profile comparison, statistics and analysis of the simi- Augsburger, L. Development and internal validation of
larity factor, f2. Pharm. Res. 1998, 15, 889–896. an in vitro–in vivo correlation for a hydrophilic metoprolol
17. Augsburger, L.; Shangraw, R.; Lesko, L.J.; Williams, R. An tartrate extended release tablet formulation. Pharm. Res.
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interpreting regulations and workshop reports of scale-up 23. Langenbucher, F. IVIVC: indices for comparing release and
and postapproval changes. Pharm. Res. 1994, 11, S-143. response profiles. Drug. Dev. Ind. Pharm. 1999, 25, 1223–1225.
18. Eddington, N.D.; Ashraf, M.; Augsburger, L.L.; Lesli, J.L.; 24. Meyer, M.C.; Straughn, A.B.; Mhatre, R.M.; Shah, V.P.;
Fossler, M.J.; Lesko, L.J.; Shah, V.P.; Rekhi, G.S. Identifi- Williams, R.L.; Lesko, L.J. Lack of in vitro/in vivo correla-
cation of formulation and manufacturing variables that tions of 50 mg and 250 mg primidone tablets. Pharm. Res.
influence in vitro dissolution and in vivo bioavailability of 1998, 15, 1085–1089.
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1998, 3, 535–547. theocratical basis for the biopharmaceutic drug classi-
19. Gillespie, W.R. Convolution-based approaches for in vivo– fication: the correlation of in vitro drug product dissolution
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Technology Transfer Considerations for Pharmaceuticals
Ira R. Berry
International Regulatory Business Consultants, LLC,
Maplewood, New Jersey, U.S.A.

INTRODUCTION program to one targeted toward commercialization.

Generally, the cost of product development rises
Technology transfer for pharmaceutical products is a dramatically during the pilot scale-up and initial pro-
program that has been followed for some time.[1–8] duction batch efforts. In other words, the critical
Issues to be considered when organizing the transfer of path for success is dependent on the completion of
technology from the research arena to the production the technology transfer to the production site at an
and quality assurance environments are reviewed in this affordable cost.
article. The discussion focuses on the coordination The three primary considerations to be addressed
and implementation of a transfer program for a product, during an effective technology transfer are the plan,
with emphasis given to those factors special to the the persons involved, and the process. A plan must

Technology–
pharmaceutical industry. The success of any program be devised to organize the personnel and the process

Tooling
is highly dependent on the effectiveness of the communi- steps. Once prepared, the plan must be communicated
cation preceding its implementation; therefore, the to the involved parties in research, at the corporate level,
preparation and distribution of a complete document and at the production site. The success of any program is
summarizing raw material and equipment requirements, highly dependent on the effectiveness of the communi-
manufacturing and packaging processes, process vali- cation preceding its implementation. Therefore, identi-
dation parameters, quality control procedures, and safe fying the parties involved in the development process is
handling procedures—as well as a detailed plan of action one of the most important tasks to be confronted and
outlining expected results and time frames—must be must be completed early in the transfer process.
distributed before the scale-up experience. Input from
the marketing and manufacturing centers must be
integrated into the plan to ensure that the right product
is developed at the right price within the desired time PERSONNEL
frame. An outline encompassing these critical aspects
of a transfer program is presented. The proper personnel must be informed of their
Whether a tablet, a transdermal patch, a topical involvement, desired contributions, and responsibili-
ointment, or an injectable, the transformation of a ties. This helps identify potential problem areas that
pharmaceutical prototype into a successful product may hinder the accomplishment of the challenge at
requires the cooperation of many individuals. To com- hand. It is desirable to appoint a project leader or liai-
plete the task efficiently, the transfer of a product from son from the research and development group as the
the research and development area to production must focus of the communication pattern. This individual
be organized. Planning for process commercialization has the responsibility to coordinate the assembly of
is one area where tangible rewards can be realized. the necessary information to support the product’s
The successful transfer of a project to a production site advancement for process development. The trap to
from the research arena does not happen on its own. avoid here is to assign someone to this role and not
Organizing the transfer of a technology or new product give him or her the authority commensurate with the
from research to production may be one of the responsibilities with which he or she has been charged.
most perplexing problems that development scientists, The practice of pairing a seasoned project manager as
engineers, and marketers may encounter during their a mentor with a less-experienced future project coordi-
careers. This article provides some insight into the nator is recommended. In this manner, team players
issues that should be considered during the transfer can be nurtured. The communication and networking
program and offers a sequence of events toward skills of the successful manager can be shared across
completing the task. the organization. On the other hand, it is expected that
A major decision focuses on that point where the individuals who do not possess the necessary talents to
idea or process is advanced from a research-oriented be effective are identified before they encounter the
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-100000441
Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved. 3717
 3718 Technology Transfer Considerations for Pharmaceuticals

stressful world of project management alone and, more The corporate office commonly involves personnel
importantly, before they have a chance to contribute to from the production planning unit, engineering group,
the failure of a potentially successful product. and new product coordination section, as well as from
Through mentoring, the one-to-one contact offers marketing, because each of these divisions has a vested
the unique opportunity for the sharing of ideas, skills, interest in the success of the venture. It is important
and observations. If properly organized with two-way to identify the needs of those people at the corporate
evaluations, both participants should benefit. Not all level to minimize delays caused by the ‘‘we did not
individuals, however, are suited for the role of mentor. know that we had to do this now’’ club. Activities such
Care should be exercised in the selection and pairing of as product label preparation, copy for advertisements
mentors for the less experienced. Perhaps a human and promotional pieces, and package graphics may need
resource manager could assist in identifying those to be initiated during the transfer program as not to lose
people with the interpersonal skills required for this valuable lead times. The acceptance or approval process
teaching position. in some companies is very labor-intensive and therefore
Information from the product development area time-consuming. This is one area where a detailed
would be gathered from the formulator, analytical time and event plan has been shown to yield a significant
and microbiology testing groups, and the packaging impact on timely completion of a project.
development unit. Issues to be considered from these Keeping the corporate participants informed becomes
groups will be discussed below. A safety evaluation a pivotal task for the project manager in obtaining the
from the toxicology group and industrial hygienist final acceptance of the product by the corporation.
should be completed before a scale-up effort. In today’s Therefore, any time taken with those people involved at
regulatory climate, a concise and understandable sum- the corporate level to explain the steps involved in the
Technology–

mary must be provided regarding the risks associated transfer program is time well spent. Like any educational
Tooling

with and procedures for proper handling of all chemical process, consistency and repetition enforce learning. It is
substances, whether drug actives or excipients. Failure incumbent on management to foster cooperation among
to provide sufficient information to those not skilled individuals at the functional research and corporate
in the art to make the decision to initiate a handling, centers, because the loss of time is the worst enemy an
weighing, or processing operation may lead to an organization can face.
unfortunate employee injury. Although an employee There are a number of individuals at the manufac-
has a ‘‘right to know,’’ employers have an obligation turing site who must work as a team to ensure the
to provide the necessary warnings and training to timely and efficient completion of this transfer effort.
minimize placing an employee at risk. The legal ramifi- The plant manager, technical director, production
cations of improper training or notification are outside planning group, manufacturing area supervisor, and
the scope of this article. quality control and quality assurance units—as well
An opinion should be solicited from the patent as the plant engineering, packaging and transportation
department or a patent attorney. With the implemen- supervisors—must be informed as to their responsibili-
tation of the General Agreement on Tarrifs and Trade ties. Personnel training must also be considered if the
(GATT) accord, patents take on a new meaning. The technology is new to the site. Last but not least are
impact of this legislation is far reaching, not only in the contributions that the line mechanics and chemical
the United States, where major revisions in the patent operators can make to the program. They perform the
laws have been required and implemented, but also in necessary production functions daily. Their insight and
many foreign countries where patents may be essen- practical experience are an invaluable resource that
tially worthless or not enforceable. In general, patents should not be overlooked.
have an effective life of 20 years from the date of initial The success of the transfer is dependent on the abil-
filing. Additional periods of exclusivity may be allowed, ity of the project leader to motivate employees to work
on application and when certain criteria are met, that toward a mutually beneficial goal, namely, introducing
extend the effective term of a patent. In summary, care a new product that will improve consumer health and
must be exercised to ensure that proper legal protection create jobs and increased profit for the company.
of the novel concept has been secured in those areas
where desired.
The drug regulatory affairs unit must also be involved MOTIVATING PARTICIPANTS
with the product transfer to specify ‘‘how much’’ of
‘‘what’’ information is needed either to submit for a drug Implementation of a positive return on involvement is
approval or to introduce a product to the marketplace. one mechanism by which changes in responsibilities
The goal here is to collect the proper amount and kind and tasks can be implemented. Changes here include
of data necessary to support the prerequisite filings, the manufacture of new products or using new pro-
either internal or external to the company. cedures to produce existing products. By involving
Technology Transfer Considerations for Pharmaceuticals 3719

personnel in the planning of changes, in discussing the The calendar pack and monthly cost to the consumer
resources necessary to complete the tasks, and in creat- not only position the product toward enhanced
ing an environment in which innovation can strive, consumer acceptance but also toward better patient
the project manager should strive to realize the com- compliance, especially with expensive medications. In
pletion of the program quickly and efficiently with contrast, inhalation aerosol units, because of their pat-
reliable quality at an affordable cost. At the same tern of chronic use, frequently contain sufficient doses
time, those who have worked to bring the program to last several months. Suffice it to note, it behooves
to completion should share a feeling of teamwork the marketing unit to work with financial analysts
and a sense of accomplishment. Return on involve- to determine the optimal product configuration and
ment encompasses the philosophy that employees are cost profile that maximizes consumer acceptance and
a key element in the successful introduction of any convenience, unit turns, profits, and resources.
technology or product approval. Project managers Constraints always exist but occasionally are not
must keep this in mind because they frequently depend communicated accurately or promptly. Competition
on the cooperation of individuals outside their direct in the marketplace frequently causes introduction
control to accomplish their goals. deadlines and endproduct cost constraints that must be
considered during the initial phases of a development
program. On the other hand, lead times for materials
PRETRANSFER CONSIDERATIONS and personnel resources must be appreciated when com-
mitments are made to timelines. Without planning in
Several assumptions must be satisfied before the advance, the task may not be possible at all. Factors that
advancement of any product to plant scale-up trials. influence decisions may originate externally as well as

Technology–
First, the marketing division should have examined internally. Care must be exercised to address those issues

Tooling
the proposed product prototypes and agreed that the that affect the timely completion of a project.
product meets their needs. Second, the intended com- Physicochemical properties of raw materials and the
mercial package configurations should have been selec- finished dosage form should be characterized before
ted. Although it is not uncommon to package portions any scale-up effort. Having methodology available and
of the first scale-up batch in a variety of formats, it is validated to compare batches is essential. For example,
incumbent on the project leader to eliminate unneces- drug-release profiles and viscosities have the potential
sary packages to minimize the dilution of effort. Third, of being altered in scale-up by manufacturing procedures
any constraints, such as cost or time, must be identified and equipment. Care must be exercised to maintain
so that they may be given due consideration. the desired profiles and other product specifications.
There is an important question to be asked, i.e., The effect of batch size and process scale-up should be
does the product meet the needs of the consumer? monitored closely.
The development staff believes they have captured Formulation and/or development of advanced
marketing’s vision with their product offering; how- drug-delivery systems such as microencapsulated mole-
ever, the marketing and sales units must concur. The cules, transdermal patches, or liposomes are frequently
decision is generally in the sales unit, because those accomplished in the laboratory. However, large-scale
people are responsible for making the product avail- production of these dosage forms may be problematic
able to the consumer and, more important, they are because the same conditions of manufacture may not
responsible to see that the product meets the consumer be attainable or desirable in the plant setting. Consul-
demands, real or perceived. It is expected that any tation with process development personnel during the
pertinent focus groups or market research studies finalization of the prototype development phase is
would be completed before the scale-up effort. These one way of minimizing scale-up difficulties.
studies help confirm the product concept and its viability An area occasionally overlooked by the develop-
in the marketplace. ment staff is the necessity of securing confidentiality
The cost of a development program increases dra- agreements from vendors supplying technologies or
matically as the number of package configurations to services to a firm. All contractors should be required
be advanced to commercialization increases. Selecting to execute a confidentiality agreement that specifi-
the proper package sizes, closures, colors, and neck cally encompasses the technology and product being
finishes is compromised by the package composition developed. These documents should be prepared,
and availability, and, finally, the intended use and cost reviewed, and executed by the appropriate legal and
of the unit. For example, a smaller unit lasting 1 executive officers of both organizations. Especially
month, such as a calendar pack, makes marketing when the science or product is not well-defined and
sense for oral contraceptives for several reasons. patent protection has not been secured, such as with
Dispensing and unit sales are generally cyclic and more the development of novel, specialized drug delivery sys-
predictable, allowing for better profit projections. tems or new chemical entities, this task must be completed
 3720 Technology Transfer Considerations for Pharmaceuticals

expeditiously, and no work should be initiated until compiled to ensure that appropriate consideration
the agreements have been properly executed. has been given to relevant issues. This helps ensure also
that all parties are approaching the task from the same
perspective and priority. A manufacturing site must be
TRANSFER PROGRAM designated and the appropriate personnel notified as to
their involvement. The necessary information must be
Any development and technology transfer program collected and disseminated to the involved parties. At
should be reduced to a written document such as a minimum, copies of the proposed formula, manu-
that shown in Fig.1. An outline or checklist must be facturing and testing procedures, and safe handling

Completion dates

Activity Target Actual

1. Formulation selected
2. Site for plant trial established
3. Planning meeting scheduled
a. Review of development report
b. Manufacturing formulation/procedures
b. Handling and safety issues
c. Raw material specifications and suppliers
d. Packaging procedures
Technology–

e. Packaging component specifications and suppliers

Tooling

f. Testing procedures and validation

g. Process validation protocols
4. Date of plant trial established
a. Manufacturing
c. Packaging
d. Quality assurance testing
c. On-site review of experience
d. Shipment of samples to R&D for testing
5. Date of shipment delivery
a. Confirmation of results
b. Stability initiation
c. Product evaluation
Safety
Efficacy
Drug release
6. Postproduction review meeting
7. Assembly of final pilot plant documents
a. Manufacturing
b. Packaging
c. Handling and safety procedures
d. Quality control specifications and analytical methods
e. Stability data
f. Material safety data sheet
g. Shelf-life projection
h. Process validation summary
i. Bibliography
8. Review of validation report
a. Research and development
b. Manufacturing
c. Engineering
d. Quality assurance/Quality control
e. Regulatory affairs
f. Corporate
g. Marketing
9. Issuance of final product specifications
10. First commercial batch

Fig. 1 Project technology transfer checklist. (Modified from Ref.[1].)

Technology Transfer Considerations for Pharmaceuticals 3721

considerations should be distributed to allow sufficient It would be appropriate to review the claims and
time for review and comment. A planning review physical characteristics of the product while evaluating
session should be convened with representatives from a sample so that the participants of a planning meet-
the research, corporate, marketing, manufacturing, ing appreciate the appearance and attributes of the
quality assurance, regulatory affairs, and engineering product.
departments in attendance. Selection of the time and The safety evaluation completed by the toxicology
location of this meeting should be made to encourage group and/or industrial hygienist should be reviewed
maximum participation. with the participants because they, as managers, will
The meeting should be chaired by the project leader. most likely assume the responsibility to protect the safety
It is his or her responsibility to determine the relevant of employees who will work on this project. The employ-
issues to be discussed, to follow up that an agenda for ees’ right to know must be protected further by keeping
the meeting is distributed, and to establish that minutes them informed of the potential risks to which they may
of the meeting are taken. Concerns that arise during be exposed.
this meeting should be noted and addressed because Prerequisite safety information can be transmitted
the purpose of the meeting is to draw from the experi- by draft material safety data sheets, especially for
ence of the participants to identify potential problem new drug actives and new drug products. Preparation
areas in the program. The tone of this session should of the draft document may lead to the question: Are
be one of consensus and not one of autocratic rule. the process handling procedures for this product neces-
Motivation, communication, and cooperation must sary and appropriate? Those personnel involved in the
be stressed in the voice and actions of the project lea- manufacturing of drug products are cognizant of the
der. This is the first step toward accomplishing the concepts of inherent risk due to an agent’s toxicity,

Technology–
primary program objective, namely, the timely and and potential risk, due to exposure. Whether the new

Tooling
informed transfer of a new product from the research drug and/or product should be handled in an open
arena to the production site. The following subjects environment, contained area, or in isolation must be
should be discussed at the planning meeting: determined by those people who are responsible for
safety. In all cases, the procedures must be reviewed
 Formula—handling and safety considerations and approved by the individual responsible for the
 Raw materials involved manufacturing and testing sites.
 Manufacturing equipment Comments about the prestability and finished
 Manufacturing precautions product stability profiles should be presented as an
 Manufacturing procedures overview of the new product’s chemical stability. This
 Packaging will support the anticipated shelf life of the formula
 Process validation and its ability to withstand the ‘‘process shocks’’ nor-
 Specifications for raw materials, packaging compo- mally encountered during production scale-ups. The
nents, and in-process and finished product toxicity of the finished product should be discussed.
 Validated analytical methods This information ensures that a decision regarding
 Regulatory considerations the handling of the formula has been made based on
 Rework procedures generated data and experience. A draft or tentative
 Transportation material safety data sheet may be one route to dissemi-
nate this information.
Each aspect should be reviewed to ensure that criti- Constraints on specific ingredients or sources of
cal issues have been addressed. If an aspect is not rel- ingredients, cost of goods, or manufacturing equip-
evant, it should be stated that it is not applicable to ment should be reviewed. Constraints must be con-
the program. An issue perceived as unimportant in sidered when formula optimization is undertaken;
one department may be a monumental task in another however, optimizing formulas may be best addressed
department. With new drugs and drug-delivery sys- in the production environment because batch size
tems, this effort is critical to the success of a program. and manufacturing equipment sometimes have been
shown to render viable laboratory and small-scale for-
mulas virtually inoperative in the plant. The experience
FORMULA gained during the manufacture of laboratory and
scale-up batches is invaluable and should be shared
Understanding the formula, its derivation, and its con- with the participants, especially the production and
straints is one of the first prerequisites to any develop- quality assurance staffs, in written reports and follow-
ment program. The feasibility of the formula may be up meetings.
established by reviewing the ingredients of the compo- Many firms use a laboratory development report to
sition and explaining their function in the formula. record these activities. These data can be incorporated
 3722 Technology Transfer Considerations for Pharmaceuticals

into a full project-development history. For new drug of the equipment fail, installation of a replacement part
products, development histories are needed to fulfill of known specifications reduces and in many cases
regulatory directives. The detail necessary in any sum- eliminates the need for revalidation of every product
mary report depends greatly on the magnitude of the processed with that equipment. The equipment itself,
problems encountered during development and the however, must be shown to meet its previous operating
corporate structure in which one lives. For some multi- capabilities before being placed back into service.
national companies, the product-development report The availability, size, and surfaces or composition
serves as the basis of spreading interest in a new of the required equipment should be specifically ident-
product across global borders. ified so that the scale-up effort may be representative
of a production run. A preliminary compatibility screen
of contact surfaces should be completed before the
RAW MATERIALS selection of scale-up equipment. The location of the
equipment in reference to other requirements, such as
Sources of raw materials, especially those critical to a services or the packaging area, may be a factor in the
new product’s functionality, should be identified. selection of equipment. A cleaning-validation study
Availability and costs should be ascertained to aid in should be conducted to ensure that no residues of active
the planning process. Care should be taken to ensure ingredient or cleaning agent remain after cleaning and
that new drug actives and, when possible, excipients that the equipment is suitable for production use again.
are secured from vendors with a current acceptable Alternative equipment may be considered and used;
FDA compliance profile and a drug master file. however, experience will dictate its suitability.
Testing monographs including methods to ascertain
Technology–

a lot’s chemical and, if necessary, microbiological

Tooling

integrity should be provided to the selected manufac- Precautions

turing site in advance so that the methods may be
applied to the incoming supplies. Handling of mate- Any concerns regarding the handling of equipment or
rials including storage, disposal, and employee precau- product by employees should be addressed in the plan-
tions should be documented, especially for new or ning stage. This is especially important with regard to
potentially hazardous materials. Again, material safety environmental (particulate contamination or sterility)
data sheets (MSDS) for all materials used should be or atmospheric (oxygen, moisture, or light sensitivity)
available and disseminated to the plant personnel problems.
before any exposure.

Procedures
MANUFACTURING
Procedures should be clear and concise. Specific descrip-
Equipment tions should be used when possible. For example, ‘‘pass
the emulsion through a suitable colloid mill (Eppenbach
When new drugs and drug-delivery systems are mill) at a setting of 0.005 inch (0.12 mm)’’ is preferred
developed in the laboratory, the correlation of the to a description referencing a more general description
necessary production equipment may be very difficult of a piece of equipment. Process-validation testing is
indeed. For example, the shear needed to create the necessary using specific equipment, as described by
desired particle size of an emulsion with the help of design and operating principle.
laboratory equipment may pose serious problems in Procedures should be realistic, and any instructions
the selection of plant equipment necessary to repro- must be scale-oriented. Specific parameters may be
duce the attributes of the product. Recording the speed necessary for manufacturing areas. For example, cool-
of a laboratory mixer is not sufficient by itself for ing or heating times are typically equipment-dependent.
this task; definition of the operating principle and Cooling 1 kg in the laboratory in 15 min may take
equipment design is necessary to accomplish the task. 4 hours for a 40,000-kg batch in the plant. Similarly,
Any equipment used in drug manufacturing, includ- filtration of small batches in the laboratory may not
ing packaging, should have undergone an equipment provide the necessary information to predict filter life
evaluation, including installation, operational, and per- or flow rates needed for large-scale manufacturing.
formance qualification, following a written protocol. Based on the experience gained during the pilot
This is an important step in a process-validation pro- scale-up effort, a process flow chart should be con-
gram. Through this effort, the operating parameters structed. It helps identify steps and issues in need of
and capabilities of a given piece of equipment are process-validation review. In addition, the timing of
documented. Furthermore, should a major component activities toward the scheduling of manpower needs,
Technology Transfer Considerations for Pharmaceuticals 3723

such as for in-process testing, is generally more appar- Whether it is for the development of new package
ent when viewed in the context of the total process. formats, such as for intranasal drug administration or
Process-optimization parameters as identified transdermal patches, or for more traditional delivery
during the pilot scale-up effort should be monitored systems, such as cycle packs, solutions, or aerosols, the
during the production scale-up batch. In this manner, need to educate employees involved in the processing is
appropriate recommendations based on experience essential to the transfer program’s success. Identifying
may be integrated into the production of future batches. and controlling process variables are necessary while
Many optimization experiments may be efficiently experience is gained, and the process is optimized and
incorporated into the process-validation program. validated.

Packaging PROCESS VALIDATION

The description, specifications, and test methods for Each class of product has specific issues to be
any packaging configuration should be available to addressed for process validation. In general, variable
the plant before the production scale-up. Unit func- process steps such as mixing times and temperatures
tionality and fit should be included as a practical use should be validated. Many articles have been written
test in any specification. The plant equipment to be regarding the validation of processes affecting pharma-
used in packaging should be evaluated for feasibility, ceutical products.[9–11] Protocols to evaluate those
speed, and contact surface compatibility. Preliminary parameters that may affect a product’s integrity should
evaluations of surface compatibility, discussed pre- be agreed on by the R&D, regulatory, production, QA,

Technology–
viously, should suffice as an early indication of packaging and engineering staffs. During the preparation and

Tooling
equipment suitability. packaging of the pilot production batch, generation
The availability of or lead time to secure the neces- of data toward improving the efficiency of these pro-
sary packaging components that are representative of cesses, as well as minimizing batch-to-batch variations,
the commercially marketed package frequently places is very important because this information will serve as
stress on the project timeline. Package costs and pos- documentation to support the new product’s commer-
sible alternative packaging can be evaluated with bulk cial feasibility. Also, the establishment of cleaning
produced from this batch. Therefore, a course of procedures and documentation of cleaning validation
action to minimize project failure resulting from an can be accomplished at this time.
unsatisfactory package may be appropriate. The result- There is no magic number of batches required to
ant dilution of resources and increased project expense prove that a process is validated. Generally, the num-
must be weighed in accepting this course of action. ber of batches accepted is three. The technical com-
The number of various package sizes to be filled plexities of and product sensitivities to variable
from a batch may be critical. For example, for generic parameters dictate how extensive a validation program
drugs, entire batches must be filled for the batch to be is needed. Suffice it to note that the process must be
accepted by the FDA. Although the batch may be filled controllable and reproducible and yield a product that
into numerous package formats, care must be exercised meets the desired specifications.
to fill a sufficient number of each format to ensure
proper equipment set-up and that the filling speeds used
are representative of a full-scale production effort. QUALITY CONTROL AND
Procedures for packaging the batch including fill QUALITY ASSURANCE
tolerances and precautions such as aseptic handling
or nitrogen gassing should be reviewed to state the One of the purposes of the pilot production batch is to
requirements for acceptance in advance. As a part of introduce the new product in its entirety to the func-
this production scale-up effort, it may be desirable to tional areas of the site (manufacturing, packaging,
evaluate the product’s bulk stability in the storage tank and testing), that is, the release of raw materials and
or storage drums to establish limits on the length packaging components as well as in-process, bulk,
of time a batch may be held before final packaging. and finished product testing should be completed at
This is especially important if the bulk product is the site as if the pilot scale-up were a commercial batch.
to be manufactured at one site and transported to In-process testing, bulk release before packaging,
another site for packaging. Storage container compati- and finished product specifications are proposed. Speci-
bility deserves appropriate attention also. fication limits are proposed based on the experience
Finally, the personnel involved in packaging should gained with smaller laboratory and scale-up batches.
be instructed on any safety and handling issues that Any comments or concerns regarding the test methods
might affect them or compromise the product’s integrity. and specifications should be addressed at this time.
 3724 Technology Transfer Considerations for Pharmaceuticals

Reagents and equipment to complete the required test- COMPLETION

ing must be available at the plant. A contact at the
R&D analytical laboratory should be established to In review, the activities to be completed at the manu-
explain aberrant values or to answer questions. facturing site are:
Communication of requirements of time and man-
power to the quality control department is a critical  Release of raw materials and packaging compo-
issue that must not be overlooked. Prompt attention nents.
to analysis needs does help keep the pilot production  Manufacture and packaging of the trial batch.
batch process moving forward. If microbiological  Generation of data from in-process, bulk, and fin-
release of bulk product is needed before packaging, ished product samples.
the project timeline should reflect the time period  Process validation, including equipment qualifi-
(3–5 days) generally needed for this activity. cation and reviews of batch records, processing,
Sampling must be scheduled for release and stability cleaning validation, and on-site experiences.
testing using a statistically valid sampling program.  Shipment of finished product to the research facility
This is especially important for stability studies. In this for testing.
manner, the chosen samples are documented to be
representative of the entire batch. An exit interview with involved plant personnel
Batch documentation is an important factor. Prep- offers an opportunity for their comments to be heard.
aration of master batch records in accordance with Their efforts should be acknowledged and their input
plant standard operating procedures (SOPs) should seriously considered and incorporated into the manu-
be followed by an approval of the document by the facturing document. Any differences that cannot be
Technology–

sponsoring division, usually the formulator or process resolved at this time should be noted and studied
Tooling

development staff of the R&D unit. On completion of further. The art of listening and diplomacy must be
a batch, review of the batch records by the quality used inasmuch as this forum must be one of
assurance group ensures compliance to GMP and that cooperation and not one of confrontation.
all necessary deviations from and modifications to the At this meeting, a discussion about possible rework
manufacturing records are properly explained and procedures may be appropriate. The ability to recover
documented. materials, especially expensive drug actives, is desir-
able. Early identification of steps where rework may
be possible allows for procedures to be tested, verified,
THE MANUFACTURING TRIAL and put in place, should they be needed. For pharma-
SCHEDULING DATE ceuticals, rework procedures may be used only if
they are appropriately documented, validated, and
At the conclusion of the planning meeting, any actions approved by the responsible corporate and govern-
that must be undertaken before the scale-up should be ment bodies. Rework procedures are not an automatic
documented and a responsible party identified. At the means for handling out-of-specification product lots
discussion of the trial date, time constraints must be but rather for identifying where an effort may be
considered, along with the availability of raw materi- implemented successfully. Logistics and economics, as
als, packaging components, plant scheduling time, always, dictate whether a rework should be considered.
and personnel. Coordination of personnel and supplies
is the responsibility of the project leader. The ability to
lead and negotiate another individual’s priorities helps POSTPRODUCTION ACTIVITIES
bring the trials to completion on schedule. AND EVALUATION
Finally, arrangements for transporting raw mate-
rials and packaged finished products must be made to Once the plant experience has been concluded, con-
ensure that the scale-up effort is completed on schedule firmatory analyses on duplicate samples for in-process,
and that stability studies are initiated expeditiously. bulk release, and finished product previously tested at
Participants should leave the meeting under the the plant site should be completed at R&D. In this
impression that one person is in charge of the project, manner, the proposed methods are challenged to yield
that the program has been well thought out and similar results from different analysts in different loca-
documented, and that commitments will be honored. tions. Discrepancies in values generated at this point
Early issuance of meeting minutes will reinforce the must be investigated and resolved. Samples should be
importance of individual responsibilities and serve placed into the stability testing system according to
as notice to the participants and their superiors that the organization’s procedures.
their cooperation has been solicited, is needed, and is Finally, a report must be prepared and issued
expected. expeditiously summarizing the experience, reviewing
Technology Transfer Considerations for Pharmaceuticals 3725

each area’s involvement, and proposing, if necessary, development histories and testing ‘‘quirks’’ regularly
changes in the process or control methods. Timely appear in these documents and in no other place.
and factual communication of project progress to the Skilled project leaders maintain an ongoing listing as
other corporate areas not directly involved in the a means of tracking a project. A bibliography for a
scale-up, such as the marketing, finance, and purchas- new pharmaceutical product may be voluminous, and
ing units, draws their attention and commitment of a reference to its location may be useful. The transfer
resources. By fostering informed decision-making of the project is considered complete when the first
through directed written communication, the time commercial batch is produced under the supervision
required to plan or complete these activities to bring of the manufacturing site without problems.
the product to market is minimized, and resource
usage is optimized.
A formal postproduction trial review meeting should CONCLUSIONS
be held with representatives from the R&D, corporate,
marketing, quality assurance, regulatory affairs, and This article has reviewed issues to be considered when
production centers. Plans to commercialize the product organizing the transfer of pharmaceutical technology
or to submit documentation for government approval from the research arena to the production environ-
if this is the next step in the development scheme are ment. Critical areas affecting the manufacture, packag-
outlined, contingent on the successful completion of a ing, safety, and quality of pharmaceutical products are
defined stability program. Agreement as to the suit- discussed in relationship to their impact by the tech-
ability of all the factors involved in the preparation of nology transfer process. The necessity of a plan with
the product should be the result of this meeting, with input from the various organizational centers is

Technology–
a substantiating document in the form of meeting min- emphasized. The success of the program is highly

Tooling
utes or a signed ‘‘statement of concurrence’’ generated dependent on the communication and cooperation
and distributed. shared throughout the process.
A monograph of all pertinent sections to support
the product’s introduction should be assembled,
ARTICLES OF FURTHER INTEREST
reviewed, and disseminated to the appropriate parties.
This document should include:
Pilot Plant Design, p. 2875.
Pilot Plant Operation, p. 2886.
 Manufacturing formula
Project Management, p. 3015.
 Draft label copy
 Raw material tests and specifications
 Manufacturing procedure REFERENCES
 In-process tests and specifications
 Finished product test methods and specifications 1. Popp, K.F. Encyclopedia of Pharmaceutical Technology,
 Packaging component specifications and drawings 1st Ed.; Swarbrick, J., Boylan, J.C., Eds.; Marcel Dekker,
Inc.: New York, 1996; Vol. 14, 419–432.
 Packaging component test methods 2. Owen, V.M. Technology transfer in the diagnostic industry.
 Stability data on bulk product Med. Dev. Technol. (Lond) 1994, 5, 23–26.
 Stability data on packaged product 3. Popp, K.F. Drug Dev. Ind. Pharm. 1987, 13 (13),
2339–2362.
 Shelf-life projection of expiration dating 4. Popp, K.F. Organizing the transfer of pharmaceuticals
 Material safety data sheet for the product from research to production. In Specialized Drug Delivery
 Bibliography Systems—Manufacturing and Production Technology; Tyle,
P., Ed.; Marcel Dekker, Inc.: New York, 1990; 37–50.
5. Rao, A.V.; Rajan, J.V. Transfer of drug technology from
A projection of shelf life should be included to laboratory to industry. Eastern Pharmacist (India) 1985,
document the recommendation for the expiration date. 28, 63–65.
6. Testa, E.G.; Lepiti, S. Product technology transfer. Phar-
Although accelerated stability data are frequently used maz. Ind. (Germany) 1981, 43 (12), 1231–1234.
to support expiration dating for up to 24 months, 7. Upupa, N. Research, product development, and pilot plant
extension of dating beyond 24 months is based on scale up. Pharmacy Times (India) 1990, 22, 63–65.
8. Evanoff, B.J.; Hofmann, K.L., Jr. Validation of Active
real-time test results. Pharmaceutical Ingredients, 2nd Ed.; Berry, I.R., Harpaz,
A bibliography of all project reports and memos D., Eds.; Interpharm Press: Englewood, CO, 2001.
should be assembled, including the development and 9. Berry, I.R. Process validation: practical applications to
pharmaceutical products. Drug Dev. Ind. Pharm. 1988,
validation reports. Ideally, the project leader should 14 (2 & 3), 377–389.
maintain a file in chronological order of all communica- 10. Berry, I.R.; Nash, R.A. Pharmaceutical Process Validation,
tions and reports. Compiling the file into a bibliography 2nd Ed.; Marcel Dekker, Inc.: New York, 1993.
11. Berry, I.R., Harpaz, D., Eds.; Validation of Active Pharma-
is a tedious task but one that, should the need to answer ceutical Ingredients, 2nd Ed.; Interpharm Press: Englewood,
a question arise, will be well worth the effort. Product CO, 2001.
 Thermal Analysis of Drugs and Drug Products
Danièlle Giron
Analytical Research and Development, Sandoz Pharma,
Basel, Switzerland

INTRODUCTION Differential Scanning Calorimetry (DSC)

Thermal analysis techniques, in which a physical pro- When a material is heated or cooled, there is a change
perty is monitored as a function of temperature or time in its structure or composition. These transformations
while the analyte is heated or cooled under controlled are connected with a heat exchange. Differential scan-
conditions, are fundamental techniques for the charac- ning calorimetry (DSC) is used for measuring the heat
terization of drugs and drug products, not only while flow into and out of the sample, as well as for deter-
processing or aging conditions may be simulated but mining the temperature of the thermal phenomenon
while the methods gives access to thermodynamic data. during a controlled change of temperature. The first
Due to the different informations delivered, thermal method developed by Le Chatelier in 1887 was differ-
analysis methods are concurrent or complementary ential thermal analysis (DTA), where only the tem-
Technology–

to other analytical techniques, such as spectroscopy, perature induced in the sample was measured.
Tooling

chromatography, melting, loss on drying, assay, for The principle of DSC is as follows: two ovens are
identification, purity, and quantitation. They are basic linearily heated; one oven contains the sample in a
methods in the field of polymer analysis and in physi- pan, the other contains an empty pan as a reference
cal and chemical characterization of pure substances as pan. If no change occurs in the sample during heating,
well as for mixtures. They find good applications for the sample pan and the reference pan are at the same
preformulation, processing, and control of the drug temperature. If a change such as melting occurs in
product. The introduction of automation considerably the sample, energy is used by the sample and the tem-
increases the advantages of these methods. New horizons perature remains constant in the sample pan while the
are open with the availability of combined techniques temperature of the reference pan continues to increase.
and microthermal analysis. Therefore a difference of temperature occurs between
the sample pan and reference pan.
Manufacturers use two methods of measurements.
PRINCIPLES AND EXPERIMENTAL In the first method called ‘‘heat flux DSC,’’ the instru-
FACTORS ment measures this temperature difference (DTA).
Through calibration, this temperature difference is
Considering the number of physical parameters of a transformed into a heat flow, dq/dt. Therefore, there is a
substance that may be measured, the number of techni- thermal factor that may vary with temperature. In the
ques derived is very large. Details on most techniques second method, called ‘‘power compensation DSC,’’
are well described by Wendlandt.[1] For pharmaceuti- two individual heaters are used in order to monitor the
cal applications, the methods generally used are differ- individual heating rates of the two individual ovens.
ential scanning calorimetry (DSC), thermogravimetry A system controls the temperature difference between
(TG) (or thermogravimetric analysis: TGA), and, to sample and reference. If any temperature difference is
a lesser extent thermomechanical analysis (TMA). All detected, the individual heatings are corrected in such a
techniques are automated and have data acquisition. way that the temperature is kept the same in both pans.
Hyphenated techniques and modulated DSC are grow- That is, when an endothermic or exothermic process
ing techniques, ‘‘state of the art’’ for the 21st century. occurs, the instrument delivers the compensation energy
Excellent books or review articles dealing with the in order to maintain equal temperature in both pans.
principle, instrumentation, and applications of thermal In the first case temperature is primarily measured,
analysis methods for pharmaceuticals are given in and in the second case, energy is primarily measured.
Refs.[1–14]. As emphasized by Cheng et al.[13], tendency The differentiation of measuring principles is with
in the next two decades willbemore precise and mean- modern instrumentation not very significant under
ingful measurements in these techniques and new normal applications. Due to calibration and integrated
developments in obtaining the temperature dependence data handling, the instruments produce similar quali-
of a material’s structure and dynamics. ties of reported results.
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-100200043
3726 Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved.
Thermal Analysis of Drugs and Drug Products 3727

Each instrument can deliver the same information, sample size is in the milligram range or less. Therefore
that is, heat flow as a function of temperature (or time). this correction is not very high, but it has to be taken
The peak shape, the resolution, and the sensitivity into consideration. Generally pure indium (>99.9999%)
depend on the principle of measurement and the speci- is used for the correction of the thermal lag.
fication of the instrument. Great efforts have been made in recent years in
For first-order transitions such as melting, crystalli- order to validate the different instruments not only in
zation, sublimation, boiling, etc., the integration of the comparing principle and results but also in determin-
curve gives the energy involved in the transition. For ing the critical parameters such as heating and cooling
second-order transitions, the signal gives the change rates, particle size, weight, resolution, atmosphere, and
in the specific heat, for example, glass transitions. type of pans (crimped pan, sealed pan, open pan, etc.).
Fig. 1 shows typical transitions. Melting and crystal- The instruments include automation and data
lization are first-order transitions. The extrapolated acquisition. The calibration of the instrument should
onset temperature (Te) is the melting or boiling point. be done at a yearly basis. This includes the measure-
The peak temperature (Tm) is dependent on instrument ment of temperature and enthalpy. Most certified
and measurement parameters. The glass point is standards are highly purified metals. Indium is the
determined as inflexion point. Manufacturers represent preferred reference standard, but it covers only one
the heat flow in different ways: the endotherms in the temperature. It is recommended for pharmaceuticals
positive side for power compensation DSC and in the to include several organic substances for which the
negative side for heat flux DSC. Melting, boiling, and melting point or the melting enthalpy has been accu-
sublimation are endothermic, which means they need rately determined. Sarge et al.[15,16] proposed several
energy. Crystallization is exothermic, which means that organic substances and metals. The heat determination

Technology–
it supplies energy. Desolvatations without melting are of quartz was also recommended. Sabbah et al.[17]

Tooling
generally endothermic. Solid–solid phase transition published recently a broad review of data of organic
and decomposition may be endothermic or exothermic. substances. For pharmaceuticals, it is suitable to have
Modern instruments provide heating, cooling and certified materials covering a broad range correspond-
isotherms between subambient temperatures (with a ing to the thermal events of interest.[18]
cooling device) and higher temperatures in the range Tables 1 and 2 are examples of calibration of the
of 1200–1500
C. In order to avoid reactions with the temperature and of the calorimetric response of PE-
atmosphere the measurements are carried out under DSC-7 instruments by using different materials.[19]
nitrogen. The major components of the systems Very important for pharmaceutical industry is the con-
include the DSC sensors, the furnace, the programer, fidence of the laboratory that delivers the reference.
and the data handling. The temperature plotted on Since the heating rate may have an influence on the
the abscissa is the programed temperature, not the data, it is recommended to compare the melting point
temperature of the sample. The difference between and the melting enthalpy of organic standards,
the programed and the actual temperature of the sam- additionally to indium, at different heating rates cover-
ple is called ‘‘thermal lag.’’ It depends on the thermal ing the measurement range. For very accurate determi-
resistance of the instrument and the heating rate. In nations, it is recommended to use standards with a
modern instruments dedicated to accurate analytical melting point in the range of the considered tempera-
measurements for pharmaceuticals, the sensors are in ture in a series of measurements.
direct contact with the bottom of the pans and the
Pressure DSC

In Pressure DSC (PDSC), the sample can be submitted

DSC Tm to different pressures, which allows to characterize
dq substances at the pressures of processes or to dis-
df
Endo

tinguish overlapping peaks observed, for example, by

desolvatation.[20]
∆CP Te
Isotherm
Exo

Modulated DSC
Crystallisation
Beginning of heating

Melting
Glass transition

This new technique introduced in 1993[21] has been

Oxidation
exotherm

thoroughly examined and discussed.[22] Main advan-

tages are the separation of overlapping events in the
DSC scans. In conventional DSC, a constant linear
heating or cooling rate is applied. In modulated DSC
Fig. 1 Theoretical DSC scans. (MDSC), the normally linear heating ramp is overlaid
 3728 Thermal Analysis of Drugs and Drug Products

Table 1 Example of calibration of Perkin-Elmer DSC-7 instruments with melting standards at 10 K min1 under nitrogen
Onset T (
C) Instrument 1 Instrument 2
Certified substances certificate onset T (
C) DT (
C) with intracooler onset T (
C) DT(
C)
Iodobenzene 31.3 32.2 0.9
H2O 0.0 0.1 0.1
4-Nitrotoluene 51.5 50.4 1.1 51.2 0.3
Biphenyl 69.3 68.2 1.1 68.6 0.7
Naphthalene 80.2 79.4 0.8 80.1 0.1
Benzil 94.7 94.21 0.6 94.5 0.2
Acetanilide 114.0 113.9 0.1 113.6 0.4
Benzoic acid 122.1 122.0 0.1 121.8 0.4
Diphenylacetic acid 146.5 146.9 0.4 146.9 0.4
Indium 156.6 156.8 0.2 156.5 0.1
Anisic acid 183.1 183.6 0.5 183.2 0.1
2-Chloro-anthraquinone 210.0 210.1 0.1 210.1 0.1
Tin 231.9 232.7 0.8 232.7 0.8
Anthraquinone 284.5 285.2 0.7 284.8 0.7
Lead 327.5 328.6 1.1 — —
Technology–

Zinc 418.9 420.3 1.4 — —

Tooling

with a sinusoidal function (MDSC) defined by a and (b þ Bo cos(ot)) is the measured quantity dT/dt
frequency and an amplitude to produce a sinusoidal- or ‘‘reversing’’ curve.
shaped temperature vs. time function. Using Fourier The total DSC curve, the reversing curve giving
mathematics, the DSC signal is split into two reversible transitions and the non-reversing curve giv-
components: one reflecting non-reversible events ing irreversible transitions (e.g., the glass transitions),
(kinetic) and the other reversible events. is obtained. MDSC is a valuable extension of con-
ventional DSC. Its applicability[23] is recognized for
T ¼ T0 þ bt þ B sinðotÞ precise determination of the temperature of glass tran-
dq=dt ¼ Cðb þ Bo cosðotÞÞ þ f 0 ðt; TÞ þ K sinðotÞ sitions and for the study of the energy of relaxation,
and it depends on a number of important parameters
where T is temperature, C the specific heat, t the time, to be studied. It has been recently applied for the
o the frequency, f 0 (t, T) is the average underlying determination of glass transitions of hydroxypropyl-
kinetic function once the effect of the sine-wave methylcellulose films[24] and for the study of amor-
modulation has been substracted. K is the amplitude phous lactose,[25] as well as for the study of some
of the kinetic response to the sine-wave modulation glassy drugs.[26]

Table 2 Examples of calorimetric measurements of standards with two different DSC-7 instruments and measurement cell
at different times (A, B, C) at 10 K min1 under nitrogen
A B C

Standard substance DH (J/g) (Theory) DH (J/g) % deviation DH (J/g) % deviation DH (J/g) % deviation


Naphthalene (80.2 C) 148.6 147.1 1.0 148.6 0.0 — —
Benzil (94.7
C) 112.0 110.1 1.7 112.8 0.7 — —


Benzoic acid (80.2 C) 147.2 — — — — 146.6 0.4
Biphenyl (69.3
C) 120.4 120.0 0.6 — — 120.5 0.1
Diphenyl-acetic acid (146.5
C) 146.9 — — 146.8 0.1 — —
Indium (156.6
C) 28.7 28.63 0.2 28.8 0.35 28.7 0.1
Tin (231.9
C) 60.2 60.0 0.3 — — 60.8 1.0
Thermal Analysis of Drugs and Drug Products 3729

Microwave thermal analysis (MWTA) 5.0

1st Derivative (%/min x 10–1)

100.0
0.0
95.0
In this new technique,[27] microwaves are used both to ∆Y–15.74 Wt. %

Weight (Wt. %)
–5.0
90.0
heat a material and to detect thermal transitions. 85.0 ∆Y–28.55 Wt. % –10.0
∆Y–35.99 Wt. % –15.0
80.0
Micro-DSC 75.0 –20.0
70.0 –25.0
The instruments of conventional DSC allows to mea- 65.0 –30.0
sure very small amounts of material. The author was 60.0
able to characterize the melting peak of indium with 50.0 100.0 150.0 200.0 250.0 300.0
0.032 mg by using a DSC-7 of Perkin-Elmer. New Temperature (˚C)
instrument generation will permit to increase sensi-
Fig. 2 TG of copper(II) sulfate pentahydrate with the heat-
tivity and amount of material to be studied decrease
ing rate 20 K min1. Use of DTG for the different steps.
to nanorange.[28]

Microcalorimetry
decomposition follows desolvatation by use of the
[29,30] minima in the DTG curve.
Microcalorimetry is a growing technique com-
The instrument used in thermogravimetry is a
plementary to DSC for the characterization of
thermobalance (balance controller, sample chamber,
pharmaceuticals. Larger sample volume and high sen-
furnace, furnace controller) with data processing. In
sitivity means that phenomena of very low energy

Technology–
order to check the stability of the system a baseline

Tooling
(unmeasurable by DSC) may be studied. The output
at the highest sensitivity has to be done for all heating
of the instrument is measured by the rate of heat
rates in the temperature range of analysis: The highest
change (dq/dt) as a function of time with a high sensi-
deviation will be observed at the highest heating rate.
tivity better than 0.1 mW. Microcalorimery can be
The thermobalance may have a vertical or horizontal
applied to isolated systems in specific atmospheres;
construction. The sensitivity of new thermobalances
or for batch mode where reactants are mixed in the
attains 0.1 mg. Some manufacturers offer combined
calorimeter. Solution calorimetry can be used in
DSC/TG instruments.
adiabatic or isoperibol modes in microcalorimeters at
The mass accuracy is generally not a problem of
constant temperature. (See the corresponding article
modern TG. Calibration of the mass with certified
about calorimetry of this edition.)
mass can be used as for all other balances. Electro-
statics, temperature fluctuation, sensitivity of the
Thermogravimetric Analysis sensor, and thermal lags have to be known, what is best
done with regular calibration. For automatic TG, the
In thermogravimetry (TG or TGA) the change in sam- pans have to be tightly closed and pierced just before
ple mass is determined as a function of temperature the measurement; therefore, the TG curves of desolva-
and/or time. The instrument is a thermobalance that tation may be different as for open pans. The use of a
permits the continuous weighing of a sample as a func- protective gas and its flow, as well as the sample
tion of time. The sample holder and a reference holder mass and the heating rate play a role in the comparison
are bounded to each side of a microbalance. The sam- of the temperature of thermal events. The influence
ple holder is in a furnace, without direct contact with of heating rate is examplified in Fig. 3 with
the sample, the temperature of which is controlled by CuSO4  5H2O. The limit of detection can be calculated
a temperature programer. The balance part is main- by determining the maximum of deviation of the base
tained at a constant temperature. The instrument is line in the temperature range of interest.
able to record the mass loss or gain of the sample as Table 3 shows an example of calibration performed
a function of temperature and time [m ¼ f(T)]. Most with hydrates, which cover the starting of dehydration
instruments also record the DTG curve, which is the temperature from 50 to 120
C and the end of dehy-
rate of the mass change dm/dt ¼ f(T). dration from approx. 150
C until to 270
C with differ-
The DTG curves, allow a better distinction of ent heating rates.
overlapping steps, as demonstrated in Fig. 2, for Since there is no contact between pan and furnace,
CuSO4  5H2O. The area under the DTG curve is pro- the thermal lag is higher than in DSC. The standards
portional to the mass change, and the height of the recommended by ICTA and distributed by NBS are
DTG peak at any temperature gives the rate of mass ferromagnetic standards exhibiting loss of ferro-
change. The real advantage of DTG is to permit accu- magnetism at their curie point temperature within a
rate location of the end of a desolvatation process if magnetic field: Nickel (354
C), Permanorm 3 (266
C),
 3730 Thermal Analysis of Drugs and Drug Products

101.4 Perkin Elmer Thermal Analysis Thermomechanical Analysis

100
95 In thermomechanical analysis (TMA) the deformation
90 of the sample under stress is monitored against time
Weight % (%)

85 or temperature while the temperature increases or

80 decreases proportionally to time. Changes are detected
75 by mechanical, optical, or electrical transducers. The
70 stress may be a compression, penetration, tension,
flexure, or torsion. Generally the instruments are also
65
60.58
able to measure the sample dimensions, a technique
22.41 50 100 150 200 250 300 called thermodilatometry. The stress (F/A) expressed
Temperature (˚C) in N/m2 or Pa may be a normal tensile stress s, a
tangential shearing stress t, or a pressure change Dp;
Fig. 3 TG of copper(II) sulfate pentahydrate. Influence the force applied is F and A is the area.
of the heating rate. TG curves from the top: 20 K min1, The deformation is measured by the strain, which is
10 K min1, 5 K min1, 2.5 K min1. the deformation per unit dimension.

Numetal (386
C), Permanorm 5 (459
C), Trafoperm

Elongation e ¼ DL=L0
(754
C). The method does not permit the temperature
measurement with high precision. These standards Volume strain y ¼ DV =V0
have been studied by several authors.[31] The ICTA Shear strain g ¼ Dx=y
Technology–

temperatures are within 5–10
C. McGhie et al.[32] pro-

Tooling

posed a calibration technique in which a small inert

platinum weight is suspended by a fusible link com- For an elastic material, the Young’s modulus is defined
posed of a calibration standard that releases the plati- by
num weight at the temperature of melting. The Mettler
instrument TGA 850 is constructed so that the melting
curve of standards can be measured and used as cali- E ¼ ðF=AÞ=ðDL=L0 Þ or E ¼ s=e
bration, as demonstrated in Table 4.
TG can be used with different atmospheres and Creep is the gradual irreversible elongation of the
under vacuum. TG has a huge number of pharmaceu- sample.
tical applications. Automated TG is extremely efficient These parameters depend on the temperature. The
to replace the loss on drying assay in drug substances, coefficient of thermal expansion is
being able to separate loss of solvent from decom-
position by using very small amounts of substance.
  
Solvent entrapped or bounded as solvate is easily dL l
determined.[9,33,34] A comprehensive article on TG a ¼
dT L0
has been recently written by Dunn and Sharp.[35]
Ozawa proposes the use of modulated TG for kinetic
analysis.[36] The instruments have a furnace, and mostly a linear
Water sorption–desorption isotherms can be carried variable differential transformer (LVDT) to produce
out by using thermobalances. Now specific instruments an electrical signal from a linear movement. An
allow to measure water sorption–desorption isotherms additional unit controls the force applied. Special
at different constant temperatures (e.g., dynamic vapor attachments allow the same instrument to work in
sorption instrument (DVS), Surface Measurement different modes such as elongation, compression,
Systems Ltd., Monarch Beach, US). penetration, or tension.

Table 3 Example of calibration of loss of mass with three hydrate standards

Result

Substance Theoretical amount water 5 K min1 10 K min1 20 K min1

Sodium-tartrate dihydrate 15.7% 15.73% 15.60% 15.73%
Calcium-oxalate monohydrate 12.3% 12.55% 12.51% 12.48%
Copper-sulfate pentahydrate 36.1% 36.08% 36.03% 36.04%
Thermal Analysis of Drugs and Drug Products 3731

Table 4 Example of calibration of the temperature of TGA 850 with melting standards
Result
1
Substance Theory 5 K min 10 K min1 20 K min1
Nitrotoluene 51.5
C 51.49
C 51.64
C 53.78
C
Indium 156.6
C 157.62
C 157.38
C 157.74
C




Tin 231.9 C 233.44 C 233.42 C 233.68
C

The slope of the TMA trace may also be obtained ratio of these modulus, is plotted against temperature.
by DTMA. A recent overview of the pharmaceutical applications
of DMA has been published by Craig and Johnson.[41]
dL dT Torsional braid analysis (TBA) is a particular case
¼ L0 a
dt dt where the sample supported by a fiberglass braid is
where dT/dt is the heating rate. subjected to a torsional strain.
Thermomechanical methods are very useful for the
determination of phase transformations such as poly- Hyphenated Techniques
morphic solid–solid transitions or glass transitions. (Combined Techniques)
Fig. 4 shows some theoretical curves for glass tran-
sition and polymorphic transition in extension or in A comprehensive characterization of the physical

Technology–
penetration mode. Recently TMA has been proposed properties of materials often requires a multidisciplin-

Tooling
for the measurement of the internal stress of tablets ary approach since no single technique is capable of
of ethylcellulose of different molecular weight[37] and characterizing pharmaceuticals completely.
for measurement of swelling of polysaccharide hydro- Thermomicroscopy or hot stage microscopy is a
gels[38] and of polymeric films.[39] well-established method[42a,42b–44] for the microscopic
Calibration of the instrument for its response to observation with polarization of the sample while heat-
length may be carried out with a standard length piece ing or cooling, allowing to see desolvatation, melting,
of metal or ceramic. The temperature can be calibrated crystallization, eutectic formation, and even transfor-
in the same way as for DSC. Metal standards such as mations in suspensions in solvents. The combination
indium, tin, or lead are mostly used. Recent publica- of hot stage microscopy to new technology such as
tions[40] deal with calibration and errors of TMA. high-resolution color camera, image manipulation
software makes the technique very attractive for
inducing metastable states, for observation of crystal
Dynamic Mechanical Analysis habit, and for better interpretation of other methods.
Thermophotometry is the measurement of the light
In this technique, the mechanical response of a sample intensity and thermoluminescence of the light emitted
is measured as it is deformed under oscillating load by the sample. FT-IR microscopy,[14,44,45] Raman
against temperature or time. Dynamic mechanical microscopy[46–48] are excellent additional tools to ther-
analysis (DMA) is a further development of TMA, momicroscopy. Calibration of microscope, of heating
but the instruments are different. unit, and of spectroscopic methods should be done.
DMA is mostly applied to the study of polymers. TEM (transmission microscopy) and SEM (scanning
Relevant parameters are the storage modulus and the electron microscopy) with EDX have been combined
loss modulus. Generally the loss tan d, which is the to DSC for the study of solid-state reactions.
Newly born, the scanning thermal microscopy
derived from atomic force microscopy brings a revolu-
Expansion Expansion L
tion in the instrumentation for measuring thermophysi-
Tg Melt cal and thermomechanical properties of the matter,
and the TA instrument was awarded at Pittsburg 1998.
Transition The instrument has been applied for the characterization
Tg
of Ibuprofen compacts as model substance.[49]
Temperature-resolved X-ray diffraction with a heating
T ˚c T ˚c T ˚c
cell is widely used.[11,50–53] Crystalline changes are clearly
Mode: penetration Mode: load Mode: load
assigned; the X-ray diffraction patterns obtained in situ
Fig. 4 Theoretical TMA curves for glass transition Tg and allow to predict quantitative methods if, for kinetic rea-
polymorphic transition. sons, forms that are present at high temperatures occur
 3732 Thermal Analysis of Drugs and Drug Products

at ambient conditions. Low-temperature X-ray diffrac- Thermogravimetry can be coupled with DSC.
tion cell has been developed for the study of frozen aque- Most companies offer the TG-IR[58] or the TG-MS
ous solutions.[54] The introduction of XRD-DTA cell[55] coupling.[59] Synergic chemical analysis by coupling
and recently of the DSC-XRD instrument of Rigaku TG-FT-IR, TG-MS or TG-GC-MS has been recently
presented at the Denver X-ray Conference in 1999[56] discussed.[60] Fig. 5 demonstrates the ability of TG-
demonstrates the advantage of this direct combined MS for the study of dehydration and decomposition
technique. The observation of polymorphic transforma- of calcium oxalate dihydrate. The steps correspond
tion by using variable temperature synchrotron X-ray to the dehydration into anhydrous calcium oxalate,
diffraction method is a promising technique with the followed by the transformation into calcium carbonate
new computerized ability for obtaining structural data.[57] then by the formation of cacium oxide.[10] The sample

100

90

80

70
Technology–
Tooling

60

50

40
100 200 300 400 500 600 700 800 ˚c

0 5 10 15 20 25 30 35 40 min
Oxalate

Ion Current [E-07A]

0.16000

0.15000
Calcium Oxalate lot K11H09
0.14000

0.13000

0.12000
Water
0.11000

0.10000

0.09000 CO

0.08000

0.07000

0.06000

0.05000

0.04000 CO2
0.03000

0.02000

0.01000

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42
l [min]

Fig. 5 TG and TG-MS of calcium oxalate monohydrate. The TG steps correspond to the dehydration followed by the forma-
tion of calcium carbonate, then calcium oxide is obtained. The evolved gas is detected by MS.
Thermal Analysis of Drugs and Drug Products 3733

studied in Fig. 5 contains additionally to crystal water example of polymorphism is carbon, which can exist
some free water. The TG-MS shows that some amount in the form of graphite or as a diamond.
of the CO evolved during the decomposition also The amorphous state characterizes crystallization in
transforms into CO2. a non-ordered, random system, related to the liquid
These new emerging combined techniques enables state. The name ‘‘glassy state’’ is given to amorphous
the observation of extremely small samples with a high products that liquify by undergoing a glass transition.
degree of information. They find a good place for The expression pseudopolymorphism applies to
proper screening[54] (according to the polymorphic stud- hydrates and solvates.
ies required by ICH)[61] in order to choose the proper Different solid phases that may occur during crys-
form and to justify its choice. They also permit to ana- tallization or galenical processes are polymorphs,
lyze more easily the complex matrixes of drug products. amorphous phases, or solvates as the result of com-
pound formation with the solvent.[62–65]
A recent detailed review about thermal analysis of
APPLICATIONS OF THERMAL ANALYSIS polymorphs and pseudopolymorphs[66] listed more
TECHNIQUES FOR DRUG SUBSTANCES than 300 drug substances presenting this behavior in
the literature. Polymorphism of excipients and their
The transitions observed by thermal analysis techni- thermal analysis has been reviewed in Ref.[67].
ques are based upon the Gibbs phase rule and phase DSC gives not only temperature of events, but also
diagrams, p, T, concentration. melting energies. The Burger’s rule and the energy dia-
All transitions or reactions involving energy grams[68–73] help considerably to approach the thermo-
changes may be measured by DSC. Transitions involv- dynamic equilibrium of a polymorphic system.

Technology–
ing mass changes are detected by TG. For a single For each polymorph (single compound), there is a

Tooling
product, specific heat, glass transition, melting, boiling, solid–liquid equilibrium curve and a solid–gas equilib-
sublimation, decomposition, and phase transitions rium curve. The solid–gas curves meet at a point. If the
induced by polymorphism during heating are impor- liquid–gas equilibrium curve meets the two solid–gas
tant for the choice of the salt form and for safety stud- curves after this point of intersection, there will be a
ies where the DSC exothermic peaks are relevant. The solid 1, solid 2 equilibrium curve and a reversible tran-
use of DSC for the measurement of the melting point sition point 1$2 at a specific pressure. This is known
of raw materials has been proposed.[19] Hydrates or as enantiotropy. At the transition point, the free energy
solvates, or volatile compounds in the formulations of the two forms is the same.
can be investigated by DSC combined with TG and The term monotropy applies in the case of an
TG-MS. DSC curves of mixtures of solid compounds irreversible transition from one form to another.
depend upon the phase diagrams in solid state. If Monotropy is bound to the existence of metastable
there is no interaction in the solid state and if there thermodynamic forms. The liquid–gas curve crosses
is a miscibility in the melt, an eutectic behavior is the solid–gas curves for the two forms before their
observed. This enables the purity determination of point of intersection.
raw materials, the analysis of enantiomers, and the Knowing the relationship between the thermo-
study of ‘‘physical interactions’’ in preformulation. If dynamic quantities H (enthalpy), G (free energy), S
the compounds are not miscible in the liquid state, (entropy), and T (temperature), it is often simple to
the DSC curve of the mixture is the addition of the represent equilibrium states by plotting the free energy
DSC curves of each compounds. Interaction is G as a function of the temperature for each form. If the
observed in the solid state in case of formation of solid two curves intersect before the melting point, there is
solution or complex formation between components or reversibility, i.e., enantiotropy, and if the reverse is
in the case of chemical reaction. true, there is monotropy.
The relationship between melting enthalpies of two
solid phases A and B and the heat of transition is:
Polymorphism/Pseudopolymorphism
and Amorphous State DHt ¼ DHAf  DHBf

Polymorphism is the tendency of any substance to Figs. 6A and B illustrate the plots of the functions
crystallize into different crystalline states. The solid G and H versus temperature (energy/temperature
forms of the same compound are called polymorphs diagrams) for each polymorph and for the liquid.
or crystalline modifications. On melting, they produce The thermodynamic reversibility of the solid transition
the same liquid. Polymorphs show the same properties between two crystalline forms is characteristic of
in the liquid or gaseous state but they behave as differ- enantiotropic systems. Each form has its thermo-
ent substances in the solid state. The best known dynamic stability range. The lower melting form is
 3734 Thermal Analysis of Drugs and Drug Products

A B
Energy (G, H) Energy (G, H)
HL
HL
∆Hf,B
∆Hf,A
HB ∆Hf,B
∆Hf,A HB
HA
∆Ht HA
B
B
A
GA A
GB
GB
GA
GL GL
Tt Tf.A Tf.B Tf.B Tf.A
Temperature Temperature

Fig. 6 Energy diagrams showing plots of enthalpy H and Gibbs free energy G, against temperature T, for the solid and liquid
phases of a single compound, showing (A) enantiotropy and (B) monotropy.

stable in the temperature range below the transition In the case of enantiotropy (Fig. 7A), only the form
Technology–

point; the higher melting form is stable in the tempera- A should be encountered below the transition point
Tooling

ture range above the transition point. In case of mon- and the behavior upon heating is illustrated by the
otropy only one form is stable whatever the DSC scan 1, the endothermic transition A () B
temperature range. The Burger’ rule is as follows in should be observed followed by the melting peak of
the case of enantiotropy, the lower melting form has the form B. For kinetic reasons, e.g., in case of too
the higher melting enthalpy and the transition into quick heating rate, the transition A () B does not
the high melting form by heating is endothermic; in occur and form A melts. Then two possibilities may
the case of monotropy, the thermodynamic stable form be found: no other signal occurs (DSC scan 2) or an
is the higher melting form with the higher melting exothermic crystallization of B from the melt occurs
enthalpy. The transformation of the unstable form into and later on, the melting peak of B is observed (DSC
the stable form is exothermic. scan 3). If the metastable form B occurs below the
Because of kinetic factors, metastable forms are transition, upon heating it can be transformed into A
encountered in temperature ranges outside the ther- with an exothermic transition (DSC scan 4); thereafter,
modynamic range. Crystallization processes generally the form A transforms into the form B according to
imply the cooling of concentrated solutions or precipi- scan 1 or scan 2). Finally (scan 5), the metastable form
tation by addition of cosolvent. Depending on the rela- B can melt without any transformation.
tive positions of the solubility curves of the metastable In the case of monotropy (Fig. 7B) only the form
polymorphic forms and the metastable curve of super- A should exist and the DSC scan should show the
saturation, the first nucleous can be a metastable form. melting peak of the stable form A (scan 1). If the meta-
Transformation to the stable crystalline form may or stable form B is heated, then scan 2 or 3 may be
may not occur, depending on kinetic factor. Furthermore observed: the form B transforms exothermically into
solvates exist at lower temperatures and their presence the stable form A (scan 2) or the form B melts and
should be considered and finally due to the humidity of the stable form A grows from the melt and its melting
the air or from water activity of the solvents, hydrates peak is observed.
may be formed. Polymorphism of solvates and hydrates In the case of racemate, the situation is more com-
is not uncommon. This phenomenon of concomittant plex since one racemate polymorph can be a true
polymorphs has been recently reviewed.[74] racemate and the other one a conglomerate[72,73] and
Fig. 7 illustrates the behavior of polymorphs A and one has to consider additional solid phase transitions
B in case of enantiotropy (Fig. 7A) and monotropy such as peritectoids and eutectoids, which are not
(Fig. 7B) during heating. For all analysis where a polymorphic transitions.[75]
temperature change is involved, kinetic factors have to Very often some substances have two melting
be considered for proper interpretation of the results. points separated by an exotherm. Such a DSC scan
The DSC scans will differ if the sample being analyzed can correspond to a monotropy or to an enantiotropy.
is stable or metastable at ambient temperature. A is the The sample may be a pure form or a mixture. Using
stable form at ambient temperature in both cases. different heating rates and tempering in DSC, one
Thermal Analysis of Drugs and Drug Products 3735

A The resulting degradation product then recrystallizes

and melts at higher temperatures. We observed such
an effect for aspartam[12] and for a malonate salt,
which decomposed in the corresponding base.[11] If
A B L
1 an isomerization takes place, then only the analytical
data of the product obtained allow us to have an accu-
rate interpretation.
These curves illustrate the complex transitions that
A L
2 may occur when heating or cooling polymorphs.
Therefore, combined techniques are very useful for
Heat flow dq/dt Endo

complete interpretation of observations given by

A L
DSC as emphazised in Ref.[53]. Fig. 8 illustrates a
3 reversible enantiotropic transition followed by DSC
and by temperature-resolved X-ray diffraction of a
B
purine derivative. For this substance six crystalline
B A B L forms were found. The Burger’s rule as well as the
4 study of the supersaturated solutions and the use of
combined techniques allowed us to find out that the
form of Fig. 8 was the stable form below the transition
B L point. All other forms were monotrops to this form.[77]
5 Table 5 deals with the example of a benzisoquino-

Technology–
line hydrochloride for which both forms presented a

Tooling
T T T
t f,A f,B melting that was followed by decomposition. No
Temperature change was observed by slow heating rate. Since the
B
melting enthalpies differed only by 10%, the proper
interpretation needed the verification of the hypo-
thesis: enantiotropic transition. The analysis of the
insoluble solid in the equilibration of both forms in
A
alcohols (solvent mediated transition) showed that
1 L
form A is always obtained, what confirmed the
Heat flow dq/dt Endo

observation of the Burger’s rule.

B A L
2 Amorphous State

If a physical property of a crystalline substance is

plotted against temperature, a sharp discontinuity
B A L
3 occurs at the melting point. For amorphous sub-
stances, there is no melting point, and a change of
T T
slope occurs at the so-called glass transition tempera-
f,B f,A ture Tg. The glass transition is characterized by a
Temperature change of heat capacity. Below this temperature, the
amorphous phase has certain properties of a crystalline
Fig. 7 Possible DSC curves for two polymorphs: (A) Enan-
solid (e.g., plastic deformation) and is termed ‘‘glassy.’’
tiotropy A (()) B, B is the highest melting form, A is the
stable form below the transition point. (B) Monotropy
B ! A, A is the highest melting form. (For explanations of Table 5 Study of two polymorphs with unique melting
the scans, see text.) curve
Crystalline Crystalline
may be able to measure melting points and melting Property form A form B
enthalpies and to use the Burger’ rule. Melting point
C 304 311
The TG curve is extremely valuable for preventing Melting enthalpy 12 11
misinterpretations. McCauley[76] describes the DSC in kcal/mol
curve of phthalylsulfathiazole, which presents such a
Water uptake after 0% 3.2% (Hydrate)
DSC curve with two endothermic peaks separated 1 day 92% RH
by an exothermic peak. The TG curve demonstrates
Transformation in alcohols A B ! A
a strong decomposition during the first melting.
 3736 Thermal Analysis of Drugs and Drug Products
Technology–
Tooling

Fig. 8 Reversible transition in a purine derivative studied by DSC and temperature-resolved X-ray powder diffraction. (From
Ref.[66].)

Above this temperature, the substance retains some of to the rubbery state and hence facilitating crystalliza-
the properties of a liquid, e.g., molecular mobility, and tion.[78] Since it may be interesting to maintain the
is termed ‘‘rubbery.’’ Above this temperature, the amorphous state, the temperature of the glass tran-
increase in molecular mobility facilitates spontaneous sition and its behavior should be characterized. DSC
crystallization into the crystalline form with an exo- and modulated DSC are commonly used. Hancock
thermic enthalpy change after the glass transition. and Zografi studied intensively the amorphous state
The use of amorphous forms is attractive, particularly of drug substances and used the relaxation energy at
for sparingly soluble compounds because of the the glass transition as well as the dependency of the
enhanced solubility and dissolution rate over the heating rate for the study of the ‘‘fragility’’ of the
crystalline state leading to increased bioavailability. amorphous state.[79–81] Depending on the temperature,
However, the amorphous state is thermodynamically the isothermal crystallization in one or the other poly-
unstable. The glass transition temperature Tg is low- morphic form may be favored as demonstrated for
ered by water or other additives, facilitating conversion amorphous indomethacine.[82]
Thermal Analysis of Drugs and Drug Products 3737

The study of the amorphous state (for drug sub-

stances and also for excipients) is based upon the 3
changes observed in the glass transition. Fig. 9 shows
the influence of the heating rate for the determination
of the glass transition by DSC for the polymeric excipi- 90.0 110.0 130.0
ent Carbopol 974. It is classical to perform a first run Temperature (˚C)
for the elimination of water and relaxation energy LOD (signal/noise ratio 3:1) is better than 1%
and to determine the glass transition temperature Tg
of the pure compound, if no degradation occurs during
Amorphous
the first run. However, this procedure is not valid if it is 2
desirable to study the role of the matrix.
When the amorphous material does not transform
into the crystalline material, the measurement of the

ENDO
melting peak allows to determine the degree of crystal-
linity of mixtures by comparing its value with the
melting enthalpy of a pure crystalline material. If
1
the amorphous sample crystallizes upon heating, then
the crystallization peak may be used for the determi- Crystalline
nation of the amorphous content. Such an example is
given in Fig. 10. In this case itwas possible to attain
a limit of detection of 1%. 75.0 125.0 175.0 225.0 275.0 ˚C

Technology–
Tooling
Fig. 10 Determination of amorphous content by DSC at
20 K min1. 1) Crystalline sample, 2) amorphous sample, 3)
Pseudo-polymorphism exotherm of a sample containing 4% amorphous as calcu-
lated by DSC. Calculation of the limit of detection by
In the case of solvates, binary phase diagrams of peak/noise ratio.
temperature versus concentration of the solvent (or
water) at a given pressure are useful for the under-
standing of the phase transitions. The characterization evaporation and possibly further endothermic or exo-
of solvates and hydrates need the use of both DSC and thermic events corresponding to a cascade of phase tran-
TG. Desolvatation can be complex: melting of the sitions. In such complex situations, combined techniques
solvate followed by exothermic recrystallization into TG-IR or TG-MS and temperature-resolved X-ray
the anhydrous form or solid-state transformation with diffraction are extremely helpful since the identity of the
volatile component or of possible volatile decomposition
product can be identified on line.[53,66] In case of hydrates,
5.0 ˚C/min
water sorption–desorption isotherms as well as the X-ray
diffraction in humid chambers are needed.[53,83]
The DSC, TG curves of solvates and hydrates are
related to the phase diagrams between substance and
solvent (or water). Eutectic are observed.[84] Fusion
20.0 ˚C/min or decomposition of the solvate may occur during
heating. Therefore, one may observe the melting of
the solvate followed by recrystallization into the anhy-
drous form or the endothermic desolvatation in the
40.0 ˚C/min solid state. In certain cases both phenomena may over-
lapp. Details about experimental factors and examples
can be found in Ref.[66]. If the anhydrous form is
metastable, further phase transitions follow the desol-
vatation. If several solvates or hydrates exist, the
transitions observed depend on the pressure, as
160.0 170.0 180.0 190.0 200.0 210.0
demonstrated by Soustelle[85] in the case of copper
sulfate pentahydrate. Depending on the pressure, the
Temperature (˚C)
direct dehydration into the anhydrous or the dehy-
Fig. 9 Quality control of excipients by using the glass tran- dration via the monohydrate, or the three dehydration
sition. Example of Carbopol 974 P. Second DSC run in order steps trihydrate, monohydrate and anhydrous forms
to eliminate water and relaxation: Influence of the heating rate. may be obtained. Hydrates have been the subject of
 3738 Thermal Analysis of Drugs and Drug Products

several reviews.[86,87] Polymorphism of hydrates[88] is Study of Transition: Kinetics

also frequent. Fig. 11 shows the case for a drug sub-
stance with polymorphic behavior of the trihydrate. Since transitions may occur during milling, processing,
The anhydrous form shows a dual melting (enantio- and aging, thermal methods are widely used for the
tropy). This form was very hygroscopic and trans- kinetic study of all transformations. The purpose of
formed into the trihydrate HI at room temperature. any kinetic study is to obtain information concerning
The DSC scan of this trihydrate shows a dehydration the reaction mechanism through comparison of a series
peak in solid state, followed by the same scan as on of measured fractions converted versus time. Most
the original anhydrous form. After storage for several mechanisms in solid state are a nucleation period, a
months in a tropical climate (30
C/75% RH), a second growth zone, and an unreacted core. For phase transitions
trihydrate HII is obtained. The same trihydrate HII is of polymorphs and pseudopoymorphs, only heterogene
also obtained by crystallization in water or after equi- kinetic applies (at least two modifications or two phases
libration of the original anhydrous form with saturated and a gaz). In heterogene kinetic a great number of factors
aqueous solution. The DSC scan of this trihydrate should be considered as temperature gradient in the
differs from the first one because the dehydration gives sample, particle size, activation, nucleus, or diffusion.
rise to a new anhydrous form. This interpretation was A summary of current kinetic methods used with
confirmed by X-ray Guinier de Wolff diagrams, purity thermal analysis techniques can be found in Ref.[66].
analysis, Karl Fischer, and TG.[10,66]
Often the solvates (hydrates) are not detected since,
according the corresponding phase diagram, at ambi- Microcalorimetric Techniques
ent temperature, they can be partly or completely
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dissociated. Suspensions of hydrates in water should Solution calorimetry allows us to investigate processes
Tooling

shift the equilibrium toward the formation of the stable that involve enthalpy changes. Adiabatic microcalorim-
hydrated form. The ability of DSC measurements at eters and isoperibol calorimeters used in batch
subambient temperatures allow to determine phase modes or flow modes allow for the precise determi-
transitions. Giron et al.[89] proposed to use the melting nation of the heat of solution. Mixing the reactants is
peak of freezable water for the analysis of suspensions accomplished by breaking a bulk allowing reactants
of drug substances in water in combination with TG to mix or by special chambers where the reactants
for the determination of the number of molecules of are mixed together.
water bounded as hydrates. If a compound exists in two or more different crystal-
line or amorphous configurations with different lattice
energies, the heating solution in any given solvent will
differ. The difference in the heats of solution will be equal
II to the difference in lattice energy of the solids, provided
that the solid compounds are identical chemically. For
example, we measured the energy of solution in water
of the two modifications of a drug substance. The differ-
ence of 9.7 kJ mol1 was found very close to the differ-
ence of the melting energies of 9.1 kJ mol1 measured
Endothermic

by DSC.[53] However, the DSC information is superior

since the temperatures of melting of both forms are
–H2O measurable. Since the lower melting form had a lower
I melting energy, it was the stable form and both forms
were enantiotropically related. For review see Ref.[66].
Byström[90] developed a technique in order to deter-
mine the crystallinity of drug substances by isothermal
microcalorimetry. Considering that micronization intro-
–H2O duces amorphous regions not measurable by X-ray
diffraction, the method should be an analytical tool for
analyzing batch-to-batch quality. The principle of the
measurement lies in the transformation of the amorphous
state in the crystalline one at high humidity levels. The
70 80 90 100 110 120 130 140 150 160 ˚C amorphous substance adsorbs water and the glass tran-
Temperature (˚C)
sition is lowered, permitting the acceleration of the crys-
tallization. The energy evolved is measured in function
Fig. 11 Polymorphism of a trihydrate. (From Ref.[10].) of time by isothermal microcalorimetry. Results were
Thermal Analysis of Drugs and Drug Products 3739

found comparable with X-ray diffraction but a quite

T dQ
lower limit of detection is possible (better than 1%).[53,91,92] dt
Liq. Tm
Tm To
DSC Purity Analysis
A + Liq. B + Liq.
TE
The basis of any calorimetric purity method is the A+8
relationship between the melting depression of a sub- 0% X2 100% A TE Tm
stance and the level of impurities according to van’t
Fig. 12 Binary phase diagram with eutectic, and DSC curve
Hoff’s law. The purity is readily calculated from the
of the composition x2.
DSC curve of a single melting event of a few milli-
grams of the substance, without the need for reference
standard of the drug substance and its impurities. solution formation, the concentration of impurity in
The DSC impurity analysis is described in USP. the liquid phase at any temperature during the melting
With modern equipment including robotic systems is inversely proportional to the fraction melted at that
and data aquisition, the DSC purity analysis is a state- temperature, and the melting-point depression is
of-the-art technique for pharmaceutical development. directly proportional to the mole fraction of impurity.
The determination of purity by means of DSC is A plot of the observed analyte temperature Ti vs. the
based on the assumption that impurities depress the reciprocal of the fraction melted 1/Fi at temperature
melting point of a pure material according to the eutec- Ti should yield a straight line with the slope equal to
tic phase diagram behavior. the melting-point depression (T0  Tm). The theoreti-

Technology–
Fig. 12 shows the phase diagram for the two compo- cal melting point of the pure compound T0 is obtained

Tooling
nent mixture with the so-called eutectic point. At the by extrapolation to 1/Fi ¼ 0:
eutectic point E (e.g., 40% A, 60% B), the crystals A
and B melt together at the temperature TE, below the RT02 ð1=Fi Þ
Ti ¼ T0  x
melting temperature of the pure compounds. If a mix- DHf
ture of A and B (containing, e.g., 90% A) is heated, the
melting of eutectic mixture (which is 40% in A) is This relation may be expressed as
observed initially, until all of B is melted. During the
melting of the eutectic (40% A, 60% B) a part of A Ti ¼ T0  DTð1=Fi Þ
is melted with B, with the corresponding amount
2/3  10% of A, i.e., 6.66% of A. Substituting the experimentally obtained values for
Then as the temperature increases, pure A melts DT, DHf, and T0 in the first equation yields the mole
between TE and Tm. Tm is the temperature at the end fraction of the total eutectic impurities, which, when
of the melting. For the corresponding DSC curve, an multiplied by 100, gives the mole percentage of total
endotherm at the eutectic temperature is observed, eutectic impurities.
then the melting of crystals A occurs. The effect of
impurity on the DSC curve is a melting depression (PSA)0,0%
and a broadening of the melting curve (Fig. 13).
The amount of impurities is calculated from the (DSC)0,8%
(DSC)0,7%
melting-point depression DT ¼ T0  Tm.
The van’t Hoff’s law for diluted solutions is
1.25 ˚C.min–1
(PSA)1,7%
ðDT DHf Þ
x ¼ (DSC)1,5%
RT02
Endo

(PSA)5,5%
where x is the mole fraction of impurities, DT the melt-
ing point depression, DHf the melting point of pure
material, Tm the melting of the analyte, T0 the melting
point of the pure compound, and R the gas constant.
126 128 130 132 134 136 ˚C T
The DSC procedure does not directly measure DT,
but can be used to calculate it from the melting curve. Temperature
At the eutectic point, all of B is in the liquid phase. Fig. 13 Broadening effect of the melting curve of b-hydroxy-
During the melting of A after the eutectic point the propyltheophylline due to impurities. PSA ¼ phase solubility
concentration of B varies in the liquid phase. This analysis. All batches have the same TLC purity results. (From
causes the broadening of the DSC curve. With no solid Ref.[6].)
 3740 Thermal Analysis of Drugs and Drug Products

The temperature of each point Ti is the sample  The sum of impurities should be 2%.
temperature, not the programmed temperature. Due to  The result is expressed in mol % without knowledge
the thermal lag, a correction depending on the instru- of impurities.
ment has to be done for each point.  Pure material is not needed.
The melting curve is divided into small portions and  Small amounts (1–2 mg or less) of material are used.
each area Si is calculated. The melted fraction Fi,  If decomposition occurs during melting, it can give
erroneous results.
Si  The purity results are obtained after less than 1 h.
Fi ¼
Stotal
The influence of products parameters and instru-
is calculated for each point and the curve Ti is plotted ment parameters have been discussed in detail by
as a function of 1/Fi, where Ti is the temperature at Giron and Goldbronn,[93] who proposed a validation
fraction Fi (Fig. 14). The slope DT and the ordinate scheme for DSC purity method. The authors showed
T0 can be calculated. the advantage of the method as support to chromato-
Partly because of the lack of the eutectic-point graphic techniques, for the monitoring of purification
detection, the curve is not a straight line, and a correc- for the study of stability behavior of raw materials
tion factor K must be added to each fraction of the under stress conditions, for establishing purity profiles.
curve. Formation of solid solution or artefacts during Fig. 15 shows the DSC scans corresponding to a
melting may also be responsible. stability screening of a drug substance sensitive to
moisture.
Si þ K
Fi ¼
Technology–

Stotal þ K
Tooling

Enantiomers
Software from manufacturers mostly use iterative line-
arization, which gives the best value of K. Characteris- Enantiomers are stereoisomers, which are mirror
tics of this determination are as follows: images of eath another. An equimolecular mixture of
two enantiomers is called a racemate. Crystalline race-
 Impurities are measured, which have an eutectic mates occur in three different types. The first is termed
behavior (i.e., solubles in the liquid phase and inso- a conglomerate, that is, a mechanical mixture of crys-
lubles in the solid phase). tals of pure enantiomers that is formed from two solid
phases. The most common type is the racemic com-
pound, which consists only of one crystalline phase
in which the two enantiomers are present in equal
Endo Temperature
A
correction quantities. The third type is the pseudoracemate in
which a solid solution of the two enantiomers is
present.

84.0
B 82.0
104 105 106 107 ˚C 108 99.9%
TAcorr. 80.0
99.7%
Heat flow (mW) –>

78.0
76.0
x = – ∆T . ∆H
Si
F= 74.0
R . To2 ST 99.1%
72.0
378
T/K

ST + K 70.0
1/F =
K = 8% ST Si + K 68.0
66.0
K = 8% ST ∆T = –0.35
377 T o = 378 K 64.0
112.0 114.0 116.0 118.0 120.0 122.0 124.0 126.0
x = 1.0% Temperature (˚C) –>
K=0
Fig. 15 Stability screening. Example of DSC purity results
376
0 4 6 8 obtained for a drug substance sensitive to moisture. From
1/F the top to the bottom: Initial sample (purity 99.9%), sample
stored under humid conditions at 50
C (purity 99.7%), sam-
Fig. 14 Purity calculations by DSC. ple stored under humid conditions at 80
C (purity 99.1%).
Thermal Analysis of Drugs and Drug Products 3741

Conglomerates that are equimolecular mixtures of Phase diagrams with monotropic transformation or
two crystalline enantiomers are easily separated by enantiotropic transformation have been discussed.[74,75]
cristallization. There are two phases in the solid state Quite interesting is the transformation of the racemic
and only one phase in the liquid state (miscibility). compound into a conglomerate since this phenomenon
The equation of Schröder–Van Laar in its simplified can be used for purification via crystallization, as
form correlates the composition of mixtures to the described for nimodipine.[96] DSC is applied for the
end of fusion Tf: establishment of phase diagrams, for the determination
of thermodynamic data,[97] for the purity determi-
  nation, or for the monitoring of industrial resolutions.
DHAf 1 1
ln x ¼  f For the establishment of phase diagrams it is suitable
R TAf T
to add spectroscopic or crystallographic methods.[98]
Two methods can be used for the purity determi-
where x is the mole fraction of the more abundant nation: the direct method or the indirect method.[74]
enantiomer (0.5  x  1) of a mixture that melts at The direct method is applied for mixtures when the
Tf (in K). DH fA and T fA are respectively the enthalpy phase diagram is established using Schröder–Van Laar
of fusion and the melting point of the pure enantio- or Prigogine–Defay equation. For enantiomers of high
mers, and R is the gas constant. purity (>95%) the general DSC purity method for
Racemic compounds, or true racemates, exhibit two eutectic impurities is applicable. The same limitations
eutectic points each between the pure enantiomer and remain (polymorphism, degradation, during melting).
the racemic compound. The shape of the DSC dia- The method gives the sum of impurities without differ-
grams can vary, depending on the relative positions entiation of the type of impurity. For purity >95% the

Technology–
of temperatures of eutectics and racemic compound same results have been obtained using the indirect and

Tooling
and on the composition of the eutectics (Figs. 16B the direct methods.[99,100]
and C). In the case of Fig. 16C it is difficult to distinguish Conglomerates are easy to purify by crystallization
by DSC a racemic compound from a conglomerate. or by the entrainment technique described by Jacques
Other methods as IR or X-ray are suitable for proper et al.,[74] which involves introducing seed crystals of
interpretation. For example, propanol hydrochloride the desired enantiomer into a cooling saturated solu-
has been described as conglomerate[94] or as racemate tion of the racemic mixture. According to these
compound.[95] authors, conglomerate formation is observed three
The equation of Schröder–Van Laar, which permits times, more frequently with salts. Therefore it is advan-
the calculation of the liquidus curve, may be applied to tageous to compare the behavior of salts forms. On the
the point of the liquidus between the pure enantiomers other hand, for a synthesis in several steps, it is useful
and the corresponding eutectics (TfAER and TfAES. For to study the behavior of each step in order to choose
the part ERRES, the equation of Prigogine and Defay the step exhibiting a conglomerate behavior resulting
applies: in an efficient enantiomeric resolution. In the case of
  racemic compound, the entrainment technique can be
2DHRf 1 1 used in those cases where the eutectics are situated very
ln 4xð1  xÞ ¼  f
R TRf T close to the racemate.This underscores the value of sys-
tematic searches for derivatives that form conglomer-
where x represents the mole fraction of the enantiomer ates or at least racemic compounds whose eutectics
in the mixture that melts at T f and DH fR and T fR are are close to that of the racemate.
respectively the enthalpy of fusion and the melting Another further way of purification is the formation
point of the racemic compound. Polymorphism can of diastereomeric salts. However, partial solid solution
occur for the racemic compound and the enantiomers. are often observed and it is difficult to achieve high

A B C D
TA
TR 2
TA TA TA
T
TE TR 1
TE
TE TE 3

x% 50% XE 50% XE

Fig. 16 Phase diagrams for enantiomers: (A) conglomerate, (B) and (C) racemic compound, (D) solid solution.
 3742 Thermal Analysis of Drugs and Drug Products

purity. Resolution via diastereomeric salt formation The thermal analysis studies of different forms of
based on a part of the DSC curve has been discussed.[101] sorbitol, mannitol, glucose, magnesium stearate are
The change from racemates to enantiomers has reviewed in Ref.[67]. Mono-, di-, and trihydrate of mag-
implication in the galenical formulations. Propanolol- nesium stearate as well as the amorphous form may
base enantiomer has a better skin permeation than be found. According to Wada and Matsubyara[109]
racemate, what was explained by a lowering of the thermal analysis is the most appropriate method for
melting point.[102] The stereospecific preformulation characterization of magnesium stearate.
of ibuprofen has also been discussed. The theoretical Thermal analysis are widely used for polymers and
phase diagram has been calculated from the DSC copolymers analysis.[4] Glass transitions, melting, and
curves of the (þ) enantiomer and of the racemate decomposition processes are analyzed. Since the glass
by using the equations of Schröder–Van Laar and transition temperature Tg is marked by changes in
Prigogine–Defay equations. Experimental data con- the thermal capacity, expansion coefficient, and rigid-
firm the calculated data.[103,104] The low melting point ity, TMA technique as well as DSC may be used. Tg
of the enantiomer may have implications of the phar- increases with molecular mass up to certain values.
maceutical process. Relationships between physical Plasticizers and water depress this temperature. Ther-
properties and crystal structures of chiral drugs have mal stability and influence of antioxidants and fillers
been discussed.[105] may be analyzed by TG or DSC, under oxygen.
The compatibility of polymers in blends is tested by
comparing DSC curves of the components. Immiscible
crystalline blends such as polyethylene–polypropylene
APPLICATION OF THERMAL TECHNIQUES FOR show the DSC peaks of polyethylene and poly-
Technology–

THE DRUG PRODUCT: PREFORMULATION, propylene. Immiscible amorphous blends exhibit two
Tooling

PROCESSING, AND AGING glass transitions; in miscible blends a new glass point
is observed. Partially miscible blends have two glass
The preformulation includes the choice of the salt form points situated between the glass points of each poly-
and of the polymorph of the drug substance. Melting mer. Different equations such as Gordon–Taylor
points, solubilities, dissolution, hygroscopicity, sta- express the relation between the new glass transition
bility, feasibility, processability, and polymorphic behav- Tg and the glass transition points Tg1 and Tg2 of com-
ior have to be considered. The second step is to study the ponents. W1 and W2 are the weight fractions and K is
behavior of the drug substance with excipients.[106] the ratio DCp2/DCp1.

Tg ¼ W1 Tg1 þ KW2 Tg2 =W1 þ KW2

Excipients The polymers mostly used in pharmaceutical pack-

aging are polyethylene, polypropylene, PVC, poly-
The use of thermal analysis techniques for pharma- amide, polystyrol, nylon, cellulose acetate, polyethylene
ceuticals implies the knowledge of the thermal behavior terephthtalate, and blends thereof. Copolymers and
of the excipients. A great number of publications deal rubbers are also used. The DSC melting curve of poly-
with polymorphic behavior of excipients and especially ethylene used for packaging purposes is characteristic.
with the amorphous forms as prerequisite knowledge Low- and high-density polyethylene are differentiated
for freeze drying and milling processes. by their melting points.[110] Melting point and density
Lactose exists in two isomeric forms a and b. It is of polyethylene are linearily correlated.[111] Crystal-
possible to obtain the a-monhydrate, anhydrous linity may be determined as described above for
crystalline a and b forms, as well as an amorphous amorphous state.
form. Thepharmaceutical properties of these various Polyethylene glycols (PEG) have been intensively
types are different. The hardness of tablets obtained characterized by thermal analytic methods. The melt-
using amorphous lactose produced by lyophilization ing points of PEG increase with the molecular weight
is 10 times that obtained using crystalline forms. and decreases with the content of water as a result of
During milling, it has been observed that the mono- eutectic formation.[10] Corrigan[112] studied the differ-
hydrate loses part of its water of crystallization and ent DSC peaks of PEG: Once folded chain crystals
of its crystallinity. Heat treatment of different lactoses and extended chain crystals are present in PEG 6000,
has permitted the discovery of an anhydrous, unstable which results in two DSC peaks. In PEG of higher
a form and a crystal containing a and b in the ratio molecular mass, only folded chain crystals are present.
1 : 1. Under the influence of high degrees of humidity, Lower molecular mass PEG contain only extended
amorphous lactose crystallizes and anhydrous forms chain crystals. Craig reviewed the thermal studies of
tend to reconvert to the monohydrate.[67,107,108] PEG, including properties in aqueous solutions.[113,114]
Thermal Analysis of Drugs and Drug Products 3743

Phase diagram of PEG 4000 was recently discussed.[115] 8.0

Higher molecular mass PEG are generally used in solid Melting
dispersion systems and their melting behavior is relevant
for the temperature of galenical preparation.[5,116]

Endo>
Glass transitions of polyvinylpyrrolidones of differ-
ent molecular weight may be used as identity.[10,117]
Water depresses the glass transitions and effects the

Mcal/sec
physical properties of polyvinylpyrrolidone, as demon- 4.0
strated by Tan and Challa.[118] Glass transition Crystalisation
A large number of cellulose derivatives have been
studied for food as well as for pharmaceutical applica-
tions.[119–122] The measurement of the glass transition
is often difficult due to the broad endotherm of
dehydration of water. Fig. 17 shows the effect of water
on the depression of the glass transition of hydroxy- 0.0
propyl methylcellulose HPMC4000. 50 90 130 170 190
Film coating processes need the knowledge of Temperature (ºC)
the glass transitions for a proper film. Ethylcellulose,
Fig. 18 DSC curve of L-polylactic acid after quenching from
cellulose phthalate, polyvinylalcohols, polymethyl
the melt. (From Ref.[10].)
methacrylates have been studied and the critical param-
eters for film formation discussed.[123–125]

Technology–
Polysaccharides and water interaction are especially of the drug substance to be analyzed; thermal techni-

Tooling
studied for their use in spray-dried products in food ques generally with electron microscopy are used for
and in biotechnology.[126–128] the optimization of the drug loading and of the pro-
Biodegradable polymers can be crystalline or cess.[131] The physical aging of the polymer can be
amorphous. Poly-L-lactic and D,L-polyactic acids have assessed in DSC by the amplitude associated with the
been studied by Pitt and Gu.[129] Fig. 18 deals with glass transition of the matrix. This relaxation energy
the second run of poly-L-lactic acid after quenching. increases with aging. Rosilio studied the progesterone
The glass transition is followed by crystallization, the poly(D,L-lactide-co-glycolide) microspheres. Aging acts
melting of the crystalline form. Aging and crystallinity on the solid state of the drug substance loaded. Different
of biodegradable polymers have been studied by polymorphs are obtained, depending on the copolymer
Akktar et al.[130] TG is useful for the determination of composition.[132]
the entrapped solvent (often methylene chloride) and
of moisture.[5,10] Microspheres containing biodegrad-
able polymers are intensely analyzed by thermal Phase Diagrams
analysis techniques allowing both the glass transition
point of the polymer and the physico-characterization The thermodynamic phase diagrams are the basis for
understanding DSC curves of formulation: eutectics,
solid solutions, eutectics with partial solid solutions,
130 and compound formation with congruent or incongru-
120 ent melting. Fig. 19 exemplifies the building of the
110 phase diagram of propyphenazone and butesamide,
Temperature (˚C)

using the DSC curves. In order to save the number

100
of DSC scans, theoretical curves can be added.[133]
90
Such phase diagram was performed between a drug
80 substance and stearic acid: no formation of the salt
70 was observed.[12] Recently thermal analysis has been
60 used for the complex phase diagram of propanol/oleic
acid for which the salt and a mesomorphic phase have
50
been found.[134]
40 DSC was proposed for compatibility studies com-
0 2 4 6 8 10 12 14 16
paring DSC curves of components and mixtures.
% Water
Unfortunately some misinterpetation may occur. DSC
Fig. 17 Influence of the water content on the glass transition curves can only reflect physical behavior. The forma-
temperature of HPMC 4000 measured by DSC in sealed tion of eutectic is not an incompatibility. Furthermore,
pans. water generally is not present in the mixtures at the
 3744 Thermal Analysis of Drugs and Drug Products

Heat eutectic
40%
100%

10%

100 80 60 40 20 0%

Butesamid
Propyphenazon

Propyphenazon
95%
Endotherm

T T
(°C) (°C)

100
90%
80% 90
80%
80
90% 70
95%
60

60 70 80 90 100 T (°C) 0 10 20 30 40 50 60 70 80 90 100

Technology–

Temperature (°C) Propyphenazon (%)

Tooling

Fig. 19 Example of building a phase diagram by DSC of propyphenazone and butesamide. (From Ref.[6].)

temperatures of the melting peaks. Giron[9] performed preparation, the type of systems, the dissolution
the DSC curves of a drug substance and several excipi- characteristics, and the aging problems should be
ents, the initial DSC curves of the mixtures and the considered. DSC is appropriate for the study of solid
DSC curves of the mixtures after 1 month at 50
C/ dispersions by comparing DSC curves of pure com-
<30% RH and 50
C/75% RH. Chromatography was pounds, of physical mixtures or melted mixtures to
used for the chemical analysis of the drug. Fig. 20 the DSC curve of the solid dispersion. TG, X-Ray dif-
shows the comparison of the DSC curves of the mix- fraction, and hyphenated techniques are good comple-
tures. The effect of the humidity on the degradation ments. The transitions observed are those transitions
is very striking: The drug substance is completely expected from the phase diagrams, or the amorphous
decomposed with calcium sulfate, avicel, and stearic state with glass transition or very often new metastable
acid. For talc and dicalcium phosphate, there is no forms of drug substance as well as metastable forms of
degradation. The observation of the initial DSC curves excipients.
for the five excipients would have wrongly given the The phase diagram given in Fig. 21 for darodipine–
conclusion to the same compatibility. polyethylene glycol 6000 results from DSC experi-
For such compatibility studies, isothermal micro- ments carried out by two techniques.[10] The DSC of
calorimetry has been suggested (e.g., Ref.[135]). physical mixtures obtained by grinding were scanned
The DSC study of mixtures of drug and excipients is just after the end of melting. After cooling a second
very useful, for the information gained (e.g., if the scan at 5 K min1 was performed. In the second tech-
eutectic melts at ambient temperature). In other cases, nique the mixtures were dissolved in methanol and
one may target interaction with excipients as it is the the solvent evaporated. With both techniques, the same
case of solid dispersions, solid solutions, or complex results were obtained.
formation. The choice of the carrier defines the charac- The knowledge of the phase diagram allows the
teristics of dissolution of the dispersed drug. Poorly choice of the temperature of the process. It can be suit-
water-soluble active ingredients are combined with able to choose higher loading than the eutectic compo-
water-soluble carriers in order to increase the dissolu- sition. Once the solid dispersion manufactured and
tion of the active ingredient. A good water-soluble milled, DSC is very advantageous for checking the
drug combined with a slightly soluble carrier leads to batch reproductibility or the stability behavior by the
a retardation of drug release from the matrix. For measurement of the melting enthalpy of the eutectic.
reviews on solid dispersions, see Refs.[5,136]. For For a solid dispersion with 40% darodipine, a standard
the development of solid dispersions, the method of deviation of 1.4% for the melting point of the eutectic
Thermal Analysis of Drugs and Drug Products 3745

1 Fresh mixture
2 After 1 month 50ºC
3 After 1 month 50ºC. 75% r.h.

Lactose Talc

1
1
3 2 3

Mg-Stearate CaSO4· 2H2O

2 2
3
1 3 1

Technology–
Tooling
Mannit Avicol
2
3

1
2 1 3

Stearic acid
Dicaphosphate

2 2
1
1
3

50 100 150 190 50 100 150 170

(ºC) (ºC)

Fig. 20 DSC curves of 10% drug substance with excipients: 1) Original curves; 2) curves after 1 month at 50
C/<30% RH; and
3) 50
C/75% RH. The initial DSC curves suggest an incompatibility only with mannitol and magnesium stearate. The DSC
curves (curves 3) after storage at 50
C/75% RH allow the differentiation of the excipients: degradation with stearic acid, calcium
sulfate dihydrate, and avicel.

located at 58
C and 2.4% for the heat of eutectic of Solid solutions: The drug and the carrier are
27 cal g1 was found. miscible in the solid state. Polyvinylpyrrolidones (PVP)
PEG used for solid dispersion vary from molecular with different molecular weights dissolve drug sub-
weight 1000 to molecular weight 20,000. PEG of lower stances such as diazepam[137] through hydrophobic
molecular weight are liquid and therefore not suitable. interactions. The disparition of the DSC peak of the
The quality mostly used are PEG 4000 and PEG 6000. drug substance demonstrates the formation of the solid
The composition of the eutectic of most drug sub- solution.
stances lies in small concentration of drug substance. Drug substance and carrier can be both in the
Some monotectics (0% drug substance) have been glassy state. Total miscibility, partial miscibility, or
described. The highest amount of drug is obtained with total immiscibility has been observed. This type of
gluthetimide with an eutectic composition of 32%.[5] system is very sensitive to temperature and moisture
 3746 Thermal Analysis of Drugs and Drug Products

for the monitoring of the dissolution characteristics

of the drug. These cyclic oligosaccharides contain six
160
(a-CyD), seven (b-CyD), and eight (g-CyD) a-(1,4)-
linked glucose units. A great number of chemically
modified cyclodextrins have been manufactured in
140
the last decade. Complexes are formed through
inclusion in the cavity or through interactions with
chemical groups. In order to obtain complexation,
120
the compounds have to bind a complex first in solu-
tion. From the usual methods of preparation, kneading,
coprecipitation, freeze drying, or spray drying, often
the spray-drying technique gives the best results.
100
Giordano, Bruni, and Bettinetti[139] suggested a
method of calculation of the ratio guest/host. They per-
formed DSC analysis of dispersions of different compo-
80 sitions with an excess of the guest molecule. The
remaining energy in the melting peak of the guest mol-
ecule allows the calculation of the amount of free drug.
60 They plot this amount for different composition versus
the total guest fraction and compared the plot obtained
with the theoretical plots for ratios 1 : 1, 1 : 2, 1 : 3.
Technology–

0 10 20 30 40 50 60 70 80 90 100 In most cases the DSC peak of the drug substance

Tooling

% Drug substance disappears and no new peak is observed. With hydro-

cortisone butyrate,[140] a new peak corresponding to
Fig. 21 Phase diagram of a solid dispersion of darodipine–
the complex has been observed. For this drug, the
PEG 6000.

and crystallizations are often observed after long

storage. Batch 1
New derivatives of starch, mainly cyclodextrins[138]
which forms inclusion compounds, are best carriers
3 months 30˚C
Scan rate 20.00 deg/min

1 year 25˚C
Raw material

3 months 25˚C

Solid fraction
deep-freezer

˚C
40.0 50.0 60.0
After 1 year the same DSC curve is obtained at 25 ˚C, 30˚C as after
2 years as demonstrated with another batch.

Liquid
Batch 2 2 years 25˚C
deep-freezer

240 260 280 300 320 340 360

Temperature (K)
40.0 50.0 60.0
Fig. 22 DSC curves for the monitoring of fractionation of a
liquid excipient. Scan rate 20 K min1. Fig. 23 Aging of precirol.
Thermal Analysis of Drugs and Drug Products 3747

complexation increases in the order a-CyD, b-CyD,

g-CyD, and dimethyl-b-CyD. Thermogravimetry analysis T T
has also been used.[141] The temperatures of vaporization, Liq.
sublimation, or degradation of the drugs are displaced to
higher temperatures, due to the complexation.

Analysis of the Drug Product

Solution

If the components are not miscible in the solid and the

liquid states, their DSC peaks remain unchanged. This Rubber
allows to identify components of the drug product, to
E
follow aging problems with polymorphism and in
favourable cases to quantify the drug substance and Ice + sol.
the excipients.[20,142,143]
Glass
Fatty Acids and Glycerides Derivatives
Tg
Most fats and similar compounds show polymorphism
behavior, including fatty acids, fatty alcohols, ceto-
stearyl alcohols, glycerides, oils, hydrogenated or trans- 100%H2O 100% B
esterified oils, and suppositories.[144] This polymorphic

Technology–
Tooling
behavior is characterized by change of melting by Fig. 24 Phase diagram of water and amorphous substance
during freeze drying.
aging, giving rise to hardning effects of suppositories
and precipitations of liquid excipients that are complex
mixtures of glycerides. Examples of aging problems are acid derivatives are unsaturated, they are very sensitive
given in Ref.[10]. Fig. 22 is an example of the control of to oxidation. The oxidability and the influence of anti-
the fractionation of a liquid excipent, using DSC. Aging oxidants can be measured by DSC and TG, comparing
of the precirol excipient is demonstrated in Fig. 23. Sub the starting of the degradation.[147]
ambient DSC was also used for the preformulations of
microemulsions.[11–12] The solid fat index, calculated as Interaction with Water
a percentage of the solid as function of the temperature,
is very valuable for the evaluation of suppositories.[145] The phase diagrams of drug substance and excipients
Analysis of the drug is also possible. Chemical reaction with water as well as the study of the temperature of
was observed for aminophylline.[146] Since most fatty glass transition (Fig. 24) are the basis of the choice

A B C
Freezable water in a gel

Cetylpalmitate with spiked water

Endotherm

Methocel K 15 M
with spiked water

Water

Methocel K 15 M alone

0.0 25.0 50 0.0 10.0 20.0 30.0 –20.0 0.0 20.0

Temperature (°C) Temperature (°C)

Fig. 25 Determination of freezable water, using the melting peak of ice. (A) Methocel K15 M, (B) cetyl palmitate, and (C) phar-
maceutical gel.
 3748 Thermal Analysis of Drugs and Drug Products

of the conditions of freeze-dried or spray-dried formu- 3. Wunderlich, B. Thermal Analysis; Academic Press: New
lations.[148–151] The lyophilizates are well characterized York, 1990.
4. Turi, E.A. Thermal characterization on polymeric materi-
by DSC and TG. The polymorphic behavior of all als, 2nd Ed.; Academic Press: New York, 1997.
components can be studied by thermal techniques. 5. Ford, J.L.; Timmins, P. Pharmaceutical thermal analysis
For proteins, it is suitable to have excipients in the for- techniques and applications. In Series in Pharmaceutical
Technology, Ellis Horwood Books in Biological Sciences;
mulation that remains amorphous. Trehalose was Rubinstein, M.H., Ed.; John Wiley & Sons: New York,
found to be a very efficient lyoprotectant.[152] 1989.
Freezable water is determined by the measurement 6. Giron-Forest, D. Thermoanalytische Verfahren. In Phar-
mazeutischer Qualitätskontrolle; Feltkamp, H., Fuchs,
of the melting peak of ice. This technique is currently P., Sucker, H., Eds.; Stuttgart, Leipzig, Germany, 1983;
applied for creams, and gels.[153,154] Examples of the melt- 298–310Georg Thieme Verlag.
ing peak of freezable water are given in Fig. 25. Swelling 7. Barnes, P.A. Applications of new methods and instrumen-
tation in thermal analysis. Thermochim. Acta 1987, 114,
properties of modified release matrices and their inter- 1–8.
action during dissolution have been studied.[155,156] 8. Brennan, W.P. Some applications of thermal analysis as a
Monoglycerides–water systems have been character- supplement to or replacement for ASTM testing stan-
dards. Thermochim. Acta 1977, 18, 10–13, and 101–111.
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hexagonal) have been determined. Water/oil systems ceutical industry. J. Pharm. Biochem. Anal. 1986, 40,
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10. Giron, D. Thermal analysis in pharmaceutical routine
studied using DSC.[158,159] Thermogravimetry can be analysis. Acta Pharm. Jugosl. 1990, 40, 95–157.
successfully used additionally.[160] 11. Giron, D. Thermal analysis, microcalorimetry and com-
Liposomes are multilayered vesicles consisting of bined techniques for the study of pharmaceuticals. J.
Therm. Anal. Calorim. 1999, 56, 1285–1304.
concentric bilayers of phospholipids interdispersed 12. Giron, D. Contribution of thermal methods and related
Technology–

with aqueous phases. In aqueous media, the phospho- techniques for rational development of pharmaceuticals.
Tooling

lipids undergo gel, liquid crystalline transitions easy to P.S.S.T. 1998, 1, 191–262.
13. Cheng, S.Z.D.; Li, C.Y.; Calhoun, B.H.; Zhu, L.; Zhou,
detect by DSC. The study of the change of these transi- W.W. Thermal analysis: the next two decades. Thermo-
tions, temperature, peak width, and energy allows to chim. Acta 2000, 355, 59–68.
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mochim. Acta 2000, 347, 9–13.
16. Sarge, S.M.; Hohne, G.W.H.; Cammenga, H.K.; Eysel,
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bration of scanning calorimeters in the cooling mode.
Thermochim. Acta 2000, 361, 1–20.
CONCLUSIONS 17. Sabbah, R.; Xu-wu, A.; Chickos, J.S.; Leita, M.L.; Roux,
M.V.; Torres, L.A. Reference materials for calorimetry
Thermal analysis methods are widely used in all fields and differential thermal analysis. Thermochim. Acta
1999, 331, 93–204.
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22. Hohne, G.W.H. Modulated DSC. Thermochimica Acta
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23. Coleman, N.J.; Craig, D.Q.M. Modulated DSC: a novel
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 Titrimetry
Vesa Virtanen
Department of Pharmaceutical Product Development, Orion Pharma,
Kuopio, Finland

DEFINITION OF TERMS the concentration of the insoluble salt AB may be

regarded as being constant:
Titrimetry or titrimetric analysis is any method of
quantitative chemical analysis in which the amount Kppn½AB ¼ ½A
½Bþ  ¼ constant ¼ SAB
of a substance is determined by measuring the volume
that it occupies or the volume of a second substance where the constant S is the solubility product. If the
that is needed to react completely with the substance solution is pure, equivalent concentrations of A

being determined. Titration is a process of chemical and Bþ will be present, and therefore:
analysis in which the quantity of some constituents
of a sample is determined by adding to the measured ½A
 ¼ ½Bþ  or ½A
2 ¼ ½Bþ 2 ¼ SAB :
sample an exactly known quantity of another sub-
stance with which the desired constituent reacts in a In this case, if the product [A
][Bþ] exceeds (SAB)0.5,
Technology–

definite, known proportion. The reagent of exactly

Tooling

the solution is saturated with respect to AB, and this

known composition used in a titration is called a stan- substance separates as a precipitate.[1]
dard solution.
The goal of titration is the equivalence point, where
the addition of standard solution in an amount that is
chemically equivalent to the substance with which it Titration Curves
reacts. In fact, its position can be estimated only by
observing physical changes associated with it in the Titration curves are based on the negative logarithm
solution. These changes manifest themselves at the (to the base 10) of the molar concentration of the
endpoint of the titration. species; p-values or p-functions are useful for deducing
An indicator is a supplementary chemical compound the properties required of an indicator as well as the
that exhibits a change in color as a result of concentration titration error that its use is likely to cause.
changes occurring near the equivalence point. The general shape of a titration curve for the pre-
cipitation titration of a solution containing one anion,
Cl
, is shown in Fig. 1. Curves plotted from calculated
pX values are always symmetrical. A curve plotted
PRECIPITATION TITRATIONS by using experimentally obtained values for [Cl
] is
not symmetrical. The lack of symmetry is due to those
In these titrations, the determination of the substance ions that are being adsorbed in excess by the precipi-
is effected by precipitating it in the form of an ‘‘insolu- tate in different amounts. This fact is not taken into
ble’’ compound of known composition. The equiva- account in theoretical calculations. Accurate precipi-
lence point is reached when sufficient reagent to tation titrimetry requires a long, steep AB section because
complete precipitation has been added. In practice, the length of AB depends on the initial concentrations of
the insoluble reaction product formed will be very the reactants and the value of the solubility product of
slightly soluble and to an extent that depends on the the compound precipitated. An increase in analyte and
amount of solvent present as well as on the nature reagent concentration enhances the change in pX in the
and amounts of other ions and compounds present. equivalence point region.
In the simplest general case of a slightly soluble salt Precipitation titrations can be extended to mixtures
(AB) formed by the reaction of the oppositely charged that form precipitates of different solubilities. The
univalent anion (A
) and cation (Bþ) of two soluble salts: titration curve in Fig. 2 for a chloride/iodide mixture
is a composite of the individual curves for the two
A
þ Bþ ¼ AB and Kppn ¼ ½A
½Bþ =½AB anionic species. Because silver iodide has a much lower
solubility than does silver chloride, the initial additions
where Kppn is the precipitation constant. Assuming of the reagent result exclusively in formation of iodide.
that interferences are small enough to be neglected, Thus, two equivalence points are evident.
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-100000442
3752 Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved.
Titrimetry 3753

produces a visually detectable change in the solution,

usually color or turbidity, or may form another
A
precipitate that has a distinctive color. The indicator
functions by reacting competitively with one of the
reactants or products of the titration and, therefore,
pX

its concentration must be kept low, favoring an intense

Centre of AB color at the endpoint. For some determinations, an
electrochemical sensor provides an accurate way of
locating the equivalence point.
B The formation of a second, highly colored precipi-
tate is the basis of the Mohr method of endpoint detec-
tion. Chloride and bromide ions are titrated with
Equivalence point standard silver nitrate using chromate ion as indicator,
the endpoint being indicated by the appearance of
brick-red silver chromate.[2]
Volume of AgNO3

Fig. 1 A theoretical precipitation titration curve. Agþ þ Cl
! AgCl ðsÞ

þ 2

Ag þ CrO4 ! Ag2 CrO4 ðsÞ

Endpoint Detection Because silver chromate is more soluble, the Ksp value
(soluble product constant) of silver chromate is not

Technology–
The stoichiometric equivalence point should be

Tooling
exceeded until the precipitation of Cl
is complete.
immediately detectable. This usually requires a large The endpoint can be corrected by using the Mohr
change in some physical or chemical property of the method to standardize the silver nitrate solution
solution. This point in the reaction is often located against pure sodium chloride.
by means of a secondary system, which provides an The Volhard method of endpoint detection involves
observable endpoint. This secondary system must be using Fe3þ ions as the indicator.[3] This procedure
reproducible, clearly identifiable, and ideally coinci- requires a suitably acidic solution to prevent precipi-
dent with the stoichiometric equivalence point. tation of iron(III) as the hydrated oxide. It has the
Because coincidence is not always achieved, the differ- disadvantage that it is useful only for the reaction:
ence between the endpoint and the equivalence point
should be easily measurable. Often, so-called blank solu- SCN
þ Agþ $ AgSCN ðsÞ
tion is used for this correction. A chemical indicator
After the first excess of thiocyanate ions is added, the
indicator reaction is:

Fe3þ þ SCN
$ FeðSCNÞ2þ ðredÞ

pAg

This means that to determine halide ion (except F
)

back titration is required. In this case, a measured
aliquot of standard silver nitrate solution is added to
I− the sample, and the excess silver ion is determined by
back-titration with a standard thiocyanate solution.
An adsorption indicator is typically an organic dye,
such as fluorescein and its derivatives. Most adsorption
indicators are weak acids. Their use is thus confined to
basic, neutral, or slightly acidic solutions in which the
I− + CI− indicator exists predominantly as the anion. Some
cationic adsorption indicators are suitable for titra-
tions in strongly acidic solutions. In this case, adsorp-
tion of the dye and coloration of the precipitate occur
if the precipitate particles possess a negative charge.
Volume of AgNO3 Because the total amount of ions in solution
decreases as the analyte is precipitated, the conduc-
Fig. 2 Titration curve of solution with iodide and chloride. tance of the solution must decrease. For an accurate
 3754 Titrimetry

Applications

Most applications of precipitation titrations are based

on the use of a standard silver nitrate solution and are
therefore sometimes called argentometric methods
(Table 1).
Conductivity

NEUTRALIZATION TITRATIONS

Acid–Base Equilibria

Aqueous solutions always contain hydronium ions

as well as hydroxide ions as a consequence of the
dissociation of water:
End-point and
equivalence point
identical 2H2 O $ H3 Oþ þ OH

Certain solutes cause changes in the concentrations of

Volume of titrant added the two species, often with profound effect on the
Technology–
Tooling

Fig. 3 A conductivity curve for precipitation titration. chemical characteristics of the solution.
Application of the mass law to the dissociation
of water leads to Kw ¼ [H3Oþ][OH
], where Kw is
called the ion-product constant for water. At 25 C
result, the plot of conductivity against volume of
the ion-product constant has the numerical value of
titrant added has the characteristics shown in Fig. 3.
1.0  10
14 mole2/L2. In pure water, the concentra-
This technique is suitable for both dilute and concen-
tions of hydronium and hydroxide ions are identical,
trated solutions as well as for colored solutions.
and pure water is neutral.
The electromotoric force (emf) of a cell depends on
A useful relationship is obtained from the negative
the ionic concentration of the solutions. To locate the
logarithm of both sides of the ion-product constant
equivalence point, the variation in emf is monitored
expression. Thus:
as the concentration of the analyte changes. When
the measured emf is plotted against the total volume
of titrant added, the curve produced is similar to that
log Kw ¼
log½H3 Oþ ½OH

of a titration curve Fig. 4. This technique has all the ¼
log½H3 Oþ 
log½OH

advantages of the conductometric method and gives
an experimental curve from which the endpoint can from which it follows that:
be detected accurately.
pKw ¼
log Kw ¼ pH þ pOH

where pKw represents the negative logarithm of the

ion-product constant of water. At 25 C, pKw is 14;
that is, pH þ pOH ¼ 14.00.
emf

Table 1 Typical applications of precipitation titrations

Analyte Titrant Endpoint detection



þ
Cl , Br , I Ag Potentiometric
Cl
, Br
, I
Agþ Precipitate
I− Br− Cl− Ag þ
SCN Fe(III), potentiometric
Volume of titrant added Cl
, Br
Hg(NO3)2 Precipitate
2

SO4 , MoO42
Pb(NO3)2 Precipitate
Fig. 4 Titration curves for a mixture of halides.
Titrimetry 3755

Buffer Solutions Titration curves of simple systems

A buffer solution resists changes in pH as a result of When both reagent and analyte are strong, the net neu-
dilution or small additions of acids or bases. The most tralization reaction can be expressed as follows:
effective buffer solutions contain high and approxi-
mately equal concentrations of a conjugate acid–base H3 Oþ þ OH
¼ 2H2 O
pair. The resistance of buffer mixtures to pH is chan-
ged by adding acids or bases. The ability of a buffer The hydronium ions in an aqueous solution of a strong
to prevent a significant change in pH is directly related acid derive from two sources: the reaction of the solute
to the total concentration of the buffering species and with water and the dissociation of water itself.
their concentration ratios. The buffer capacity of a Titration of a strong acid with a strong alkali starts
solution is defined as the number of equivalents of with pure acid in the start that is gradually diluted,
strong acid or base needed to cause 1.00 L of the buffer changing increasingly to a neutral salt until only neu-
to undergo a 1.00-unit change in pH.[4] tral salt remains at the equivalence point. Immediately
beyond the equivalence point, the amount of strong
alkali increases. The change in pH near the equivalence
point will be sharp and large. This may or may not be
Acid–Base Indicators at pH 7, depending on the degree of ionization of the
acid and base, that is, their strengths as acid or base.
Endpoint detection in a neutralization titration is ordi- Titration of a weak acid such as acetic acid against
narily based on the abrupt change in pH that occurs strong base gives a titration curve as that shown in

Technology–
near the equivalence point. A non-instrumental method Fig. 5. At first, partly ionized acid is present owing

Tooling
of pH measurement much used in simple titrations to a pH too high for a strong acid. As neutralization
uses indicators. These are generally organic dyes, or continues, more acid ionizes and mainly ionized acid
weak acids or bases, that on dissociation or association and salt are present. Therefore, the pH changes gradu-
undergo internal structural changes at or near the ally; this is called the buffer effect. If a strong acid is
equivalence point of the neutralization, resulting in a titrated with a weak base such as ammonia, the con-
color change. verse occurs.
A list of some compounds possessing acid-base indi- When a weak acid is titrated with a weak base, the
cator properties is shown in Table 2. Ethanol is the titration curve shows a continuously and gradually
common solvent for indicator solutions. changing pH with the addition of base. No region with
a sharp shift in pH is obtained for small additions of
titrant. If a sharp shift occurs, it is still less than 2
Table 2 Some acid–base indicators
pH units and not detectable by indicators. On both
sides of the equivalence point, buffers are present,
Indicator pH range Color change and at the equivalence point, the pH depends on the
Cresol red 0.2–1.8 Red–yellow relative strengths of acid and base.
Thymol blue 1.2–2.8 Red–yellow
8.0–9.6 Yellow–blue
Bromophenol blue 3.0–4.6 Yellow–blue 12
Methyl yellow 2.9–4.0 Red–yellow
Methyl orange 3.1–4.4 Red–orange 10
Congo red 3.0–5.0 Blue–red
Bromocresol green 3.8–5.4 Yellow–blue 8
Methyl red 4.2–6.3 Red–blue
pH

Bromocresol purple 5.2–6.8 Yellow–purple 6

Bromothymol blue 6.0–7.6 Yellow–blue

4
Phenol red 6.8–8.4 Yellow–red
Cresol purple 7.6–9.2 Yellow–purple
2
Thymol blue 8.0–9.6 Yellow–blue
Phenolphthalein 8.3–10.0 Colorless–red
0
Thymolphthalein 9.3–10.5 Colorless–blue Volume alkali added
Alizarin yellow GG 10–12 Colorless–yellow
Fig. 5 A weak acid-strong base titration curve.
 3756 Titrimetry

Titration curves of complex systems methods that involve an acid-base titration as the final
step. Generally, these elements are non-metallic such
Complex systems include solutions containing two as carbon, nitrogen, chlorine, bromine, sulfur, phos-
acids or bases, which contain or consume two or more phorus, and fluorine. However, in addition, similar
protons, and amphiprotic substances that act as both methods exist for several less commonly encountered
acids and bases. A characteristic of all such systems species. In each case, the element is converted to an
is that two or more equilibria must be considered in inorganic acid or base that can be titrated.
describing their behavior. As a consequence, the tech- Numerous inorganic species can be determined by
niques for pH data derivation are often more complex titration with strong acids or bases. For example,
than for simple systems. ammonium salts are determined by conversion to
A titration curve for the titration of weak and strong ammonia with strong base and distillation in the Kjeldahl
acids with strong base is shown in Fig. 6. The stronger apparatus. Also, inorganic nitrates and nitrites can be
acid is neutralized first, along with some of the weaker determined using Kjeldahl method[5] by reducing these
acid. As a result, a curve more sloping is obtained. The species to ammonium ion.
sharp pH change of the strong acid is still obtained, Carboxylic and sulfonic acid groups import acidity to
although the weaker acid interferes. After all the strong organic compounds. Most carboxylic acids and sulfuric
acid has gone (point A), the weaker acid is titrated acids are readily dissolved in water, and their titration
(point B). with a base is straightforward. If solubility in water is
Compounds with two or more acidic or basic func- not sufficient, the acid can be dissolved in ethanol and
tional groups will yield multiple endpoints in a titrated with aqueous base. Aliphatic amines and many
titration, provided the acidic or basic groups differ suf- saturated cyclic amines can be titrated directly with a
Technology–

ficiently in strength. Computational techniques permit solution of a strong acid. Esters are determined by
Tooling

the derivation of reasonably accurate theoretical saponification with a measured quantity of standard
titration curves for polyprotic acids or bases, provided base. The excess base is titrated with standard acid.
the ratio K1/K2 is above 103. K1 and K2 are dis- Many aldehydes and ketones can be assayed with
sociation constants. The titration curve of a dibasic the aid of a solution of hydroxylamine hydrochloride.
weak acid with NaOH resembles that shown in Fig. 6. The liberated hydrochloric acid is titrated with base.
The derivation of a titration curve for a polyfunc- The total salt content of a solution is determined by
tional base is similar to the previous one for acid. converting the salt to an equivalent amount of an acid
Amphiprotic substance, when dissolved in a suitable or a base by passage through a column packed with an
solvent, behaves both as a weak acid and as a weak ion-exchange resin and then titrated with either base or
base. If either its acidic or its basic character predomi- acid, respectively.
nates sufficiently, titration of the species with a strong
base or a strong acid may be feasible.
Applications in Nonaqueous Media

Applications in Aqueous Media Many useful titrations can be performed in glacial

acetic acid, which is used widely for the titration of
Several important elements that occur in organic and aromatic amines, amides, ureas, and other weak nitro-
biological systems are conveniently determined by gen bases. Direct titration of most amino acids with a
standard acid can be performed in glacial acetic acid.
Basic solvents such as ethylenediamine, dimethyl-
12 formamide, pyridine, and dimethylsulfoxide can be
used for acids that are too weak to be titrated in water.
10
Amine salts, inorganic salts, carboxylic acids, phenols,
8 and imides are soluble in ethylenediamine, where they
B
exhibit enhanced acidic characteristics.
pH

6 A Many sulfa drugs such as sulfanilamide, sulfathio-

zole, and sulfathalidine can be titrated in ethylene-
4
amine with tetrabutylammonium hydroxide. The
2 ability of dimethylformamide to dissolve salts, poly-
mers, and many organic compounds accounts for its
0 wide use as a titration solvent. Inert solvents such as
Volume alkali added
acetone can be used for titration of acids, acetonitrile
Fig. 6 Titration of a mixture of a strong and a weak acid for both acids and bases, and ethyl acetate for
with alkali. amines. A suitable non-aqueous medium such as methyl
Titrimetry 3757

isobutyl ketone makes it possible to discriminate and cerium(III) can be calculated from the amount
among various mineral acids that are not leveled by of titrant added.
the solvent as they are in water. Applying the Nernst equation for the iron(III)/
iron(II) couple gives a value for the potential of
the system directly. Using the couple cerium(IV)/
OXIDATION–REDUCTION TITRATIONS
cerium(III) would give the same answer, but it would
first be necessary to calculate a value for the concen-
Equilibrium
tration of cerium(IV), which in turn would require
evaluation of the equilibrium constant for the reaction.
Oxidation of a substance (element or a compound)
As shown in Fig. 7 there is a rapid change in the
involves the loss of electrons. Conversely, reduction
value of E as the titration is proceeded through the
of a substance involves the gain of electrons. Thus, in
endpoint. In fact, the titration curve has the same gen-
oxidation–reduction system, electron transfer occurs.
eral form as that of an acid–base titration. An exact
In titration of iron(II) with cerium(IV):
value for the endpoint can be calculated using the
Nernst equations for the half-reactions.
Ce4þ þ Fe2þ $ Ce3þ þ Fe3þ The shape of the curve for an oxidation–reduction
titration depends on the nature of the system under
equilibrium is attained after each addition of titrant. consideration. The titration curve in Fig. 7 is symmetric
After the first addition of cerium(IV), all four species about the equivalence point because the molar ratio of
will be present in the solution in amounts dictated by oxidant to reductant is equal to unity. An asymmetri-
the equilibrium constant of the reaction. At chemical cal curve results if the ratio differs from this value.
equilibrium of an oxidation–reduction system, the

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Solutions containing two oxidizing or reducing agents

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electrode potentials E for the two half-reactions are yield titration curves containing two inflection points if
identica. Thus, at any point in the titration: the standard potentials for the two species are different
by more than approximately 0.2 V. Fig. 8 shows
ECe4þ ¼ EFe3þ ¼ Esystem the titration curve for a mixture of iron(II) and
titanium(III) with cerium(IV). The first additions of
If the solution contains a reversible oxidation– cerium are used by more readily oxidized titanium(III)
reduction indicator as well, its potential must be the ion, thus, the first step in the titration curve corre-
same as that for the system: sponds to titanium and the second to iron.
EIn ¼ ECe4þ ¼ EFe3þ ¼ Esystem
Oxidation–Reduction Indicators
Titration Curves Specific indicators owe their behavior to a reaction
with one of the participants in the titration. The best-
The potential of a system can be measured experimen- known specific indicator is starch, which forms a dark
tally by determining the emf of a suitable cell. Thus, for blue complex with triiodide ion. Also, potassium
the titration of iron(II) with cerium(IV), the cell is: thiocyanate is used as a specific indicator, for example,
 in titration of iron(III) with solutions of titanium(III)
SCEjCe4þ ; Ce3þ ; Fe3þ ; Fe2þ jPt sulfate.

The potential of the platinum electrode versus a satu-

rated calomel electrode (SCE) is determined by the
affinity of Fe3þ for electrons:

Fe3þ þ e ! Fe2þ
E (V)

and that of Ce4þ:

Ce4þ þ e ! Ce3þ

In deriving Esystem data for a titration curve,

either ECe4þ or EFe3þ can be used. Short of the equiv- Volume titrant added
alence point, the concentration of cerium(IV) is vanish-
ingly small. The concentrations of iron(II), iron(III), Fig. 7 Titration curve for iron(II) with cerium(IV).
 3758 Titrimetry

commonly used with solutions that are 0.1 N or greater

in mineral acid. Under those conditions, the product is
manganese(II):

MnO4
þ 8Hþ þ 5e $ Mn2þ þ 4H2 O

However, the tendency of permanganate to oxidize

E (V)

chloride ion is a disadvantage because hydrochloric

acid is such a useful solvent and, furthermore, KMnO4
solutions have limited stability.
For example, determinations of Sn, H2O2, Fe, V,
Mo, W, U, Ti, Mg, Ca, Zn, Co, Pb, Ag, K, Na, and HNO2
potassium permanganate are used as oxidizing agents.
A sulfuric acid solution of cerium(IV) is nearly as
potent an oxidizing reagent as is permanganate and
can be substituted for the latter in most of the appli-
cations described with potassium permanganate.
Volume titrant added Cerium(IV) does not oxidize chloride ion at a detect-
able rate as does permanganate, and the reagent is
Fig. 8 Titration of iron(II) and titanium(III) with cerium(IV). indefinitely stable. On the other hand, the color of
Technology–

cerium(IV) is not sufficiently intense to serve as an

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Oxidation–reduction indicators indicator. In addition, the reagent cannot be used in

neutral or basic solutions.
A redox indicator is essentially a compound that has Use of potassium dichromate is limited because of
oxidized and reduced forms with different, and pre- its lesser oxidizing strength and slowness of some of
ferably intense, colors (Table 3). The half-reaction its reactions. This technique is used for determination
responsible for color change in a typical oxidation– of iron(II), nitrate, chlorate, permanganate, and
reduction indicator can be written as: organic peroxides, among others.
Iodine, a relatively weak oxidant, is used for the
selective determination of strong reducing agents.
IndicatorðoxidizedÞ þ ne ¼ IndicatorðreducedÞ Iodine can be used for determination of As, Sb, Sn,
H2S2, SO2, S2O32
, and N2H4, among others. Its great
advantage is its ready availability as a sensitive and
Applications reversible indicator. Its disadvantages include the low
stability of iodine solutions and the incompleteness
Standard oxidants of reactions between iodine and many reducing agents.
Potassium bromate is a convenient source of bro-
The powerful oxidant potassium permanganate, mine in organic analysis. Few organic compounds
KMnO4, is a widely used oxidizing agent. It is most react sufficiently rapidly for a direct titration, and,

Table 3 Some redox indicators

Indicator Color change oxidized–reduced Transition potential, V
Indigomonosulphonate Blue—colorless þ0.26
Phenosufranine Red—colorless þ0.28
Indigotetrasulphonate Blue—colorless þ0.36
Methylene blue Blue/green—colorless þ0.53
Diphenylamine Violet—colorless þ0.76
p-Ethoxychrysoidine Yellow—red þ0.76
Diphenylamine sulfonic acid Red/purple—colorless þ0.85
Eriocaucin A Red—yellow/green þ0.98
1,10-Phenanthroline iron(II) Pale blue—red þ1.11
2,3-Diphenylamine dicarboxylic acid Blue/violet—colorless þ1.12
Titrimetry 3759

thus, a measured excess of the bromate is added to the (or stability) constants (Fig. 9). Equilibrium between a
sample, and after the reaction is complete, the excess metal ion M, possessing a coordination number of 4,
bromine is back-titrated with an arsene(III) solution. and the quadridentate ligand D can be represented by:
Some organic compound analyzed using bromine
substitution are, for example, phenol, p-chlorophenol, M þ D $ MD
salicylic acid, acetylsalicylic acid, m-cresol, aniline,
sulfanilic acid, and b-naphtol. Bromine addition is used
In similar manner, the equilibrium between M and
most often in the estimation of olefinic unsaturation in
bidentate B can be represented by:
fats, oils, and petroleum products.
M þ 2B $ MB2
Reductants

Because of the readiness of reducing agents to react Overall formation of MB2 occurs in two steps and
with atmospheric oxygen, the titrations must be carried involves the intermediate formation of MB:
out in and the reagents must be stored under an inert
atmosphere. Alternatively, a stable standard oxidizing M þ B $ MB
agent can be used for titration of an aliquot of the MB þ B $ MB2
reductant to determine the current concentration of
the reducing agent.
A variety of oxidizing agents such as Cr(VI), Ce(IV), The formation constants for these individual processes
Mo(VI), NO3
, NH2OH, and organic peroxides can are:

Technology–
be determined by reaction with a measured excess of

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standard iron(II) solution. Standard potassium dichro- K1 ¼ ½MB=½M½B
mate is frequently used for the back-titration.
Iodide ion is a moderately effective reducing agent. and
In its applications, a standard solution of sodium thio-
sulfate is used to titrate the iodine liberated by reaction K2 ¼ ½MB2 =½MB½B
of the analyte with an unmeasured excess of potassium
iodide. Some substances determined by using iodo-
metric method are IO4
, IO3
, BrO3
, ClO3
, Br2,
Cl2, O2, O3, Cu2þ, NO2
, and organic peroxide.
The water content in solids and many organic acids,
alcohols, esters, ethers, anhydrides, and halides can be 1:1
determined using Karl–Fischer reagent,[6] composed of
iodine, sulfurdioxide, pyridine, and methanol. 2:1

COMPLEX-FORMATION TITRATIONS 4:1

pM

Metal ions can act as electron-pair acceptors, reacting

with electron donors to form coordination compounds
or complex ions. The donor species, or ligand, must
have at least one pair of unshared electrons to form C 4:1
the bond. Remarkable growth in the analytical appli-
cations of complex-formation reactions is attributable
B
to a particular class of coordination compounds called 2:1
chelates. These compounds are made by the reaction of
a metal ion and a ligand that contains two or more
donor groups. The properties of chelates can differ A
markedly from the parent cation. 1:1

Reagent volume
Titration Curves
Fig. 9 Curves for complex formation titrations. Titration of
The data necessary to plot theoretical p-functions the tetradentate ligand D (curve A), bidentate ligand (curve
versus reagent volumes require the use of formation B), and unidentate ligand (curve C).
 3760 Titrimetry

The equilibrium constant for the overall reaction is Numerous tertiary amines that also contain car-
given by the product of the individual steps: boxylic acid groups form remarkably stable chelates
with many metal ions. Ethylenediamine tetra-acetic
Koverall ¼ ½MB2 =½M½B2 ¼ K1 K2 acid (EDTA) can be used for determination of 40
elements by direct titration using metal-ion indicators
The complex between M and a simple, unidentate for endpoint detection.[8] Direct titration procedures
ligand A results in the overall equilibrium: are limited to metal ions that react rapidly with EDTA.
Back titration procedures are useful for the analysis of
M þ 4A $ MA4 cations that form very stable EDTA complexes and for
which a satisfactory indicator is not available. EDTA
This process occurs also in a stepwise manner, and the
is also used for determining water hardness; the total
equilibrium constant for the overall reaction is there-
concentration of calcium and magnesium expressed
fore numerically equal to the product of the constants
in terms of the calcium carbonate equivalent.
for the four constituent processes.

Endpoint Detection POTENTIOMETRIC TITRATIONS

In some complex-formation titrations, the endpoint is In potentiometric titrations, the activity of one species
noted by the formation or disappearance of a solid is continuously monitored as it changes during the
phase. For example, in the titration of cyanide with sil- course of the titration, for example, in the titration:
ver ion, the solution remains clear, but the first excess
Technology–

of silver causes formation of a white solid that marks AgNO3 þ NaCl $ AgCl þ NaNO3
Tooling

the endpoint. The electron-donor groups of most com-

mon ligands tend to combine not only with metallic where silver nitrate is added to aqueous sodium chlor-
ions but also with protons; thus, the equivalence point ide. The change in activity of the Cl
can be moni-
in a complex-formation titration is often accompanied tored. The potential between the reference electrode
by a marked change in pH, which can be detected with and the indicator electrode is usually measured at the
an acid–base indicator. start of and after the addition of small amounts of
Formation or disappearance of a soluble colored titrant. A reproducible equilibrium is of little concern.
complex can also indicate an endpoint. Many reagents Requirements for reference electrodes are greatly
that form colored complexes with certain metals have relaxed. In potentiometric titrations, accuracy is higher
been developed only for use as indicators in these titra- than that in direct potentiometric measurements
tions. If the cation being titrated produces a color with because measured potentials are used to detect rapid
the indicator, the endpoint will be characterized by the changes in activity that occur at the equivalence point
disappearance of this color. When the cation does not of the titration. The change in emf versu titration
give a colored complex, a second cation that does is volume is more interesting than the absolute value of
introduced, and the first excess of titrant then deco- the emf. In potentiometric titrations, the influence of
lorizes this complex. liquid-junction potentials and activity coefficients is
minimized. The acid and alkali errors of the glass elec-
trode occur at extremes of pH and have little effect
Some Applications near the interesting equivalence point.

Inorganic complexing reagents such as Hg(NO3)2,

AgNO3, NiSO4 and KCN can be used for complex- Titration Curves and Endpoint Detection
formation titrations. Mercury(II) ion forms neutral
complexes with most of the anions that precipitate In potentiometric titrations, the titration curve can be
with silver nitrate such as Br
, Cl
, SCN
, CN
and followed point by point, plotting as the ordinate suc-
thiourea. cessive values of the cell emf versus the corresponding
AgNO3 reacts with CN
forming Ag(CN)2
. Iodine volume of titrant added as the abscissa (Fig. 10). The
used as indicator and endpoint is detected by forma- titrant should be added in the smallest accurately
tion of solid AgI. NiSO4 can also be used for determi- measurable increments that provide an adequate den-
nation of CN
with AgI as indicator. The endpoint is sity of points, particularly in the vicinity of the equiv-
also detected here by formation of AgI. KCN reacts alence point. The greatest change in emf occurs around
with Cu2þ, Hg2þ, and Ni2þ forming Cu(CN)4, the equivalence point. The most straightforward
Hg(CN)2 and Ni(CN)4. Various indicators for end- method takes the midpoint in the steeply rising portion
point detection can be used.[7] of the curve as the endpoint.
Titrimetry 3761

Antilog (E/S)
Potential E (mV)

O
Equivalence point
Volume of addition

Fig. 11 Gran’s plot for a potentiometric titration.

Volume of addition
The first antilog term is a constant and therefore:
Fig. 10 Potentiometric titration curves.
Ci ¼ const  ½antilogðE=SÞ

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Tooling
A second approach is to calculate the change If the species i is disappearing during the course of the
in potential-per-unit change in volume in reagent titration, then it will do it linearly. The antilog term
(DE/DV). By inspection, the endpoint can be located reduces linearly, and a plot of antilog (E/S) against
from the inflection point of the titration curve. This volume of reagent added should give a straight line
is the point that corresponds to the maximum rate of whose intercept with the volume axis will be the equiv-
change of cell emf per unit volume of titrant added alence point, as shown in Fig. 11.
(usually 0.05 or 0.1 mL). The first-derivative method
is based on the sigmoid shaped curve. Applications
The second-derivative method is an extension of the
first-derivative method. The second-derivative of the Precipitation and complex-formation titrations
data changes sign at the point of inflection in
the titration curve. This change is often used as the The indicator electrode for a precipitation titration is
analytical signal in automatic titrators. often the metal from which the reacting cation is
Grans method[9,10] involves the use of the Nernst derived. Membrane electrodes that are sensitive to
equation: one of the ions involved in the titration process may
be used. For example, fluoride-sensitive membrane
E ¼ E þ S log Ci electrode is used in the determination of the fluoride
content of toothpastes. Lanthanum(III) solution is
where used as a precipitant.
Silver nitrate is the most commonly used reagent for
S is electrode slope; E is E0  ð0:0591=nÞ log i; and precipitation titration, including potentiometric deter-
mination of endpoint. Argentometric methods exist
and E0 is the constant incorporating the potential of for the determination of halides, halogenoids, mercap-
the reference electrode and the standard potential of tans, sulfides, phosphates, oxalates, and arsenates. A
the half-cell containing the solution under investi- silver electrode serves as indicator for the potentio-
gation and the ion selective electrode. The rearrange- metric determination of these ions. If the concentration
ment of this equation gives: of reagent and analyte is 0.1 M or higher, then a calo-
mel electrode can be used. In dilute solutions, the slight
log Ci ¼ ðE
E Þ=S leakage of chloride ions from the salt bridge could be a
source of significant error, but not in more concen-
or trated solutions.
Mercury electrode can be used for the potentio-
Ci ¼ antilog½ðE
E Þ=S metric determination of 29 divalent, trivalent, and
¼ ½antilogð
E =SÞ½antilogðE=SÞ tetravalent cations with EDTA.[11,12]
 3762 Titrimetry

Neutralization titrations Another method uses a preset equivalence point

potential applied across the electrodes by means of a
Potentiometric acid–base titrations are particularly calibrated potentiometer. A difference between this
useful for the analysis of mixtures of acids or poly- potential and that of the electrodes causes an ‘‘error’’
protic acids (or bases) because often, discrimination signal, which is amplified. This causes the electronic
between the endpoints can be made. An approximate switch to close, permitting a flow of electricity through
numerical value for the dissociation constant of a weak the solenoid-operated valve of the burette. As the
acid or base can be estimated from potentiometric signal approaches zero, the flow of titrant ceases as
titration curves. In theory, this quantity can be the current to the solenoid is switched off.
obtained from any point along the curve, but it is Second-derivative titrators have the advantage that
most easily derived from the pH at the point of half- no preknowledge of the equivalence point potential is
neutralization. required. The signal processor calculates the second
In the case of weak acid HA, it can be assumed that derivative of the electrode potential of the indicator
at the midpoint, [HA] ¼ [A
], and thus: electrode. Change in the sign of the second derivative
causes a switching device to turn off the flow of the
Ka ¼ ½Hþ ½A
=½HA ¼ ½Hþ  titrant.
A fully automated unit accepts a series of samples
which gives placed on a turntable. After each titration, the turn-
table rotates, indexes the next sample beneath the elec-
pKa ¼ pH trode assembly, and starts the titration.
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It must be noted that this constant is not correct

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because activities should be used in calculations:

Ka ¼ ½A
fA =½HAfHA CONDUCTOMETRIC TITRATIONS

and because Theory

½HA and ½A
 are equal Ka ¼ fA =fHA Conduction of an electric current through an electro-
lyte solution involves migration of positively charged
species toward the cathode and negatively charged
Oxidation–reduction titrations species toward the anode. The conductance of a solu-
tion is ameasure of the current flow that results with
In oxidation–reduction titrations, an electrode poten- application of a given electrical force. It is directly
tial related to the concentration ratio between the oxi- dependent on the number of charged species the solu-
dized and reduced forms of either of the reactants is tion contains.[14] The conductance L of a solution is
determined as a function of the titrant volume. The also the reciprocal of the electrical resistance and has
indicator electrode must be responsive to at least one the units of ohm:
of the couples involved in the reaction. Indicator elec-
trodes for oxidation–reduction titrations are generally L ¼ 1=R
constructed from platinum, gold, mercury, or pal-
ladium. The metal chosen must be unreactive with
respect to the components of the reaction. The indi- where R is the resistance in ohms.
cator metal is merely a medium for electron transfer. The conductance is directly proportional to the
length (l) of a uniform conductor. Thus:

Automatic Titrators L ¼ kA=l

An entire titration can be performed automatically by
titrators equipped with microcomputers and analog- where k is a proportionality constant called the specific
to-digital converters and using dedicated software.[13] conductance, and A is the surface area.
The most widely used apparatus for automatic reagent The equivalent conductance, L, is defined as the
addition consists of a calibrated syringe that is conductance of a 1-g equivalent of solute contained
activated by a motor-driven micrometer screw. The between electrodes spaced 1 cm apart. It is equal to L
volume is determined from the number of turns the when 1-g equivalent of solute is contained between
screw makes during the titration. electrodes spaced 1 cm apart. The volume V of solution
Titrimetry 3763

(cm3) that will contain 1 gram equivalent solute is given Applications

by:
Acid–base titrations
V ¼ 1000=C
Neutralization titrations are particularly well-adapted
to the conductometric titration because of the very
where C is the concentration in equivalents per liter. high conductance of the hydronium and hydroxide
Volume can also be expressed in terms of the ions compared with the conductance of the reaction
dimensions of the cell: products. In neutralization of strong acids, hydronium
ions are being replaced by an equivalent number of less
V ¼ lA mobile sodium ions, and the conductance decreases as
a result of this substitution. At the equivalence point,
the concentration of hydronium and hydroxide ions
when l is 1 cm
are at a minimum, and the solution exhibits its lowest
conductance. After the endpoint, a reversal of slope
V ¼ A ¼ 1000=C occurs as the sodium ion and the hydroxide ion con-
centration from the excess base increase. There is an
when L ¼ kA=1 excellent linearity between conductance and the
and L ¼ L; then L ¼ 1000k=C volume base added, except at very near equivalence
point region. Very dilute solutions can be analyzed
accurately.
A conductometric titration involves measurement of

Technology–
In conductometric titrations, a change in conduc-

Tooling
the conductance of the sample after successive addi-
tance is caused after the endpoint owing to the increase
tions of reagent. The endpoint is determined from a
in concentration of the mobile hydroxide ion. Very
plot of either the conductance or the specific conduc-
weak acids (such as boric acid) and moderately weak
tance as a function of the volume of added titrant.
acids (such as acetic acid) can be titrated using conduc-
Throughout a titration, the volume of the solution is
tometric titration.
always increasing. Unless the conductance is corrected
If the titrant is a weak electrolyte (such as
for this effect, non-linear titration curves result. The
ammonia), the curve is essentially horizontal past the
titrant should be at least 10 times as concentrated as
equivalence point, which causes less uncertainty to
the solution being titrated to keep the volume change
the extrapolation of a curve. In titration of a weak
small. Some temperature control is ordinarily required
base, such as acetate ion, with a strong acid, a salt
during a conductometric titration because the tempera-
and undissociated acetic acid are formed. After the
ture coefficient for conductance measurements is
endpoint is passed, a sharp rise in conductance attends
approximately 2% per  C.
the addition of excess hydronium ions. Salts whose
acidic or basic character is too weak to give satisfac-
tory endpoints with indicator are conveniently titrated
Titration Curves with the conductometric method. The conductometric
titration of a mixture of two acids that differ in degree
Titration curves for conductometric titrations take a of dissociation is frequently more accurate than a
variety of shapes, depending on the chemical system potentiometric titration.
under investigation. In general, they are characterized
by straight line portions with dissimilar slopes on
either side of the equivalence point, as shown pre- Precipitation and complex-formation
viously in Fig. 3. To establish a conductometric end- titrations
point, after correcting for volume changes, the
conductance data are plotted as a function of titrant Fig. 12 illustrates the titration of sodium chloride with
volume. The two linear portions are then extrapolated, silver nitrate. After all chloride is precipitated, the
and the point of intersection is taken as the equivalence addition of excess silver nitrate causes a rapid increase
point. Frequently, reactions fail to proceed to absolute in conductivity. The slope of the initial portion of the
completion, and the conductometric titration curves curve may be either downward or upward, depending
invariably show departures from strict linearity in the on the relative conductance of the ion being determ-
region of the equivalence point. ined and the ion of like charge in the reagent that
An advantage of the conductometric titration is replaces it. Slow reactions and coprecipitation are
that it can be used for titrations based on relatively sources of difficulty with precipitation and complex-
unfavorable equilibria. formation titrations.
 3764 Titrimetry

The endpoint may be detected by addition of

colored indicators, provided the indicator itself is not
electroactive. Potentiometric and spectrophotometric
indication is used in acid–base and oxidation–
reduction titrations. Amperometric procedures are
applicable to oxidation–reduction and ion-combination
Conductance

reactions especially for dilute solutions.

The most obvious advantage of coulometric
titration is the elimination of problems associated with
the preparation, standardization, and storage of
standard solutions. This is particularly important for
labile reagent substance such as chlorine or bromine.
Another advantage is the requirement of a small
quantity of reagent. Instrumentation is simple; a single
constant-current source can be used to generate pre-
cipitation, oxidation–reduction, or neutralization
Volume of AgNO3 reagents. Furthermore, coulometric titration is easily
automated.
Fig. 12 Titration of chloride ion with silver nitrate. Typical sources of error in coulometric titrations are
variation in the current during electrolysis and errors
COULOMETRIC TITRATIONS in measurement of current and time. Departure of
Technology–

the process from 100% current efficiency is the primary

Tooling

Theory error source. In some cases, the equivalence point is

not the endpoint of the titration.
A coulometric titration uses an electrolytically gener-
ated titrant for reaction with the analyte. In some
analyses, the active electrode process involves only Applications
generation of the reagent. In other titrations, the
analyte may also be directly involved at the generator Neutralization titrations
electrode.[15] The current in a coulometric titration
is carefully maintained at a constant and accurately Both weak and strong acids can be titrated with a high
known level. The product of this current and the time degree of accuracy using electrogenerated hydroxide
required to reach the equivalence point for the reaction ions. In this application, the platinum anode must be
yield the number of coulombs and thus the number of isolated by some sort of diaphragm to eliminate poten-
equivalents involved in the electrolysis. The coulomb tial interference from the hydrogen ions produced.
(C) is the quantity of electricity that is transported in Alternatively, chloride or bromide ions can be added
1 by a constant current of 1 ampere. The Faraday con- to the solution, and a silver wire used as an anode.
stant (F) is the quantity of electricity that produces one The reaction at this electrode is:
equivalent of chemical change at an electrode.[16]
The so-called primary titration technique is Agþ þ Br
$ AgBr þ e
attempted only with electrodes of silver metal, silver–
silver halide, or mercury amalgams, which are the Thus, this anode product does not interfere with the
source of the electrogenerated species. The substance neutralization reaction.
to be determined reacts directly at the electrode or with Strong and weak bases can be titrated with hydro-
a reactant electrogenerated from the working elec- gen ions generated at a platinum anode.
trode. This class of titrations is limited generally to Here also the cathode must be isolated from the sol-
non-diffusible reactants such as mercury amalgams, ution to prevent interference from the hydroxide ions
silver ions generated by anodization of silver metal, produced at that electrode.
and halides liberated by reduction of the appropriate
silver–silver halide electrode. Precipitation and complex-formation titrations
In secondary coulometric titrations, an oxidation–
reduction buffer serves as the titrant precursor. An Anodically generated silver ions can be used for pre-
active intermediate from the titrant precursor must cipitation titrations of various substances. A cell con-
first be generated with 100% efficiency by the electrode structed from a piece of heavy silver wire can be
process. The intermediate must react rapidly and used. Substances precipitated include Cl
, Br
, I
,
completely with the substance being analyzed. and mercaptans. Similar applications using mercury(I)
Titrimetry 3765

Table 4 Coulorimetric titrations of oxidation–reduction is added, and the titrant is again added until the speci-
reactions fied current level is reached.
Reagent Substance determined A correction for dilution is necessary to attain a lin-
Br2 As , Sb3þ, U5þ, NH3, N2H4,
3þ ear relationship between current and volume of titrant.
NH2OH, phenol, aniline By working with a reagent that is 10-fold more concen-
trated than the solution being titrated, this correction
Cl2 As3þ, I

becomes negligible. Amperometric and conductometric
I2 As3þ, Sb3þ, S2O32
, H2S titrations are similar in the respect that the data for
Ce 4þ
Fe2þ, Ti3þ, U4þ, As3þ, I
, Fe(CN)63
each are collected well away from the equivalence
Fe2þ Cr6þ, Mn7þ, V3þ, Ce4þ point. Therefore, reactions that are relatively incom-
Ti 3þ
Fe3þ, V5þ, Ce4þ, U6þ plete can be used.
C2Cl32
V5þ, Cr6þ, IO3

Titration Curves

Typical forms of amperometric titration curves are

ion formed at a mercury anode have been used for the shown in Fig. 13. A titration in which the substance
determination of Cl
, Br
, and I
. being analyzed reacts (is reduced) at the electrode while
The complexing ability of ethylenediaminetetra- the reagent does not is shown in Fig. 13A. A sufficiently
acetic acid (EDTA, H4Y) has been exploited in the high potential is applied to give a diffusion current for
coulometric titration of metal ions. The method the substance, and a linear decrease in current is
depends on the reduction of the mercury(II) or cad-

Technology–
observed as substance ions are removed from the solu-
mium chelate of EDTA and on the titration of the

Tooling
tion by precipitation. The curvature near the equi-
metal ion (for example, magnesium) to be determined valence point reflects the incompleteness of the
by the anion of EDTA that is released. analytical reaction in this region. The endpoint is
obtained by extrapolation of the linear portions. Nearly
Oxidation–reduction titrations a mirror image curve is obtained for a titration in which
the reagent reacts (is reduced) at the microelectrode, and
Applications of coulometric titrations involving the substance being analyzed does not (Fig. 13B). The
oxidation–reduction reactions are shown in Table 4. third common curvature for amperometric titrations is
Electrogenerated oxidizing agents such as bromine obtained when both reagent and subtance analyzed react
have proved to be useful, especially in organic analysis. (are reduced) at the electrode Fig. 13C.

Amperometric Titrations with Two

AMPEROMETRIC TITRATIONS Polarized Microelectrodes
Theory A modification of amperometric method involves the
use of two stationary microelectrodes immersed in a
Polarographic methods can be used to estimate the well-stirred solution of a sample.[17] A small potential
equivalence point of a reaction, provided that at least is applied between these electrodes, and such current
one of the participants or products of the titration is that flows is followed as a function of the volume of
oxidized or reduced at the microelectrode. When the reagent added.
potential applied across the two electrodes is main-
tained at some constant value, the current may be
measured and plotted against the volume of the titrant, A B C
thus, the term amperometric titration. In the case of
working electrode-reference electrode pair, the poten-
Current

Current

Current

tial of the indicator electrode is maintained at a

constant value with respect to a reference electrode,
measuring a limiting current, which is proportional
to the concentration of one or more of the reactants
or products of the titration.
Volume of titrant added
Amperometric titration is easily automated. The
titrator can be programmed to shut off when a speci- Fig. 13 Amperometric titration curves: (A) analyte is
fied current level is reached. Titrant is run into a blank reduced, reagent is not; (B) reagent is reduced, analyte is
solution until a specified current is reached, the sample not; (C) both reagent and analyte are reduced.
 3766 Titrimetry

Oxidation–reduction titrations Table 5 Applications of amperometric titrations with

precipitation products
Twin polarized platinum microelectrodes are con- Reagent Substance determined
veniently used for endpoint detection for oxidation– AgNO3 Cl
, Br
, I
, CN
, RSH
reduction titrations. Consider a titration curve for
Cupferron Cu2þ, Fe3þ
oxidation–reduction titration where both reactants
behave reversibly at the electrodes. An example of this Dimethylglyoxime Ni2þ
kind of titration is titration of iron(II) with cerium(IV) Pb(NO3)2 SO42þ, MoO42þ, F
, Cl

(Fig. 14A). At the starting point of the titration, no 8-Hydroxyquinoline Mg2þ, Cu2þ, Zn2þ, Cd2þ,
current is observed because no suitable cathode reac- Al3þ, Bi3þ, Fe3þ,
tant is available. With addition of cerium(IV), a K2CrO4 Pb2þ, Ba2þ
mixture of iron(II) and iron(III) is produced, which
permits the passage of current. Beyond the midpoint
in the titration, iron(III) becomes in excess, and the precipitation reagent (Table 5). Short of the equival-
current is then regulated by decreasing iron(II) con- ence point, essentially no current exists when a small
centration. At the equivalence point, the current potential is applied between two such electrodes during
approaches zero because iron(III) are present, and a titration of substance analyzed with silver ions
the applied potential is not great enough to cause these because no easily reduced species is present in the solu-
to react at the electrode. Beyond the equivalence point, tion. After equivalence, the cathode becomes depolar-
the current rises again because both cerium(III) and ized owing to the presence of a significant amount of
cerium(IV) are present to react at the electrodes. silver ions that can react to give silver. Current is
Technology–

In cases where only reagent behaves reversibly, a permitted as a result of this half-reaction and the
Tooling

different form for titration curve is obtained. Although corresponding oxidation of silver at the anode. The
the reagent added can serve as an anode reactant, magnitude of the current is directly proportional to
no cathode reactant is available because of the slow the concentration of the excess reagent.
rate at which the substance analyzed is reduced at a
platinum surface. Therefore, no current is observed.
Beyond the equivalence point, depolarization of the
cell can occur, and the current is dependent on the con- PHOTOMETRIC TITRATIONS
centration of the reagent (Fig. 14B). In cases where
only the species titrated behave reversibly, before The change in absorbance of a solution may be used
equivalence point, a current is observed that depends to follow the change in concentration of a radiation-
on the concentration of the species present in lesser absorbing constituent during a titration.[18] The absor-
amount. At equivalence point, a zero current is bance is directly proportional to the concentration
reached, and beyond the equivalence point, no current of the absorbing constituents. A plot of absorbance
is observed because the reagent does not behave rever- versus titrant consists, if the reaction is complete, of
sibly at the electrodes (Fig. 14C). two straight lines that intersect at the endpoint. If the
reaction is appreciably incomplete, extrapolation of the
Precipitation titrations two linear segments of the titration curve establishes
the intersection and endpoint volume.
Twin silver microelectrodes permit observation of the Photometric titrations have several advantages over
endpoint for various titrations using silver nitrate as a direct photometric determination. The presence of
other absorbing species at the analytical wavelength
does not necessarily cause interference because only
the change in absorbance is significant. However, if
the absorbance of non-titratable components is intense,
A B C the absorbance readings are shifted to the undesirable
upper end of the absorbance scale. Ideally, only a
Current
Current

Current

End End
point End point single absorber is present among the reactant, titrant,
point
and products.
The analytical wavelength is selected to avoid the
Volume of titrant added
interference caused by other absorbing substances.
Fig. 14 Amperometric titration curves with twin polarized Also, there is a need for a molar absorbtivity that
electrodes: (A) both reactants behave reversibly at the elec- causes the change in absorbance during the titration
trode; (B) only reagent behaves reversibly; (C) only analyte to fall within a convenient range. Often, the chosen
titrated behaves reversibly. wavelength lies well apart from an absorption maximum.
Titrimetry 3767

A B measurement can be changed easily by simply chan-

ging the wavelength or the length of the cell path, a

Absorbance
Absorbance
feature that also makes photometric titration attrac-
tive. Indicators can be used in titrations where self-
indicating systems are lacking.
End-point

C D REFERENCES

1. Grant, J. Sutton’s Volymetric Analysis, 13th Ed.; Butter-

Absorbance

Absorbance
worth’s Scientific Publications: London, 1955; 28–29.
2. Skoog, D.A.; West, D.M. Fundamentals of Analytical
Chemistry, 2nd Ed.; Holt, Rinehart, Wilston: London,
1972; 225.
3. Skoog, D.A.; West, D.M. Fundamentals of Analytical
Chemistry, 2nd Ed.; Holt, Rinehart, Winston: London,
Volume titrant added 1972; 226–227.
4. Laitinen, H.A. Chemical Analysis; McGraw-Hill: New
York, 1960; 37.
Fig. 15 Different shapes of photometric titration curves. 5. Grant, J. Sutton’s Volymetric Analysis, 13th Ed.; Butter-
worth’s Scientific Pulications: London, 1955; 140.
6. Mitchell, J. Anal. Chem. 1951, 23, 1069
Titration Curves 7. Meites, L. Handbook of Analytical Chemistry; McGraw-
Hill: New York, 1963; 3–226.
8. Reilley, C.N.; Barnard, A.J., Jr. Handbook of Analytical
Volume change, caused by addition of titrant, is sel-

Technology–
Chemistry; McGraw-Hill: New York, 1963; 3–166 to 3–200.

Tooling
dom negligible, and straight lines for titration curve 9. Gran, G. Acta Chem. Scand. 1950, 4, 559.
are obtained only if there is correction for dilution. 10. Gran, G. Analyst 1952, 77, 661.
11. Reilley, C.N.; Schmid, R.W. Anal. Chem. 1958, 30, 947.
If correction is not made, the lines are curved down- 12. Reilley, C.N.; Schmid, R.W.; Lamson, D.W. Anal. Chem.
ward toward the volume axis, and erroneous inter- 1958, 30, 953.
sections are obtained. Use of a microsyringe and a 13. Foreman, J.K.; Stockwell, P. Automatic Chemical Analysis;
Wiley: New York, 1975; 42–62.
relatively concentrated titrant is desirable. 14. Shadlovsky, T.; Weisserger, A. Physical Methods of
Possible shapes of photometric titration curves are Organic Chemistry, 3rd Ed.; Interscience: New York,
shown in Fig. 15. Curve A is typical of the titration 1960; 1, 3011–3048.
15. Clem, R.G. Ind. Research 1973, (September), 50.
where the titrant alone absorbs and where the absor- 16. Skoog, D.A.; West, D.M. Fundamentals of Analytical
bance readings are taken at the wavelength of the Chemistry, 4th Ed.; Holt, Rinehart, Winston: Hong Kong,
titrant. Curve B is characteristic of systems where the 1986; 443–444.
17. Lingane, J.J. Electroanalytical Chemistry, 2nd Ed.; Inter-
product of the reaction absorbs, and curve C is typical science: New York, 1958; 280–294.
for reactions where the analyte is converted to a 18. Goddu, R.F.; Hume, D.N. Anal. Chem. 1954, 26, 1679.
non-absorbing product. When a colored analyte is
converted to a colorless product by a colored titrant,
curves similar to D are obtained. BIBLIOGRAPHY

Headridge, J.B. Photometric Titrations; Pergamon Press:

Applications New York, 1961.
Laitinen, H.A.; Harris, W.E. Chemical Analysis, 2nd Ed.;
McGraw-Hill: New York, 1975.
Areas of particular applicability are for solutions so Lingane, J.J. Electroanalytical Chemistry, 2nd Ed.; Interscience:
dilute that the indicator blank is excessive or when New York, 1958.
the color change is not sharp perhaps because titration Ma, T.S.; Hassan, S.S.M. Organic Analysis Using Ion-Selective
Electrodes; Academic Press: London, 1982; 1, 2.
reactions that are incomplete in the vicinity of the Rechnitz, G.A. Bioanalysis with potentiometric membrane elec-
equivalence point or when extraneous colored materi- trodes. Anal. Chem. 1982, 54, 1194A.
als are present in the sample. Titrations in solutions Serjeant, E.P. Potentiometry and Potentiometic Titrations; John
Wiley: New York, 1984.
of high or low ionic strength and even in non-aqueous Svehla, G. Automatic Potentiometric Titrators; Pergamon Press:
solution are easily performed. Sensitivity of the New York, 1978.
 Tonicity
Jaymin C. Shah
Pfizer Global Research and Development, Groton, Connecticut, U.S.A.

INTRODUCTION irritation is caused by the non-physiological concen-

tration of dissolved solutes coming in contact with
Parenteral formulations, both large and small volume, sensitive tissues. Tonicity is a formulation property
have been discussed in depth in Volume 11 of the that has a direct influence on the ability of the formu-
Encyclopedia of Pharmaceutical Technology (V 11, lation to result in tissue irritation, as described by the
pp. 201–217, 217–237). However, no discussion of par- following example.
enteral formulations is complete without an adequate If a small quantity of blood defibrinated to prevent
description of tonicity. Tonicity is an important factor clotting is mixed with a solution containing 0.9% w/v
in the formulation of products intended for application of NaCl, the red blood cells remain intact and retain
to sensitive mucous membranes of organs such as eye, their normal size and shape. The NaCl solution is
ear, and nose. In this article, an attempt is made to first considered to be isotonic and has essentially the same
Technology–

introduce tonicity with respect to its physiological salt concentration as does the red blood cell. In con-
Tooling

significance, followed by a discussion of the physico- trast, if the blood is mixed with 1.8% w/v NaCl
chemical basis for tonicity and colligative properties. solution, erythrocytes shrink and become wrinkled or
Then, a brief review of methods of measuring and/or crenated as if the cell content has been sucked out.
calculating tonicity is given, followed by the estab- The salt solution that causes this is considered hyper-
lished methods of adjusting tonicity and the examples tonic with respect to the red blood cell contents. It is
illustrating each of the methods. Tables listing the because the red blood cell contains a lower salt concen-
established values necessary to do the calculations tration than the surrounding 1.8% w/v salt solution
are provided at the end of the article. Excellent com- and as if the water from the erythrocytes passes
prehensive reviews dealing with various aspects of through the cell membrane to dilute the surrounding
tonicity are available in the literature in the form of salt solution to equalize the two salt concentrations
chapters in various textbooks for pharmacy students across the membrane. The opposite phenomenon
(Remington’s Pharmaceutical Sciences, Physical Phar- occurs if blood is mixed with 0.45% w/v NaCl solution.
macy by Martin, etc.). A list of such reading material is Water from the surrounding salt solution enters the
provided at the end of the article in the Bibliography. erythrocytes, causing them to swell and finally burst,
Dosage forms are drug-delivery systems designed to with the liberation of hemoglobin. The 0.45% w/v salt
deliver drug to the systemic circulation or to a localized solution is considered hypotonic, and the phenomenon
region of the human body. These dosage forms should is known as hemolysis. The physiological significance
ideally be free of any undesired adverse effects from of hemolysis was reconfirmed recently by the report
the drug and from the formulation components. of 10 episodes of hemolysis among patients who
Reasonable risks associated with the drug substance received hypotonic 25% human albumin because of
are sometimes tolerated with an objective of realizing dilution with sterile water instead of isotonic sodium
significant therapeutic advantages, as in the case chloride.[1] Two of these 10 recipients exhibited signifi-
of cancer chemotherapeutic agents. However, any cant hemolysis and adverse pathological conditions,
untoward side effect, even as minor as irritation, result- with one resulting in death.[1] Also, it has been
ing from an excipient or the finished dosage form observed that hypertonic and hypotonic salt solutions
cannot be accepted and should not be tolerated. This tend to irritate sensitive mucous membranes of the
concern is particularly important to parenteral formu- eye, the nose, and the muscle when applied. However,
lations that breach the normal defensive barriers of the an isotonic solution causes no tissue irritation when
human body to deliver the drug. Therefore, any formu- it comes in contact with the tissue. The crenation and
lation that comes in contact with sensitive mucous the hemolysis of red blood cells in hypertonic and
membranes of organs such as the eye should not result hypotonic salt solution, respectively, can be explained
in tissue irritation and pain attributable to the formu- by the movement of water across the cell membrane.
lation itself. One of the physicochemical means by A membrane is defined as semipermeable if it allows
which a formulation may result in pain and tissue only the movement of solvent molecules across it.
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-100000443
3768 Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved.
Tonicity 3769

The process of diffusion of a solvent through a semi- because they are all interrelated. The theory of the
permeable membrane from a less-concentrated solu- colligative properties has been well-established and
tion to a more-concentrated solution is known as reviewed in depth in the various textbooks and in the
osmosis. The pressure that must be applied to the con- pharmaceutical literature (see the Bibliography). And
centrated solution side of the membrane to prevent because the focus of this Encyclopedia is on pharma-
the flow of pure solvent across the membrane from the ceutical technology rather than on the theoretical foun-
diluted solution is known as the osmotic pressure. In dation, we briefly overview the theory of colligative
crenation, water diffuses from the inside of the erythro- properties as needed to understand the concepts
cyte across the membrane into the exterior hypertonic used in methods to adjust tonicity of parenteral and
salt solution. Hemolysis occurs when water diffuses ophthalmic formulations.
from the exterior hypotonic solution into the erythro-
cyte, causing it to swell and burst. An isotonic solution
is an aqueous solution that generates the same ‘‘tone,’’ RAOULT’S AND HENRY’S LAWS AS BASIS
or osmotic pressure, as the body fluids across biologi- FOR COLLIGATIVE PROPERTIES
cal membranes and thus prevents water flow in either
direction and hence is non-irritating when injected, Raoult’s law states that in an ideal solution, the partial
instilled, perfused, or brought into contact with sensi- vapor pressure of each volatile constituent is equal to
tive mucous membranes. When a solution is hypertonic the vapor pressure of the pure component multiplied
or hypotonic, osmotic water flow occurs and tone of by its mole fraction in the solution. Thus, for two
the membrane is affected. Thus, formulators need to constituents A and B in solution:
adjust the tone, or tonicity, of the solution to be iso-

Technology–
tonic with physiological fluids. To be able to adjust PB ¼ PBo XB ð1Þ

Tooling
the tonicity of a formulation to the isotonic state,
one has to understand the principles behind the gener- PA ¼ PAo XA ð2Þ
ation of the osmotic pressure resulting from the
dissolved solutes and how it can be altered to that of where PA and PB are the partial vapor pressures of
the physiological fluids. constituents A and B over their solution when the frac-
Osmotic pressure is a colligative property unlike the tional molar concentrations are XA and XB, and the
additive and constitutive properties of solution. Simply vapor pressure of the pure constituents are PoA and PoB.
stated, the colligative properties of a solution are Therefore, it can be inferred that the vapor pressure
dependent solely on the number of non-solvent (solute) of B above the solution by dilution with A is reduced
particles (molecule/ions) dissolved or in a true solution relative to its vapor pressure in pure state and vice
form in a given solvent and are independent of the spe- versa for A. This diminishes the escaping tendencies
cific physicochemical characteristics of the non-solvent of each component, leading to a reduction in the rate
dissolved substance(s). For example, two non-electrolyte of escape of the molecules of A and B from the surface
solutes A and B when prepared as 0.1 M solutions of the solution. This law is valid only in ideal solutions
will exhibit the same osmotic pressure irrespective of in which there are no intermolecular interactions
the chemical nature of A and B. Experimentally, it between components A and B (adhesive interactions)
has been found over the years that the colligative or in which interactions between the two components
properties are indeed independent of the solute nature A and B are identical to the interactions of the pure
and dependent solely on the number of independent components A and pure B (cohesive interactions
particles in dilute solutions for a wide variety of between A and A and between B and B). In essence,
solutes, provided the number of particles is properly the molecule of each component sees an environment
assessed. An implicit assumption in the statement identical to its molecular environment in the pure state.
above is that the solute is non-volatile relative to This refers to an infinitely dilute solution in which a
the solvent and the reasoning will be clear from the component’s thermodynamic activity is equal to its
discussion that follows. The colligative properties of concentration. However, in the real solutions, the
solution are vapor pressure lowering, boiling-point assumption noted above may not apply, and negative
elevation, freezing-point depression, and osmotic deviation from Raoult’s law may occur when adhesive
pressure. These four properties are effects of solute attractions between A and B are greater than cohesive
on the solvent, in that it reduces the escaping tendency attraction within pure A or pure B molecules; i.e., the
of the solvent, and all of them can be related to vapor vapor pressure of the solution or the partial vapor
pressure lowering of the solution. Osmotic pressure is pressure of each component is lower than that expected
of primary importance from the formulation stand- based on Raoult’s law applied to ideal solution. Simi-
point; however, it is cumbersome to measure, and larly, positive deviations from Raoult’s law can occur
therefore other colligative properties are determined when interactions between A and B are less than the
 3770 Tonicity

cohesive interactions of pure A or pure B, resulting in from Raoult’s and Henry’s laws is that the thermody-
vapor pressures higher than that expected based on namic activity of a solvent as measured by its vapor
Raoult’s law applied to ideal solution. In general, pressure is proportional solely on the mole fractional
Raoult’s law states that when a component A is diluted composition of the solute, irrespective of the physical
with another component B, the partial vapor pressure and chemical nature of the dissolved species. The
of A is reduced; in essence, a dilution effect. vapor pressure of the solvent above the solution thus
Raoult’s law does not apply over the entire concen- depends solely on the number of particles (molecules/
tration range in a non-ideal, real solution. However, ions) of the dissolved solute and not on the weight
when one component is in a large enough excess to be concentration of the solute in solution. Therefore, the
considered a solvent, Raoult’s law may be expressed as: vapor pressure of a solvent above a dilute solution that
obeys Henry’s law is a colligative property of the solu-
o
Psolvent ¼ Psolvent Xsolvent ð3Þ tion. Henry’s law has been found to be applicable to
non-electrolyte-type solute mole fractional concentra-
tions of 0.1; however, the range is much smaller for
in such a dilute solution and is valid only for the electrolyte type solutes owing to the long-range nature
solvent component of a non-ideal solution that is suffi- of interionic interactions as noted above. Its impact is
ciently dilute for the other component, i.e., the solute in evaluated later below.
a dilute non-ideal solution. In such a dilute solution, the
solute molecule is completely surrounded by solvent
molecules such that the solute molecule can interact
COLLIGATIVE PROPERTIES
only with the solvent molecules because there are very
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few solute molecules. Further dilution beyond this

Raoult’s law forms the basis for the colligative proper-
Tooling

point does not alter a solute molecule’s environment

ties, and Henry’s law sets the limits of the applicability
and, even if the solute molecule interacts with the sol-
of Raoult’s law to colligative properties of a solution
vent molecule, the solute’s partial vapor pressure or
as increasing amounts of solute are added to solution.
the thermodynamic activity becomes proportional to
As noted above, colligative properties are a conse-
its fractional molar composition as:
quence of the number of dissolved particles in solution
and are all related to the escaping tendency of the
Psolute ¼ Ksolute Xsolute ð4Þ
solvent from solution.
The four colligative properties that are of impor-
Eq. (4) is known as Henry’s law, and Ksolute is the
tance are: 1) the vapor pressure lowering; 2) the
Henry’s law constant, which is less than Posolute. There-
elevation of boiling point; 3) the freezing-point
fore, Henry’s law applies to the solute in dilute solu-
depression; and 4) the osmotic pressure. An attempt
tions, and Raoult’s law applies to solvent in dilute
is made below to describe qualitatively and quantitat-
non-ideal solutions. Note the similarities between
ively each colligative property of solutions, with an
Eqs. (1) and (2) and between Eqs. (3) and (4) for the
emphasis on their interrelationship and their appli-
non-ideal dilute solution case. When the solution is
cation later in measurement and adjustment of the
ideal, Henry’s law becomes identical to Raoult’s law,
tonicity of solutions, with particular reference to par-
and Ksolute becomes identical to Posolute. When the par-
enteral formulations. Although theoretical derivations
tial pressures of the solute and the solvent are directly
based on thermodynamics can be used to show how
proportional to their molefractions over the entire
each of the colligative properties of solution arises
range, the solution is ideal. In a non-ideal solution,
and relate to each other, textbooks on physical chem-
Raoult’s law will apply to the solvent over the entire
istry for theoretical derivations are recommended.
concentration range, whereas Henry’s law will apply
to the solute in a limited concentration range in which
it is in a sufficiently diluted form. Lowering of Vapor Pressure
When a non-volatile solute is dissolved in a solvent,
the partial vapor pressure of the solvent above the The addition of a non-volatile solute to a solvent leads
solution is equal to the vapor pressure of the solution. to a reduction in the vapor pressure of the solvent
And because the mole fraction of the solvent is because of a reduction in thermodynamic activity of
Xsolvent ¼ 1
Xsolute, Eq. (3) can be rewritten as: the solvent. Also, because the solute is non-volatile,
the vapor pressure of the solvent is the vapor pressure
o
Psolution
Psolvent ¼ Psolvent ð1
Xsolute Þ ð5Þ of the solution, as seen from Eq. (5). Qualitatively, one
can imagine that fewer numbers of solvent molecules
for the dilute solution of a non-volatile solute of mole are escaping per unit surface area of the solution than
fraction Xsolute. Therefore, the important conclusion from the pure solvent because fewer solvent molecules
Tonicity 3771

are present per unit surface area of the solution owing lowering based on the Clausius–Clapeyron equation,
to displacement by solute molecules. However, these resulting in an equation relating it to molality as follows:
solute molecules will not affect the condensation of sol-
vent molecules with insufficient kinetic energy present ðTsolution
Tbo Þ ¼ dTb ¼ DTb ¼ K DP ¼ Kb m ð8Þ
in the vapor phase. The result is a net reduction in
escaping tendency of solvent molecules on the surface, where in Kb is called the molal elevation constant or
causing a lowering of vapor pressure and, conse- the ebullioscopic constant, a characteristic constant
quently, the rate of vaporization. The resulting vapor for each solvent, and is the boiling point elevation of
pressure lowering (Posolvent
Psolution) and the relative an ideal 1-molal solution of a non-volatile solute. Kb
vapor pressure lowering as a function of mole frac- can beobtained by measuring the dTb/m of several
tional concentration of solute can be obtained by molal concentrations of solute in solutions and extra-
rearranging Eq. (5) to Eq. (6): polating the dTb/m versus molality curve to zero
solute concentration. The value of Kb for water is
o
Psolvent
Psolution DP 0.515 kg/mole. By measuring the boiling point eleva-
o ¼ o ¼ Xsolute ð6Þ
Psolvent Psolvent tion of a solvent in a solution and knowing the Kb for
that solvent, one can calculate the molal concentration
The left term is the relative vapor pressure lowering, of a solute in the solution. From Eq. (8), it is evident that
which is solely dependent on the mole fraction concen- the elevation of boiling point is a colligative property like
tration of a single solute or the sum of mole fraction of vapor pressure lowering because it is strictly dependent
each solute dissolved in the solution. Thus, the relative on the molal concentration of the solute: the number
vapor pressure lowering is a direct measure of the total of particles in solute and, thus, independent of the

Technology–
number of dissolved solute particles, irrespective of physicochemical nature of the solute.

Tooling
their physicochemical nature. The mole fractions can
be converted into molality (m moles of solute per
1000 g of solvent) to result in the following equation Freezing-Point Depression
for water as the solvent:
The freezing point of a liquid or the melting point of a
DP m Msolvent solid phase of a pure compound is the temperature at
¼ Xsolute ffi
o
Psolvent 1000 which the solid and liquid phases are in equilibrium
at a pressure of 1 atm. The freezing point of a pure
ffi 0:018 m for Aq: solutions ð7Þ
compound is described by a unique point in the phase
diagram of the compound, and, at that point, the solid
where m is the concentration of solute expressed in
and liquid phases are in equilibrium, and the vapor
molality, and Msolvent is the molecular weight of the
pressure of the liquid phase coincides with the vapor
solvent in grams. For water, M ¼ 18, and because
pressure of the pure solid phase. Because in a solution,
the density of water is close to 1, for dilute aqueous
the vapor pressure of the solvent is lowered relative to
solutions, the molality and molarity (moles/liter) can
the pure solvent, no freezing (or crystallization) takes
be used interchangeably, and Eq. (7) can be used to
place at the equilibrium temperature of the liquid
calculate the relative vapor pressure lowering from
and solid phases of the pure solvent; i.e., the freezing
the molar concentration of the non-electrolyte solute.
point. The phase with the lower vapor pressure is the
more stable phase thermodynamically. Therefore,
cooling of the solution below the freezing point of
Elevation of the Boiling Point the pure solvent results in a greater reduction in vapor
pressure of the pure solid phase than the solution
The boiling point of a liquid is the temperature at phase, and when the vapor pressure of the two phases
which the vapor pressure of the liquid becomes equal eventually coincides, freezing (crystallization) of the
to the external pressure acting on the liquid, which is pure liquid solvent occurs. The dissolved solute reduces
760 mm Hg at one atmospheric pressure. Therefore, the the escaping tendency of the solvent molecules to crys-
boiling point of a solution of non-volatile solute will be tallize, and thus the temperature must be reduced to
higher than that of the pure solvent owing to the solute reestablish equilibrium between the solid and liquid
reducing the vapor pressure of the solvent above the phases and hence the depression of freezing point. It
solution according to Raoult’s law. The solution has must be noted that the dissolved solute only affects
to be heated to a higher temperature to achieve the the freezing of the liquid phase and does not alter the
same vapor pressure to result in boiling of the solvent. tendency of the molecules to leave the solid phase,
The elevation of the boiling point (Tsolution
Tbo) is although both processes are occurring in equilibrium
directly proportional to the relative vapor pressure at the freezing point. Also, the solvent should form a
 3772 Tonicity

pure solid; if the solute cocrystallizes with the solvent, Alternatively, one can see that at the beginning, the
the phenomenon is complex and cannot be described solution and pure solvents have different thermo-
as a colligative property. The more concentrated the dynamic activities for the solvent because they have
solution, the greater the freezing-point depression, different vapor pressures. For the solution and pure
and using thermodynamic principles, Raoult’s law, solvent on two sides of the semipermeable membrane to
and Clausius–Clapeyron equation, the freezing-point be in equilibrium, they should have identical escaping
depression can be related to solute concentration tendency or identical vapor pressure and, thus identical
expressed in molality as follows: thermodynamic activity. The equilibrium is therefore
established by the generation of the osmotic pressure
ðTsolution
Tfo Þ ¼ dTf ¼ DTf ¼ K DP ¼ Kf m that compensates for the difference in solvent concen-
ð9Þ tration on the two sides of the membrane, which is
responsible for the different vapor pressures, escaping
tendencies, and thermodynamic activity. Therefore,
where in Kf is called the molal depression constant or
the cryoscopic constant, a characteristic constant for osmosis is a process to reach equilibrium state whereby
each solvent dependent on the physicochemical nature solvent spontaneously flows from the high-free-energy
(low-vapor-pressure) side of the membrane to the low-
of the solvent, and is the freezing-point depression
free-energy (high-vapor-pressure) side of the mem-
of an ideal 1-molal solution of a non-volatile solute.
brane, until the solvent’s free energies on both sides
Kf can be obtained experimentally by measuring the
of the membrane are equal and identical. Obviously,
dTf/m of several molal concentrations of solute in
the solute will not be able to attain equilibrium because
solutions and extrapolating the dTf/m versus molality
it cannot diffuse through the semipermeable mem-
curve to zero solute concentration. The value of Kf for
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water is 1.86 kg/mole. By measuring the freezing-point brane. Because osmotic pressure is attributable to the
Tooling

depression of a solvent in a solution and knowing the Kf difference in vapor pressure of the solvent above solu-
tion, it is also a colligative property as explained below.
for that solvent, one can calculate the molal con-
Using thermodynamics and considering free energy
centration of a solute in the solution. Freezing-point
of the solvent as a function of vapor pressure, the
depression is a colligative property as seen from Eq. (9),
osmotic pressure (p) that develops when a solution is
because it is proportional to molal concentration of
separated from pure solvent by semipermeable mem-
solute: the number of particles in solution, and not on
brane can be related to vapor pressures as shown below:
the physicochemical characteristics of the solute.
o
RT Psolvent
Osmotic Pressure P ¼ M ln ð10Þ
Vsolvent Psolution

As described in the Introduction, the process of

where in R is the gas constant, T is temperature, VM
solvent
diffusion of a solvent through a semipermeable
is the partial molar volume, (the volume occupied by
membrane from a less-concentrated solution into a
1 mole of solvent), and p is the developed osmotic
more-concentrated solution is osmosis. This results in
pressure when a solution with vapor pressure Psolution
the development of a hydrostatic pressure head on
is separated from a solvent with vapor pressure Posolvent
the more-concentrated solution side of the membrane.
by a semipermeable membrane. Applying Raoult’s law
Alternatively, pressure may be applied to the more-
and substituting mole fractions for the vapor pressures
concentrated solution side of the semipermeable mem-
from Eq. (5) into Eq. (10) results in the following:
brane to prevent the diffusion of solvent. This applied
pressure on the concentrated solution is identical to the
RT RT
hydrostatic pressure head that may develop owing to P ¼ M lnð1
Xsolute Þ ffi M Xsolute
Vsolvent Vsolvent
osmosis. It is known as the osmotic pressure and is
since lnð1
Xsolute Þ ffi
Xsolute ð11Þ
directly proportional to the solute concentration in
an ideal solution. A semipermeable membrane is one
that allows the movement of only solvent molecules, In a dilute solution, Xsolute is approximately equal to
and if the membrane is not semipermeable, osmosis the molar ratio nsolute/nsolvent, and Eq. (11) becomes:
may not be observed because the solute will diffuse nsolute nsolute
quickly through the membrane to equalize the concen- P ¼ M RT ¼ RT ¼ mRT
nsolvent Vsolvent Vsolution
tration on two sides of the membrane.
Osmosis tends to equalize the escaping tendencies of ð12Þ
the solvent on both sides of the semipermeable mem-
brane. Escaping tendency can be measured in terms in which the number of moles of solvent multiplied by
of partial vapor pressure of solvent above the solution. the partial molal volume is equal to the volume of the
Tonicity 3773

solvent in solution. In a dilute solution, the volume of slowly across synthetic and semisynthetic polymeric
solvent can be approximated to the volume of solution, membranes, across which the diffusion is very slow.
which results in the above equation relating osmotic
pressure to molar or molal concentration of a solute
in solution. Eq. (12) is known as Morse’s expression COLLIGATIVE PROPERTIES OF
and demonstrates how osmotic pressure is a colligative ELECTROLYTES AS COMPARED
property directly proportional only on the number of WITH NON-ELECTROLYTE SOLUTES
particles dissolved in the solvent irrespective of the nat-
ure of the solute. van’t Hoff had recognized early on The colligative properties, by definition, should be
that there is a direct proportionality among osmotic independent of the nature of the solute. Therefore,
pressure and concentration of solute and temperature, 0.1-molal solutions of sucrose and NaCl should exhibit
and suggested a relationship that was similar to the similar colligative properties. It was observed by van’t
equation for an ideal gas as follows: Hoff that colligative properties of dilute solutions of
non-electrolytes such as sucrose were expressed satis-
PVsolution ¼ nsolute RT ð13Þ factorily by the equations above. However, solutions
of strong electrolytes such as salts gave osmotic pres-
Eq. (13) is analogous to the ideal gas equation, and sure twice or three times as large as would be expected
van’t Hoff concluded that osmotic pressure of a dilute based on Eq. (14), depending on the electrolyte investi-
solution was a pressure that the solute would exert if it gated. To account for this anomaly, van’t Hoff
behaved like a gas occupying that volume. Eq. (13) can proposed the following modification of Eq. (14) as
also be expressed as: shown below:

Technology–
Tooling
nsolute P ¼ i c RT ð15Þ
P ¼ RT ¼ cRT ð14Þ
Vsolution
in which i can be considered to be a factor to account
which shows that osmotic pressure is directly pro- for the deviation of concentrated solutions of electro-
portional to the concentration of solute expressed in lytes and also non-electrolytes from Raoult’s law as
molarity. This equation is similar to Morse’s applied to ideal solutions. After Arrhennius developed
expression, Eq. (12); however, it has been shown theo- the theory of ionization or dissociation of salts into
retically and experimentally that more accurate results ions, van’t Hoff and others recognized that the value
can be obtained when solute concentration is expressed of i approached or equaled the number of ions into
in molality rather than in molarity. Although the which the electrolyte or the molecule dissociated as
resemblance of Eq. (13) to the ideal gas equation is the solution was made more dilute. For example, a
striking, osmotic pressure is a result of differences in dilute 0.1 M solution of NaCl would be twice as active
the escaping tendencies of the solvent on two sides of osmotically as a 0.1 M solution of sucrose, and i for
the membrane rather than of the behavior of a solute NaCl was two. Similarly, 0.1 M solutions of CaCl2
such as a gas. and MgCl2 would generate three times the osmotic
From Eqs. (12)–(14), one finds that 1-molar solution pressure of 0.1 M sucrose solution, and i for both is
of any solute will generate an incredibly high osmotic equal to three. Therefore, it was realized that i reflected
pressure of approximately 24 atm at room tempera- the number of ions the electrolytes dissociated into
ture, which has been verified experimentally. Although and, thus, electrolytes at equimolar concentrations
this estimate is based on the assumption that the were more effective in generating osmotic pressure
solution is dilute and behaving ideally, at high concen- based on the number of ions they produced on dis-
trations of solute, the theory overestimates the experi- sociation. However, it was also observed that at
mental findings. The discussion above deals primarily moderate concentrations of electrolytes, osmotic pres-
with the thermodynamic basis for the generation of sures were less than that expected based on complete
osmotic pressure; however, it does not address the dissociation. In fact, this led the scientific community
issue of how fast the equilibrium is attained or how fast to suggest partial dissociation for even strong electro-
the osmotic pressure will be generated. The rate of gen- lytes and to use colligative properties as a measure of
eration of osmotic pressure is a kinetic process and degree of dissociation. However, we know now that
depends to a great extent on the characteristics of the strong electrolytes do dissociate completely even in
semipermeable membrane. Red blood cell membrane concentrated solutions from other measurements such
or the mucous membrane in the eye are very thin as conductivity techniques. The lower-than-expected
and moist, and water can diffuse very rapidly through values of osmotic pressure in moderate concentra-
the membrane to generate the enormous osmotic pres- tions of electrolytes is attributable to the influence of
sure. However, the osmotic pressure may develop very long-range ionic interactions that come into play as
 3774 Tonicity

the solution gets increasingly concentrated. The basic than does the surrounding hypertonic 1.8% w/v salt
assumption in Raoult law was that there was no inter- solution, and the water inside the cells diffuses through
action between solute particles and, even if there was the cell membrane to dilute the surrounding salt
any, it should equal that between the solute and the solution to equalize the osmotic pressure across the
solvent. However, the strong attractive forces between membrane. The exact opposite phenomenon occurs if
ions of opposite charges do predominate in increas- blood is mixed with hypotonic 0.45% w/v NaCl sol-
ingly concentrated solutions of electrolytes, and their ution. Water from the surrounding salt solution enters
thermodynamic activity is reduced relative to that in the cells, causing them to swell and finally burst with
an infinitely dilute solution. Also, the effect of ionic the liberation of hemoglobin, and the phenomenon is
strength of a solution has been shown to influence known as hemolysis. The crenation and hemolysis of
the activity of electrolytes, and thus, it is the interionic red blood cells in hypertonic and hypotonic salt solu-
forces rather than the partial dissociation that seems to tion, respectively, are explained by the movement of
influence the colligative properties of electrolytes being water across the cell membrane owing to the osmotic
lower than expected based on complete dissociation. pressure differential. However, it is well-known that
All the colligative properties of all solutes with the different physiological membranes are different
the modification by the van’t Hoff factor i can be with respect to their permeability characteristics. The
expressed as: red blood cell membrane has been found to be per-
meable to small polar and semipolar solutes such as
o
DP ¼ i Psolvent m; P ¼ i RT m; alcohol, boric acid, and urea, etc. Thus, the erythrocyte
DT f ¼ i Kf m; DTb ¼ i Kb m ð16Þ membrane is not truly semipermeable, and although
2% boric acid solution is iso-osmotic with erythrocyte
Technology–

cell contents, it causes hemolysis because boric acid

Tooling

where in i for non-electrolyte should be 1 and for

strong electrolytes equal the number of ions formed moves freely across the membrane and its solution
being hypotonic acts like pure water in its effect on
on dissociation. For example, i should be 2 for NaCl,
erythrocytes. However, the same 2% boric acid solu-
3 for CaCl2, and 4 for FeCl3. However, in reality, i is
tion is both iso-osmotic and isotonic with eye secre-
less than that calculated, based on the number of ions
tions and causes no irritation when instilled in the
produced in concentrated solutions, but will approach
eye because the mucous membrane of the eye is a true
the theoretical number in infinitely dilute ideal solu-
semipermeable membrane. To resolve the confusion
tions. When the i value is calculated, for a number of
created by different permeability characteristics of bio-
solutions with increasing concentration of the solute
and then extrapolated to zero concentration of the logical membranes, the word isotonicity was created.
The word isotonic refers to solutions that are iso-
solute, one can obtain the theoretical i value. The van’t
osmotic with the cell contents, across a specific
Hoff factor has also been considered the ratio of any
membrane, and in addition, maintains the tone of the
colligative property of a real solution to that of an
membrane, i.e., no solvent movement across the mem-
ideal solution of a non-electrolyte.
brane. Thus, 2% boric acid solution is iso-osmotic with
blood but it behaves like a hypotonic solution with
erythrocytes while it is both iso-osmotic and isotonic
PHYSIOLOGICAL AND CLINICAL with respect to the mucous membrane of the eye.
SIGNIFICANCE OF TONICITY It has been also observed that hypertonic and hypo-
tonic salt solutions tend to irritate sensitive tissue and
Osmotic pressure becomes important from a physio- cause pain when applied to mucous membranes of the
logical standpoint because a majority of biological eye, ear, and nose, etc., whereas isotonic solution
membranes are semipermeable, and body fluids such causes no tissue irritation when it comes in contact
as blood and tears exhibit significant osmotic pressure with the tissue. Obviously, the tonicity of formulations
owing to a number of solutes dissolved in them. As that come in to direct contact with blood, muscle, eye,
noted above in the Introduction, if a small quantity of nose, and delicate tissues is critical. Therefore, the issue
blood is mixed with a solution containing 0.9% w/v of tonicity is important in small- and large-volume
NaCl, the red blood cells remain intact and retain their injectables, ophthalmic products, and products
normal size and shape. The NaCl solution is con- intended for tissue irrigation. The degree of tissue irri-
sidered to be isotonic because it maintained the tone tation or hemolysis or crenation observed depends on
of the membrane of the red blood cell. In contrast, if the degree of deviation from isotonicity, the volume
the blood is mixed with the hypertonic 1.8% w/v NaCl injected, the speed of injection, the concentration of
solution, cells shrink and become wrinkled or crenated the solutes in the injection, and the nature of the
owing to its content being sucked out. It is because the membrane. The parenteral and ophthalmic formula-
red blood cell content exerts a lower osmotic pressure tions are therefore adjusted to isotonicity if possible.
Tonicity 3775

Hypotonic solutions can be easily adjusted to isoton- however, one has to be mindful of the fact that solu-
icity by adding solutes such as dextrose or sodium tions may be iso-osmotic with erythrocyte contents,
chloride, commonly used for this purpose. However, yet may cause hemolysis because solutes such as boric
at times, the formulation may be hypertonic and may acid are permeable through erythrocyte membrane
have to be diluted with water to maintain isotonicity. and, thus, solution is not isotonic.[2] Therefore,
This dilution of the hypertonic solution may be pre- Grosicki and Husa recommended that the word iso-
cluded owing to other limitations such as poor aqueous tonic should be used with reference to solutions having
solubility of the drug. In such a case, the hypertonic equal osmotic pressures with respect to a particular
solution can be administered slowly in small volumes membrane.[2] Because hemolysis due to hypotonic
into a large vein such as the subclavian vein in which solution results in release of oxyhemoglobin directly
the formulation will be diluted and distributed rapidly, proportional to the number of cells hemolyzed, a
minimizing chances of crenation of erythrocytes, pain, quantitative method has been developed to calculate
and tissue irritation on injection. It has also been osmotic pressure and the van’t Hoff i factor noted
observed that minor deviations such as 10% from above. A limitation of observing changes in erythro-
isotonicity may result in no effect or only temporary cytes as a measure of tonicity is the fact that the
effects at the site of injection. However, the effects of specific chemical interaction of the solute with the cell,
deviation from isotonicity of large-volume parenterals pH of the solution, presence of solvents, lipid solubility
can be fairly severe and, thus, parenteral nutrient solu- of the solute, and denaturant activity of solute may
tions and infusions of large volume need to be adjusted have influences on the cell membrane and, thus,
to isotonicity. Large-volume infusion of hypotonic osmotic pressure differences alone are not responsible
solutions has been observed to cause effects ranging for hemolysis. Furthermore, it was shown recently that

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from hemolysis to water-retention problems such as hemolysis is related to the contact time in addition to

Tooling
convulsions and pulmonary edema. This was exempli- hypotonicity of the formulation.[3] To overcome this
fied by the recent report of 10 episodes of hemolysis, limitation, some investigators have used measurements
with two patients exhibiting significant hemolysis and of erythrocyte cell volumes as a function of tonicity of
renal insufficiency resulting in one death. These severe solution, which influence solvent (water) uptake or loss
episodes of hemolysis occurred because of the large from erythrocytes. This method is more sensitive,
volume infusion of 25% human albumin diluted with objective, and reliable than observation of hemolysis.
sterile water instead of with isotonic sodium chloride Recently, a method using fluorescence anisotropy for
for therapeutic plasma exchange.[1] In contrast, large- fluidity of erythrocyte membranes demonstrated differ-
volume infusion of hypertonic solutions can result in ences between hypotonic and isotonic solutions.[4]
severe conditions such as intracellular dehydration, However, the method is involving, and more data need
osmotic diuresis, hyperglycemia, glycosuria, dehy- to be obtained to correlate tonicity with fluidity of the
dration from loss of water, and coma. Also, hypertonic membrane to be reliable.
solution infusion should be terminated gradually to An alternative approach is based on the theoretical
avoid sudden changes in osmotic pressure. In sum- foundation described earlier for the colligative proper-
mary, any formulation that comes in to contact with ties. If the solution is isotonic with blood, its osmotic
sensitive tissues of the human body needs to be pressure, vapor pressure, boiling-point elevation, and
adjusted to isotonicity to minimize any adverse effects. freezing-point depression should also be identical to
To be able to adjust the formulation to isotonicity, those of blood. Thus, to measure isotonicity, one has
a method to measure the tonicity and/or the osmotic to measure the osmotic pressure of the solution and
pressure of the formulations has to be used. The compare it with the known value for blood. However,
methods to measure tonicity or osmotic pressure of the accurate measurement of osmotic pressure is diffi-
solutions are reviewed briefly as below. cult and cumbersome. If a solution is separated from
blood by a true semipermeable membrane, the result-
ing pressure due to solvent flow (the head) is accurately
MEASUREMENT OF TONICITY measurable, but the solvent flow dilutes the solution,
OF SOLUTIONS thus not allowing one to know the concentration of
the dissolved solute. An alternative is to apply pressure
The most direct method for measurement of tonicity to the solution side of the membrane to prevent
obviously would be to observe changes in erythrocytes osmotic solvent flow. In 1877, Pfeffer used this method
on mixing solution with blood. If hemolysis or crena- to measure osmotic pressure of sugar solutions. With
tion or a marked change in the appearance of erythro- the advances in the technology, sensitive pressure
cytes occurs, the solution is not isotonic. If the cells transducers, and synthetic polymer membranes, this
retain their normal size and shape, the solution is iso- method can be improved. However, results of the
tonic. Grosicki and Husa used this method early on; search for a true semipermeable membrane are still
 3776 Tonicity

elusive, and this method is still cumbersome and incon- in contact with solution (aq.) and the freezing point
venient. The measurement of osmotic pressure using measured and compared with that of the pure solvent
this method has been applied successfully to colloidal (water) in contact with the solid solvent (ice). Also,
solution of proteins to measure their molecular weight one has to consider the presence of other solvents in
because they are of relatively large molecular weight influencing the freezing-point depression. The freezing-
and are impermeable across number of membranes. point depression method is precise to the extent that
Numerous instruments, known as osmometers, are the differences in freezing points of two systems
commercially available to measure osmotic pres- within 0.0002 C can be measured. Freezing-point
sure.[5,6] Only the Knauer membrane osmometer and osmometry can be used for all samples with osmolal-
colloid osmometer are true osmometers using a semi- ities less than 550 mmol/kg, including those that con-
permeable membrane.[5] Vapor pressure osmometers tain volatile solutes.[6] Once the freezing point of the
such as the Wescor osmometer, using the principle of solution is known, inert solute such as sodium chloride
vapor pressure lowering should not be called osmo- is added to match the freezing point of solution with
meters. These types of instruments measure the vapor that of blood and lacrimal fluids. After considerable
pressure using the isopiestic method, or the thermo- debate and experimentation, it is now well-established
electric method, or the measurement of the dew point that
0.52 C is the freezing point of blood and lacri-
of unknown solution in comparison with a standard, mal fluids, following the work of Lund, Nielsen, and
and then calculate osmotic pressure and osmolality Pedersen-Bjergaard.[7] This is also the freezing point
using the theory of colligative properties. These instru- of 0.9% sodium chloride solution, which is therefore
ments require a few microliters, and the method is considered to be isotonic with both blood and lacrimal
fairly precise, simple, and totally automated. The pres- secretions. Therefore, to determine the tonicity of a
Technology–

ence of a volatile solvent such as ethanol will create solution, one has to measure its freezing-point depres-
Tooling

problem with this method because the inherent sion and compare it with that of blood (
0.52 C).
assumption is that only water is present in the vapor The freezing-point depressions of a number of drugs
phase. This can be a serious limitation because many are listed in Table 1. A more comprehensive list of the
parenteral and, ophthalmic formulations contain freezing-point depressions of various concentrations of
organic solvents for the purposes of drug solubility drugs can be found in The Merck Index, Remington’s
and, sometimes of stability. Commercial osmometers Pharmaceutical Sciences, and other literature sources
of this type have been found to measure osmolalities listed in the Bibliography. An important consideration
in the range of 100–3000 mmol/kg reliably. when using this method is that although all the solutes
Boiling-point elevation can also be used to measure present in solution contribute to its freezing-point
osmotic pressure and tonicity of a solution using just a depression, those that permeate the biological mem-
reflux condenser and a thermometer. The commercially brane will not maintain the tone, for example, boric
available instrument is the Cottrell boiling-point appa- acid. In addition, association of solute molecules by
ratus. However, this method is affected by the ambient processes such as complexation and micellar associ-
barometric pressure and the presence of volatile ation, which are temperature-dependent, may have to
solvents in the solution. be considered. The viscosity and presence of suspended
Osmometers based on the freezing-point depression particles can also affect the freezing point by altering
are the most commonly and widely used instruments the crystallization of the solvent. Nevertheless, freez-
for measurements of tonicity because of the simplicity, ing-point depression has become popular because of
reliability, and ease of use. Freezing-point depression its simplicity, reliability, and availability of commercial
of solutions of a number of drugs at various concen- instruments. The methods of osmometry, the techno-
trations has already been determined, and thus, an logy, and the limitations inherent in each method have
extensive database is available for adjustment of been reviewed recently[5,6] and should be consulted for
tonicity of solutions of these drugs, as addressed below. more details.
The freezing-point depression of a solution can be Based on the theory of colligative properties and
simply measured using a salt–ice bath, Dewars flask, the principles of osmometry, it is understood that
and Beckmann’s thermometer. Numerous commercial osmometer will read osmolalities and not osmolarities
instruments requiring small quantities of solution such because colligative properties are directly proportional
as Osmette from Precision Systems that use the prin- to the total solute concentration expressed in molality
ciple of Beckmann’s freezing-point method are now [see Eqs. (1)–(16)]. The relationship between osmolality
available. One of the problems with this method is and osmolarity and its significance can be found in
the disengagement of ice and the need for determi- the Remington’s Pharmaceutical Sciences and in a
nation of the actual equilibrium freezing point. The review article by Deardorff.[10] However, it is more
latter limitation can be overcome by use of the equi- convenient to use osmolarity because it is based on
librium method, in which solid solvent (ice) is placed weight/volume rather than on weight/weight as in
Tonicity 3777

Table 1 Freezing point depressions (Tf1%), Liso values and sodium chloride equivalents (E) for drugs and excipients
for adjusting their solutions to isotonicitya
Substance MWb T1%
f
c
Lisod Ee Vf
Alcohol, dehydrated 46.07 0.41 1.9 0.70 23.3
Aminophylline 456.46 0.10 4.6 0.17 5.7
Amphetamine sulfate 368.49 0.13 4.8 0.22 7.3
Antipyrine 188.22 0.10 1.9 0.17 5.7
Antazoline (Antistine) hydrochloride 301.81 0.11 3.2 0.18 6.0
Apomorphine hydrochloride 312.79 0.08 2.6 0.14 4.7
Ascorbic acid 176.12 0.11 1.9 0.18 6.0
Atropine sulfate 694.82 0.07 5.3 0.13 4.3
Aureomycin hydrochloride 544 0.06 3.5 0.11 3.7
Barbital sodium 206.18 0.29 3.5 0.29 10.0
Benadryl hydrochloride 291.81 0.34 3.4 0.20 6.6
(diphenhydramine hydrochloride)
Boric Acid 61.84 0.29 1.8 0.50 16.7
Butacaine sulfate 710.95 0.12 8.4 0.20 6.7
Caffeine 194.19 0.05 0.9 0.08 2.7
Calcium gluconate 448.39 0.09 4.2 0.16 5.3

Technology–
Calcium lactate 308.30 0.14 4.2 0.23 7.7

Tooling
Camphor 152.23 0.12 1.8 0.20 6.7
Chloramphenicol (chloromycetin) 323.14 0.06 1.9 0.10 3.3
Chlorobutanol (chloretone) 177.47 0.14 2.5 0.24 8.0
Cocaine hydrochloride 339.81 0.09 3.2 0.16 5.3
Dextrose–H2O 198.17 0.09 1.9 0.16 5.3
Dibucaine hydrochloride 379.92 0.08 2.9 0.13 4.3
Ephedrine hydrochloride 201.69 0.18 3.6 0.30 10.0
Ephedrine sulfate 428.54 0.14 5.8 0.23 7.7
Epinephrine bitartrate 333.29 0.11 3.5 0.18 6.0
Epinephrine hydrochloride 219.66 0.17 3.7 0.29 9.7
Fluorescein sodium 376 0.18 6.9 0.31 10.3
Glycerin 92.09 0.20 1.8 0.34 11.3
Homatropine hydrobromide 356.26 0.10 3.6 0.17 5.7
Lactose 360.31 0.04 1.7 0.07 2.3
Magnesium sulfate  7H2O 246.50 0.10 2.5 0.17 5.7
Menthol 156.26 0.12 1.8 0.20 6.7
Meperidine hydrochloride 283.79 0.12 3.7 0.22 7.3
Methamphetamine hydrochloride 185.69 0.22 4.0 0.37 12.3
Morphine hydrochloride 375.84 0.09 3.3 0.15 5.0
Morphine sulfate 758.82 0.08 6.2 0.14 4.8
Naphazoline hydrochloride 246.73 0.16 3.3 0.27 7.7
Neomycin sulfate — 0.06 — 0.11 3.7
Neostigmine bromide 303.20 0.11 3.2 0.22 6.0
Nicotinamide 122.13 0.15 1.9 0.26 8.7
Penicillin G potassium 372.47 0.11 3.9 0.18 6.0
Penicillin G Procaine 588.71 0.06 3.5 0.10 3.3
Penicillin G sodium 356.38 0.11 3.8 0.18 6.0
Phenacaine hydrochloride 352.85 0.11 3.3 0.20 5.3
Phenobarbital sodium 254.22 0.14 3.6 0.24 8.0
(Continued)
 3778 Tonicity

Table 1 Freezing point depressions (Tf1%), Liso values and sodium chloride equivalents (E) for drugs and excipients
for adjusting their solutions to isotonicitya (Continued)
Substance MWb T1%
f
c
Lisod Ee Vf
Phenol 94.11 0.20 1.9 0.35 11.7
Phenylephrine hydrochloride 203.67 0.18 3.5 0.32 9.7
Physostigmine salicylate 413.46 0.09 3.9 0.16 5.3
Physostigmine sulfate 648.45 0.08 5.0 0.13 4.3
Pilocarpine nitrate 271.27 0.14 3.7 0.23 7.7
Potassium acid phosphate (KH2PO4) 136.13 0.25 3.4 0.43 14.2
Potassium chloride 74.55 0.45 3.3 0.76 25.3
Potassium iodide 166.02 0.20 3.3 0.34 11.3
Procaine hydrochloride 272.77 0.12 3.4 0.21 7.0
Quinine hydrochloride 396.91 0.08 3.3 0.14 4.7
Scopolamine hydrobromide 438.32 0.07 3.1 0.12 4.0
Silver nitrate 169.89 0.19 3.3 0.33 11.0
Sodium acid phosphate (NaH2PO4  H2O) 138.00 0.24 3.2 0.40 13.3
Sodium benzoate 144.11 0.24 3.4 0.40 13.3
Sodium bicarbonate 84.00 0.38 3.2 0.65 21.7
Sodium bisulfite 104.07 0.36 3.7 0.61 20.3
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Tooling

Sodium borate  10H20 381.43 0.25 9.4 0.42 14.0

Sodium chloride 58.45 0.58 3.4 1.00 33.3
Sodium iodide 149.92 0.23 3.4 0.39 13.0
Sodium nitrate 85.01 0.39 3.4 0.68 22.7
Sodium phosphate, anhydrous 141.98 0.31 4.4 0.53 17.7
Sodium phosphate  2H20 178.05 0.25 4.4 0.42 14.0
Sodium phosphate  7H2O 268.08 0.17 4.6 0.29 9.7
Sodium phosphate  12H2O 358.21 0.13 4.6 0.22 7.3
Sodium propionate 96.07 0.36 3.4 0.61 20.3
Sodium sulfite, exsiccated 126.06 0.38 4.8 0.65 21.7
Streptomycin sulfate 1457.44 0.04 6.0 0.07 2.3
Strong silver protein — 0.0 — 0.08 2.7
Sucrose 342.30 0.05 1.6 0.08 2.7
Sulfacetamide sodium 254.25 0.14 3.4 0.23 7.7
Sulfadiazine sodium 272.27 0.14 3.8 0.24 8.0
Sulfamerazine sodium 286.29 0.14 3.9 0.23 7.7
Sulfanilamide 172.21 0.13 2.2 0.22 7.3
Sulfathiazole sodium 304.33 0.13 3.9 0.22 7.3
Tetracaine hydrochloride 300.82 0.11 3.2 0.18 6.0
Tetracycline hydrochloride 480.92 0.08 4.0 0.14 4.7
Tripelennamine hydrochloride 291.83 0.17 3.8 0.30 7.3
Urea 60.06 0.35 2.1 0.59 19.7
Zinc chloride 139.29 0.37 5.1 0.62 20.3
Zinc Sulfate  7H2O 287.56 0.09 2.5 0.15 5.0
a
Values vary somewhat with concentration, and those in the table are for 1 to 3% solutions of the drugs in most instances. A more
comprehensive table of E and Tf, values is found in Ref.[8].
b
MW is the molecular weight of the drug.
c 1%
Tf is the freezing point depression of a 1% solution of the drug.
d
Liso is the molar freezing point depression of the drug at a concentration approximately isotonic with blood and lacrimal fluid.
e
E is the sodium chloride equivalent of the drug.
f
V is the volume in milliliters of isotonic solution that can be prepared by adding water to 0.3 g of the drug (the weight of drug in one
fluid ounce of a 1% solution).
(Adapted with modifications from Ref.[9].)
Tonicity 3779

osmolality. The U.S. Pharmacopeia also recommends two types. In class I methods, some inert substance
that the labeling of parenteral and ophthalmic formu- such as sodium chloride or dextrose is added to the solu-
lation should list osmolarity while the experimen- tion to lower its freezing point to match that of blood
tally determined quantity is osmolality. Methods to (
0.52 C) i.e., made isotonic by the addition of inert
convert osmolality to osmolarity using determinations excipient. In class II methods, a calculated quantity
of solution density and solute content[11,12] or using of water is added to the total solute content (drug) of
partial molal volume of solute and solvent have been the prescription to make it isotonic, which is then
described.[13,14] diluted with sufficient isotonic diluting solution to
bring it to the final volume. These methods are
explained below, followed by a simple example illus-
THEORETICAL METHOD TO CALCULATE trating the method. However, the assumptions inherent
TONICITY USING Liso VALUE in all the methods need to be considered carefully. The
first assumption is that colligative properties are addi-
In the discussion above of colligative properties of elec- tive for mixture of solutes and that they are related
trolytes, the equations were modified by introduction linearly to their concentration expressed in molarity,
of the van’t Hoff i factor as shown in Eq. (16). Because molality, or in percentages. This assumption is true
the freezing-point depressions of strong and weak elec- in dilute solutions of non-electrolytes and electrolytes.
trolytes are always greater than those calculated from However, when dealing with concentrated solutions,
Eq. (9), because of different degrees of ionization and this assumption may not be valid. In cases of chemical
interionic interaction, a new factor, L ¼ iKf, is intro- interaction, association, complexation, or micellar
duced. The L value obtained from freezing-point interaction among solutes in solution, the colligative

Technology–
depression of a solution of a particular type of electro- properties of solutes may not be additive. The second

Tooling
lyte at a molar concentration (c) that is isotonic with assumption is that they consider all solutes present in
blood is defined as Liso (Liso ¼ 0.52 /c). For example, solution to be contributing to its tonicity. However,
the Liso value for an isotonic sodium chloride solution it is known from the discussion above that all biologi-
(0.9% w/v) is 3.4, and its freezing-point depression is cal membranes are not truly semipermeable, and thus,
0.52 C. Because the colligative properties are inde- some solutes will not contribute to the tonicity of the
pendent of the chemical nature of the electrolyte solution across that membrane, for example, boric acid
and the interionic interactions in dilute solutions are across erythrocytes membrane. Nevertheless, the errors
similar, all electrolytes of the same type will have introduced are small, and slight deviations from iso-
identical Liso values. Therefore, all non-electrolytes tonicity on either side do not result in significant
have Liso ¼ 1.9, whereas uni-univalent electrolyte’s adverse effects. In the literature, one may also find
Liso ¼ 3.4 and triunivalent electrolyte’s Liso ¼ 6.0. methods known as the L value method or the Liso
Thus, if the ionic nature of the solute and its molecular method, which are identical to the theoretical method
weight are known, using the appropriate Liso value, described above for solutes, for which one can calcu-
freezing-point depression can be calculated for a late the freezing-point depression based on molecular
solution of a given concentration. The average Liso weight and ionic nature. By knowing the freezing-point
values for all types of solutes are available in the litera- depression, any of the class I or class II methods can
ture.[9] This method is simple and does not require then be used to adjust their solution to isotonicity.
experimentation, but it is only approximate, with
potential for some error. Also, one has to know the
ionic nature of the solute, which can be difficult to Class I Methods
determine for a new drug compound with a complex
structure. Freezing-point depression method
(cryoscopic method)

METHODS OF ADJUSTING TONICITY The freezing-point depression of a number of drugs

and excipients, either experimentally determined by
From the theoretical background presented above, the method described above or calculated theoretically
one can easily devise his or her own methods to adjust using the Liso method, is available in the litera-
tonicity of solutions using the principles of colligative ture.[8,9,15] Table 1 lists the freezing point depression
properties. However, in the practice of pharmacy, a of 1% solution of various drugs. Basically, from the
number of simple methods to adjust tonicity of formu- percentage of drug present in solution, the freezing
lation in a prescription order on an extemporaneous point of the solution is calculated. This number is then
basis were developed to help the pharmacist. The subtracted from the freezing point of blood (
0.52 C)
methods of adjusting tonicity could be classified into to obtain the freezing-point depression to be achieved
 3780 Tonicity

by the addition of sodium chloride. Knowing that 0.9% Physostigmine salicylate (2 g/100 ml) is equivalent
sodium chloride is isotonic and freezes at
0.52 C, the to 2  0.16 (E) ¼ 0.32 g/100 ml of sodium chloride.
amount of sodium chloride to be added is calculated Therefore, 0.58 g (0.9–0.32 g) of sodium chloride has to
as shown in the example below. be added to 100 ml of this solution to make it isotonic.
Example 1: Calculate the amount of sodium chlor- Note that the answers given by the two methods are
ide needed to prepare 100 ml of 2% isotonic physostig- not identical, but very close.
mine salicylate solution.
Freezing-point depression of 2% physostigmine sal-
icylate ¼ 2  0.09 C ¼ 0.18 C (Table 1). Therefore, Class II Methods
the freezing-point depression to be achieved by adding
sodium chloride ¼ 0.52 C
0.18 C ¼ 0.34 C. The class II methods involve the calculation of a quan-
Sodium chloride (0.9%) produces a freezing-point tity of water needed to make an isotonic solution for
depression of
0.52 C; therefore, the percentage of a given amount of drug, followed by dilution with
sodium chloride needed ¼ (0.34 C/0.52 C)  0.9% ¼ an isotonic solution to make up the volume. These
0.59% ¼ 0.59 g/100 ml. methods were developed to enable pharmacists to
prepare parenteral and ophthalmic formulations with
simplicity and ease.
Sodium chloride equivalent (E) method

The sodium chloride equivalent (E) is the amount of The White-Vincent method
sodium chloride equivalent to 1 g of the drug in exert-
ing the same osmotic effect. The E value for a new In this method, the weight of the drug (w) is first mul-
Technology–

drug can be calculated from its Liso value or from the tiplied by its sodium chloride (E) to obtain the quantity
Tooling

freezing-point depression as shown below. of sodium chloride osmotically equivalent to weight

The freezing-point depression of a 1 g/L-solution of per gram of drug.[17] Because 0.9 g of sodium chloride
a new drug can be expressed as: dissolved in 100 ml results in an isotonic solution, the
volume of isotonic solution that can be prepared from
1g weight per gram of drug is given by the following
DTf ¼ Liso ð17Þ equation:
MW

By definition, E gram of sodium chloride 100

V ¼ wE ¼ 111:1 w E ð20Þ
(MW ¼ 58.45 and Liso ¼ 3.4) in 1 L will have similar 0:9
freezing-point depression as shown below:
Thus, dissolving weight per gram of drug in V ml of
Eg water will result in an isotonic solution that can be
DTf ¼ 3:4 ð18Þ
58:45 further diluted with isotonic solutions such as 0.9%
sodium chloride or isotonic dextrose solution to make
Therefore, equating Eqs. (17) and (18), results in the up the volume. The method can be illustrated by the
following equation for E: following example.
Example 3: Prepare 100 ml of 2% physostigmine
Liso
E ¼ 17 ð19Þ salicylate solution isotonic with blood.
MW
Using Eq. (20) and E of physostigmine salicylate ¼
Wells developed a nomogram based on the above 0.16 from Table 1, the volume of water needed to
equation to readily calculate E values from the MW prepare isotonic solution, V ¼ 2 g  0.16 
and Liso value of the drug.[16] Thus, the E value for 111.1 ml/g ¼ 35.55 ml. This solution can be diluted
physostigmine salicylate (MW ¼ 413.46) calculated with 64.45 ml of any isotonic diluting solution to
using Liso ¼ 3.4 for a uni-univalent electrolyte is equal obtain 100 ml of 2% isotonic physostigmine salicylate
to 0.14, which is close to 0.16 (E value) in Table 1. This solution. To verify the results, if we assume that we
small deviation is attributable to the difference dilute the above solution with 64.45 ml of isotonic
between the experimentally determined Liso (3.9) of sodium chloride solution, the equivalent amount of
physostigmine salicylate and the theoretical value of sodium chloride added is 0.58 g, which matches with
3.4 for a uni-univalent electrolyte. By knowing the E results obtained using the class I methods.
value, the solution can be adjusted to isotonicity as
shown below. The Sprowls method
Example 2: Calculate the amount of sodium chlor-
ide needed to prepare 100 ml of 2% isotonic physo- In the early days of pharmacy practice, many prescrip-
stigmine salicylate solution. tions were written to prepare one fluid ounce of a 1%
Tonicity 3781

drug solution, thus, the amount of drug (w ¼ 0.3 g) of these systems need to be investigated too. There is
and the final volume were fixed (one fluid ounce or an increasing concern regarding tissue irritation and
30 ml). Sprowls, recognizing this fact, suggested a muscle injury at the site of injection resulting from
modification of the White–Vincent method to further formulations. With the advent of biotechnology, more
simplify the calculations for the practicing pharma- peptide and protein drugs are in clinical trials than
cist.[18] In this method, the amount of drug is fixed at before, and, also, gene therapy is being considered
0.3 g (30 ml of 1% solution), and the volume of water for few diseases. The parenteral formulations of these
required to prepare the isotonic solution is calculated newer drugs are more complex to maintain the integ-
using Eq. (20) and listed in a table such as column 4 rity of their higher-order structure. Therefore, the issue
in Table 1 for all drugs that are commonly used in par- of tonicity needs to be revisited with a newer approach
enteral and ophthalmic formulations and for which and from a different perspective.
sodium chloride equivalents are known. The pharma-
cist then makes up the volume of the preparation to
30 ml with an isotonic diluting solution to fill the
prescription. For example, if one fluid ounce of 1% REFERENCES
physostigmine salicylate solution is to be prepared,
from Table 1, column 4, we recognize that 5.3 ml of 1. Morbidity & Mortality Weekly Report 1999, 48 (8),
157–159.
water is required for 0.3 g of physostigmine salicylate 2. Grosicki, T.S.; Husa, W.J. J. Am. Pharm. Assoc., Sci. Ed.
to prepare an isotonic solution. After the preparation 1954, 43, 632–636.
of this 5.3 ml solution, it can be diluted with any 3. Krzyzaniak, J.E.; Raymond, D.M.; Yalkowsky, S.H. PDA
J. Pharm. Sci. Tech. 1996, 50 (4), 223–226.
isotonic diluting solution to make up the volume to 4. Au, K.S. Biochem. Mol. Bio. Int. 1994, 32, 49–53.

Technology–
one fluid ounce. If one needed to prepare 100 ml of 5. Bevan, D.R. Anesthesia 1978, 33, 794–800.

Tooling
a 1% solution, the volume of water (V) should be 6. Sweeney, T.E.; Beuchat, C.A. Am. J. Physio. 1993, 264,
R469–480.
multiplied by 3.33 to obtain the amount of water 7. Lund, C.G.; Nielsen, P.; Pedersen-Bjergaard, K. The Prep-
necessary to make it isotonic. aration of Solutions Iso-Osmotic with Blood, Tears, and
Tissue; Danish Pharmacopeia Commission: Einar
Munksgaard, Copenhagen, 1947; 2.
8. Budavari, S., O’Neil, M.J., Smith, A., Heckelman, P.E.,
CONCLUSIONS Kinneary, J.F., Eds.; The Merck Index: An Encyclopedia
of Chemicals, Drugs and Biologicals, 12th Ed.; Merck &
Co. Inc.: Whitehouse Station, NJ, 1996; 47–57, MISC.
The theory of colligative properties is well-understood 9. Martin, A.; Bustamante, P.; Chun, A.H.C. Physical phar-
and successfully applied to parenteral formulations for macy: Physical chemical principles. In The Pharmaceutical
making them isotonic and, thus, safe and acceptable. Sciences; Lea & Febiger: Philadelphia, 1993; 101, 142,
169–189.
The techniques of osmometry have been refined, and 10. Deardorff, D.L. Am. J. Hosp. Pharm. 1980, 37, 504–509.
now instruments that can estimate freezing-point 11. Murty, S.R.; Kapoor, J.N.; DeLuca, P.P. Am. J. Hosp.
depression, vapor pressure, or osmotic pressure from Pharm. 1976, 33, 546–551.
12. Gatlin, L.; Kulkarni, P.; Hussain, A.; DeLuca, P.P. Am. J.
microliter quantities of samples in a few minutes are Hosp. Pharm. 1979, 36, 1357–1361.
commercially available. At the same time, very few 13. Streng, W.H.; Huber, H.E.; Carstensen, J.T. J. Pharm. Sci.
pharmacists are required to compound prescriptions 1978, 67, 384–386.
14. Huber, H.E.; Streng, W.H.; Tan, H.G.H. J. Pharm. Sci.
requiring the knowledge of the various methods of 1979, 68, 1028–1032.
adjustments of tonicity. Because of ever-increasing 15. Siegel, F.P. Tonicity, Osmoticity, Osmolality and Osmo-
complexities in the structure of new drug entities, there larity. In Remington’s Pharmaceutical Sciences; Gennaro,
A.R., Chase, G.D., Marderosian, A.D., Harvey, S.C.,
is an increasing problem of their inadequate aqueous Hussar, D.A., Medwick, T., Rippie, E.G., Schwartz, J.B.,
solubility, exemplified by drugs such as Cyclosporine Zink, G.L., Eds.; Mack Printing Co.: Easton, Pennsylvania,
and Taxol. A number of organic solvents and new 1990; 1481–1498.
16. Wells, J.M. J. Am. Pharm. Assoc. Prac. Ed. 1944, 5, 99–106.
classes of surfactants are being developed and used 17. White, A.I.; Vincent, H.C. J. Am. Pharm. Assoc. Prac. Ed.
to aid in solubilization and, thus, in the formulation 1947, 8, 406–411.
of these drugs for parenteral administration. The issue 18. Sprowls, J.B. J. Am. Pharm. Assoc., Prac. Ed. 1949, 10,
348–352.
of tonicity needs to be addressed from this perspective
because organic solvents and the surfactants behave
differently in solution than do the traditional solutes
whose characteristics in solution are well-understood.
BIBLIOGRAPHY
Also, dispersed systems such as nanocapsules,
liposomes, and microemulsions are being developed Flynn, G.L. J. Parenteral Drug Assoc. 1979, 33, 292–314.
as parenteral formulations. The colligative properties Hadzija, B.W. Am. J. Pharm. Ed. 1995, 59, 191–195.
 Tooling for Tableting
Annette Bauer-Brandl
Institute of Pharmacy, Department of Pharmaceutics and Biopharmaceutics,
The University of Tromsø, Tromsø, Norway

INTRODUCTION or model does not constitute any value judgment.

Comprehensive recommended publications are ‘‘The
Introducing high-speed tableting in 24-hr shifts has Tablet Specification Manual’’[1] with respect to tech-
tremendously increased productivity, which in turn places nical and practical problems of tooling, and for all
heavy demands not only on the tablet presses and the aspects of tableting including development of tableting
compressibility of the tableting materials but also on pre- materials, optimization of compression cycles, and
cision with respect to dimensions and the wear resistance handling of tooling ‘‘Die Tablette.’’[2]
of the toolings (punch and die sets). Furthermore, in many
cases of high tableting-speeds, adhesiveness of tablets to
the toolings becomes a problem in the course of long-term
production. Tableting materials need to be optimized in GENERAL TERMINOLOGY OF TOOLING
line with new demands. At the same time, tooling require-
Technology–

ments with respect to adhesion-reducing finish have A set of tooling consists of upper punch, lower punch,
Tooling

become more important than ever. and die. The upper punch has a shorter stem; the lower
Special attention must be paid to handling and punch stem is longer because it travels longer distances
maintaining the tool sets during their entire lifetime. up and down in the die for filling, compression, and
Not only does timely mechanical work-over prolong ejection, thereby sealing the die hole from below during
their total lifetime, but also trace patterns of wear on the entire process of compression.
the tooling itself can be indicative of numerous differ- In single-punch tablet presses (reciprocating tablet
ent events during production, inter alia pressures that presses), the punches are fixed to the punch holders
are generally too high, inadequate lubrication resulting which travel up and down, and in many cases fixation
in excessive friction, blockage of pressure rollers, tilted screws are used. Each make has its own holder design
punches, or dies. Furthermore, worn-out punches not for punches and dies, and consequently for the tooling
only affect the quality of the tablets (increased vari- sets as well. Therefore little can be said in general terms,
ation of mass, reduced surface shininess etc.), but can but it would appear that on the whole the remarks on
also cause serious problems in the process (powder loss the rotary press tooling apply correspondingly.
with dust sticking to lubrication oil as well as excessive In rotary tablet presses, only the dies are fixed in the
wear of the tablet press itself). Scratched surfaces can die table, whereas the punches are vertically mobile
lead to grave problems with tablets sticking to the and are not fixed to a holder. They slide up and down
punches at the discharge point, making tablet pro- in the turret bores, driven by cam tracks guiding the
duction impossible. A deviation in the length of a punch heads. The dwell time, i.e., the period during
lower punch translates directly into mass variability which the tablet is under full compression, is deter-
in the tablet batch; variation in the length of the entire mined by the diameter of the flat part of the punch head.
set also leads to varying compression forces and conse- The amount of clearance (space) between interact-
quently affects the dissolution properties of the tablets. ing parts like punch/die and barrel/turret guide,
On the other hand, tablet tooling design opens up depends on the tolerance range (deviation from theo-
for many possibilities for overcoming common tablet- retical dimensions due to practical manufacturing
ing problems that cannot be solved by modifying the reasons) of tooling dimensions. The clearance between
materials or the compression process, such as lubri- the die wall and the punch tip affects the tableting pro-
cation problems which in general are also a function cess as well as the mechanical properties of the finished
of tooling size and shape. product: while the powder is compressed, air needs to
be released, and when the compressed tablet is moved
upwards within the die bore, friction on the die walls is
BACKGROUND affected by the clearance.
A deviation in the overall length or the working
In the following text, only examples and principles will length of a lower punch in a set of punches for a rotary
be discussed. Mentioning or omitting a specific make press translates directly into mass variability in the
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-120012014
3782 Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved.
Tooling for Tableting 3783

tablet batch; a deviation of not more than
0.1 mm Upper Head flat Outside
can lead to a mass variability of 2% in the case of small Punch head angle
tablets. Moreover, variability of the lengths of both
lower and upper punches affects the variability of com- Inside
head angle
pression forces with consequences for tablet dimen-
sions, their tensile strength, and dissolution behavior.
In the case of a short lower and upper punch, over-
filling would not necessarily lead to an increased force Key
signal.[3] Overall Working
length length

TOOLING FOR SINGLE-PUNCH PRESSES

AND COMPACTION SIMULATORS

Nowadays, single-punch presses are, in general, only Barrel to stem

chamfer
used in R&D, but hardly in pharmaceutical pro-
duction. Useful research tools are instrumented special
Cup
hydraulic presses, ‘‘compaction simulators,’’ which
allow for online measurements of forces and displace-
Die Die bore
ments during the tableting process. Compaction simu- Protection Chamfer
lators are in many cases designed to accommodate radius

Technology–
standard single-punch tooling, which is specific for Height/

Tooling
Die
each machine type, e.g., Korsch EK0 (http://www. Depth
groove
korsch.de), Kilian SP300 (http://ourworld.compuserve.
com/homepages/Kilian). Dimensions of the tooling Outside
and tolerances thereof are up to the press and tooling diameter
manufacturer.
Two-piece configuration—i.e., a punch with inter- Lower
changeable punch tips—is common for single-punch Punch Tip straight
presses as, in general, single-punch tooling is less Stem
exposed to abrasion and—compared to tooling for Barrel to stem
rotary presses—can relatively be aligned easily. In the radius
case of two-piece tools, the lower punch tips are not
rigidly fixed but allow for some tilting in order to avoid
excess friction in the die bore. Multiple tools comprise
several punch tips on a single holder in order to Barrel
increase productivity. They have been common for
single-punch presses used in small-scale production,
particularly in the form of multipiece tools, where
Barrel to neck
any cracked tip can be exchanged separately. radius
Neck
Barrel to stem
Head radius
TOOLING FOR ROTARY TABLET PRESSES

Fig. 1 shows the general terminology of punches and Fig. 1 General terminology of tooling (sets of upper and
lower punches and dies) for rotary tablet presses.
dies for rotary tablet presses. The punch consists of
head, neck, barrel, and stem. The barrel diameter and
the overall length are determined by the machine turret
dimensions. The working length (i.e., overall length International Standards
minus any cup depth at the punch tip) determines
thickness and mass of the tablets and needs to be Standardization of tooling dimensions has been agreed
controlled regularly. The variation in working length to facilitate interchangeability between presses and
should be within
10 mm to ensure low variability of to reduce the number of spare parts. In many cases, a
tablet properties. The die depth should also be accu- certain press is available with different turrets in order
rately cut in order to guarantee a flush fitting with to standardize tooling within a tablet manufacturing
the die table. company irrespective of the make of tablet presses used
 3784 Tooling for Tableting

in R&D and production, respectively. A special turret

for R&D on which all standard tooling can be used on
one and the same turret is available from Pharmachine
India (http://www.serenader.com/pharmachine/develop.
asp). It allows to interchange punches and dies of any
sizes for trial compression.
There are two main standards: 1) B-tools with sub-
categories EU19 (Europe)/TSM 19 (USA)/Japan
Norm; 2) D-Tooling, also called EU1, TSM 1, which
are classified according to the dimensions of the barrel
diameter, overall length, and the overall diameter
of the punch head. Fig. 2 compares dimensions of
B- and D-tooling. D-tooling is thicker than B-tooling;
European toolings (both B- and D-types) are longer
compared to TSM types. Fig. 3 explains terminology
of the punch head in order to differentiate subcate-
gories of B- and D-toolings. The head angle or head
radius (in the case of ‘‘domed heads’’), respectively,
determines the lead under the compression roller.
The head flat determines the dwell time of the com-
pression event (i.e., the period during which the tablet
Technology–

is under maximum pressure), which should preferably

Tooling

be long so as to yield mechanically resistant tablets.

The head flat must in all cases be smaller in diameter
than the neck in order to guarantee pressure trans-
mission onto the whole punch. Neck shape and neck
diameter are determined by the dimensions of the

Fig. 3 Punch head terminology.

cam track. The head thickness needs to be precisely

cut in order to ensure a smooth gliding of the punches
through the cam track and, in the case of lower
punches, to guarantee a reproducible dosing of granule
in the die as well. Three subcategories of B-tooling
according to the European and the US (TSM) stan-
dard as well as the Japanese norm are compared in
Figs. 4A and B in general terms, and their respective
head dimensions are accurately depicted and closely
compared in Fig. 5A. The respective head dimensions
for the three subcategories of D-tooling are shown in
Fig. 5B.
There is another head shape to the D-tooling called
Fette EU1 441 (http://www.fette.de), which is charac-
terized by the absence of a neck and a larger head
diameter (38.00 mm) allowing for increased dwell times
(see ‘‘Special Tooling’’).
A further subcategory of B-tooling, which is
referred to as B2-type (Stokes, http://www.stokesdti.
com), has the same barrel diameter as the standard
Fig. 2 Standard sizes of B- and D-tooling. Comparison of B-type, but the overall length of the lower punch is
the respective subcategories according to the European smaller (Fig. 6). B2 is used on some older presses,
standard and the U.S. standard (TSM). but not manufactured any more. There is yet another
Tooling for Tableting 3785

inversion of the dies. Tapered dies not only let air

escape, but also assist ejection as the stress is gradually
released by ejecting to an increasing diameter thereby
preventing capping and generally increasing the tensile
strength of the tablets. Furthermore, their use also
allows higher machine speeds.
Shaped punches—all other shapes than round
ones—and multitip punches need to have keys on their
barrel in order to prevent any rotation. The termi-
nology of keys is explained in Fig. 10. Keys are available
in two configurations: the Woodroff key and the flat
key (European key) can be distinguished by the shape
of the slot milled into the barrel where the key is in
many cases screwed into position. The vertical position
of the key on the punch barrel and the length of the
key itself are determined by the turret dimensions in
such a way as to allow for clearance in the punch-guide
keyways. The optimum angle of the key with respect to
tablet shape is determined by the rotation direction of
the tableting machine and the design of the take-off
fence at the discharge point with respect to the punch

Technology–
tip, and therefore not standardized. Tablets hitting

Tooling
Fig. 4 Subcategories of B-tooling, defined by overall length
and head shape: (A) U.S. (TSM) standard, and Japan norm; the fence with a corner should be avoided; they should
(B) European norm (domed head). rather hit it with a flat face in order to ensure proper
discharge without fragmentation. Common configura-
tions for punch keys and their lengths can be found in
variation on standard B-type tooling, where the overall Tablet Specification Manual.[1]
diameter of the die is smaller (Fig. 7). The die diameter Barrel flutes, usually on round upper and lower
determines the maximum tablet diameter as well as the punches, are vertical sharp-edged grooves along the
maximum number of die bores in the case of multiple length of the punch barrel. During production, they
tools. A larger die is more resistant although pressure would rotate and thereby scrape off possible deposi-
is not directly applied to it but only by lateral exten- tions of material in the punch guideways.
sion. The terminology of standard dies is explained in Several types of dust caps and sealings fitted to the
Fig. 8A. The height (depth) of the die ensures a flush barrel-to-stem chamfer (e.g., Ref.[4]) are available to
fitting with the die table. The overall die diameter— prevent contamination of the tablets by machine oil
nominal size and manufacturing tolerances—enables dripping from the upper punch. Alternatively, punch
a tight fit in the turret avoiding lateral movement, bellows in silicone rubber fitted into special seal grooves
which is also supported by the die groove accommo- for Euro-standard are available (e.g., from Holland,
dating the locking screws. A protection radius or pro- http://www.iholland.co.uk).
tection shoulder prevents damage to the die locking
screw or scoring of the die pockets in the turret caused
by burrs or sharp edges on the die. The die bore should SPECIAL TOOLING
be made as close to its nominal size as possible because
it is this, which determines the tablet size, not the Some makes of tablet presses have their own tooling
punches. A bore chamfer provides a lead for the upper specifications allowing for longer dwell times and
punch to enter the die. higher compression forces, such as the Fette EU1 441
Any clearance between the die wall and the punch mentioned earlier, and Kilian 25/32, with subtypes in
affects the tableting behavior, as entrapped air will die shape, i.e., grooved and slotted, which are depicted
escape through the gap when pressure is applied. together with standard types in a tooling identification
Fig. 9 presents the terminology. Table 1 summarizes chart (Fig. 11). Most tablet presses from the manufac-
manufacturing tolerances of punches and dies as well turers cited are alternatively available with turrets for
as the clearance between them. standard size tooling.
Die bore tapers with a conical opening as is depicted Two-piece configurations with interchangeable
in Fig. 8B have the same effect as the clearance. Tapered punch tips can also be used in punches for rotary
dies are more expensive than standard dies. Tapers can presses. Special care and experience are necessary as
be applied to both ends of the bore to allow for an these punches can be difficult to assemble and align;
 3786 Tooling for Tableting
Technology–
Tooling

Fig. 5 Head profiles of standard tooling. (A) Heads of B-tooling with subcategories European, TSM, and Japan norm. (B) Heads
of D-tooling with subcategories European, TSM, and Japan norm.

they are mostly used with multiple tools in order to punch head of shaped tooling to rotate in order to
increase productivity, thus offsetting additional costs. increase the lifetime of punches and cams.
Korsch (www.korsch.de) has a special machine for By IMA (http://www.ima.it), a special high-speed
solving sticking problems, where the entire punches machine design has been developed. Feeding happens
are rotated before and after the compression phase in at the center, taking advantage of increasing centrifu-
order to prevent adhesion. These punches have a gal forces. This design requires special tooling, which
cogwheel around their neck. is made in two pieces. This means that only the punch
From Manesty (http://www.manesty.com) a special tips need to be changed, which saves time in this special
rotating-head tooling is available which enables the machine.[5]

Fig. 6 Subcategories of B-tooling: B2 tooling compared to Fig. 7 Subcategories of B-tooling: BB tooling die diameter
standard B. compared to standard B.
Tooling for Tableting 3787

Fig. 9 Terminology of tolerances and clearances between

punches and dies.

Technology–
Tooling
In order to avoid sticking, it is recommended to
avoid corners. Compared to the profile of the land/cup
connection, an alteration of the common connection
Fig. 8 Die terminology: (A) Standard die. (B) Tapered die. between the bevel and the flat by rounding the beveled
edge of flat-faced punches, thus reducing the possibility
of sticking of material to the sharp edge, has been pro-
Instrumented punches and dies, which enable online
posed to reduce sticking problems.[6]
measurement of forces on the punch and relative dis-
Break lines are most often bisects or quadrisects,
placement or acceleration, electrical resistance, and
whereas engravings often comprise characters and
acoustic release as well as measurement of die wall
graphic objects facilitating identification of the tablets.
forces, are special tools used exclusively in research;
Engravings on tablets can be embossed or debossed
they are discussed in a separate chapter.
(Fig. 17), the punch has to be the respective opposite,
and the characters and figures are mirrored. In many
cases, break lines and engravings trigger sticking pro-
PUNCH TIPS blems. In order to quantify the pull-off force of a tablet
sticking to a punch surface, a special instrumentation
First of all, one has to make a general distinction was developed[7] and used for the optimization of the
between round and shaped tablets. The nomenclature angle between the engravings and the surface.[8] As
of tablet shapes of round flat and convex tablets as well sticking in most cases starts at the lateral faces of the
as the respective punch tip forms are shown in Fig. 12. engravings due to shear forces occurring there during
The tip edge design of the punch and the forces it compression, the authors quoted above generally
is exposed to during compression, and the resulting recommend a small angle of the stroke to the surface
deflection forces are shown in Fig. 13. It is impossible (between 45 and 75 —as to the question of how to
to maintain an infinite sharp edge as this would define the opening angle, see Fig. 18). This is in
wear off immediately; the figure also shows how a land contrast to findings of Sabir, Evans, and Jain,[9] who
is introduced by polishing the punch tip in order to recommend an angle of 90 , but their study lacks
increase its pressure tolerance. The land/cup connec- conclusiveness as the steep angle tooling, unlike the
tion can be designed as a sharp corner, a standard reference tools, was chromium-plated. Waimer et al.
curvature (radius), or a blend radius. used a cone for their measurements,[7,8] and it can be
Round tablet profile terminology can be found in assumed that engravings comprising a greater number
Fig. 14. of lateral faces per area would need even more attention.
Similarly, for shaped tablets, terminology is Empirically, it is recommended to avoid sharp corners
explained in Fig. 15, and well-known examples of as in characters like ‘‘A’’ and to use radii instead; tooling
shapes and profiles are depicted in Fig. 16. manufacturers have their special rounded character type
 3788 Tooling for Tableting

Table 1 Tolerances and clearances in standardized tooling Requirements for break lines have been increased
Tolerances for B- and D-type round tooling: during the past years. All break lines must ensure that
Upper punch Overall length
50 mm the tablet breaks into equal parts and lines for decora-
Working length
10 mm tion purposes are not legal any more. Guaranteeing a
Head diameter þ0 mm–100 mm reproducible crack is not trivial as the powder located
Lower punch Overall length
50 mm right under the break line is exposed to the highest
Working length
10 mm compression stress and most compaction and is there-
Head diameter þ0 mm–100 mm fore in many cases mechanically strongest. Therefore,
Die Height
10 mm break lines should not be too deep, and notching of
Diameter þ0 mm–10 mm the outer tablet wall serves to ensure a defined crack
Concentricity 25 mm line. Notching requires coves in the die wall and, conse-
quently, punch rotation has to be prevented. Some
Round tooling Shaped tooling typical standard bisect line types are shown in Fig. 19.
(mm) (mm) The shape, angle, and depth of bisects with regard to
the shaped tablet surface has been optimized in order
Tolerances and clearances for round and shaped tooling
to facilitate the breaking of tablets by pressing them
Nominal die bore size þ10–0 þ20–0 against a hard pad with one finger (Fig. 19, lower line,
Upper punch tip 40 60 and Figs. 20 and 21).
tolerance In bigger tablets other than those commonly used
Lower punch tip 20 40 pharmaceutically, the requirements for reproducibility
tolerance are lower, which, e.g., makes it possible to press a
Technology–

Upper punch/die 40 40 trough into the tablet surface so that a second mixture
Tooling

minimum clearance of tensides for dish-washing can be applied after tablet-

Upper punch/die 60 80 ing. The punch can be designed as an interchangeable
maximum clearance cone made from synthetic material.[10]
Lower punch/die 20 20 Multiple tools have several punch tips on one and
minimum clearance the same barrel; the corresponding die has as many
Lower punch/die 40 60 bores in order to increase productivity. One disadvan-
maximum clearance tage results from the more delicate handling of the
tools, but multiple tools are definitely useful for small
tablets in order to increase productivity. Minitablets
of 2 mm diameter are produced as a substitute for
sets (Fig. 18). For film coating, the profile of the engrav- pellets and offer the advantage of better reproducibility
ings must be particularly flat-angled in order to of filling into capsules as well as better reproducibility
ensure wetting with the coating material. The maximum of their specific surface area, which is important for
area on the surface of the tablet that can be used for the coating process and the subsequent reproducible
engravings depends on the cup depth. release of the active drug. Multiple punch tips are fre-
quently manufactured as multiple tools in a several-
piece punch design. A 19-fold punch set is available
and working in standard production of minitablets.
Up to 55-fold tooling has proved to function in experi-
mental studies (http://www.ritter-pharmatechnik.de).
The setup of the tiny tooling is depicted in Fig. 22.

MATERIALS, MANUFACTURE, FINISH,

AND PRESSURE TOLERANCE

Materials and Manufacture

Steels are basically classified as carbon and alloy steels

which are defined by national and international stan-
dards, e.g., ASTM and DIN. Carbon steels contain
less than 1.65% manganese, 0.6% silicon, 0.6% copper,
plus boron and deoxidizers as well as carbon as the
Fig. 10 Keys on punches: terminology. principal additive increasing hardness. Manganese also
Tooling for Tableting 3789

Technology–
Tooling
Fig. 11 Comparison of shapes and dimensions of well-known toolings.

increases hardness, particularly the ability to be caused by HCl released from drug salt.[11] The com-
hardened by tempering, and is present in all steels. position of special steel types for tooling, their prov-
Silicon acts as a deoxidizer. Alloy steels exceed these enance, and their thermal treatment are part of the
limits or contain additional elements, e.g., chromium, trade secret of tooling manufacturers.
vanadium, tungsten, molybdenum, nickel, and cobalt. A high abrasion resistance usually corresponds to
A high-nickel content increases resistance to corrosion high hardness of a steel type, whereas its ductility

Fig. 12 Terminology of round tablets for flat-faced and convex tablets.

 3790 Tooling for Tableting
Technology–
Tooling

Fig. 14 Round tablet profiles. (Open ring tablet: From Not-

Fig. 13 Punch tip edges: wear and deformation under load, ter, A. R. German Patent DE 4342146 C1, December 10, 1993.)
effect on tablet shape and tip deflection.

which makes heat-treatment easier. More about steel

qualities and international standardization thereof is
and toughness is low and vice versa: very hard tooling
available from steel manufacturers or the relevant
may fracture, ductile material may wear off quickly.
authorities via the World Wide Web (e.g., http://
Hardness and ductility do not only depend on the
www.sz-metal.si).
chemical composition of the steel but also on its
heat-treatment (tempering). A second tempering pro-
cedure for punch tips, which is computer-controlled Finish
and carried out at high temperatures in a vacuum fur-
nace, softens the material and prevents fracture. Adhesion, sticking, picking, and filming on punches
Punches usually have a longer lifetime than dies if and dies can in many cases be avoided by a suitable
they are made from the same material; that is why surface finish of the tooling. In some cases, the surface
manufacturers often choose different steel qualities. is smoothened during continuous use of punches,
The surface of a die bore can become smoother or whereas in other cases, depending on the material
rougher during its use. The ejection force does not— compressed, e.g., in the case of crystalline lactose, small
apart from the composition of the tableting material scratches may occur on the punch tips after a few thou-
and the compaction force—necessarily depend on sand compression events.[13] On those small scratches
roughness of the die wall, but to a high degree depends and defects in the surface, corrosion will start.
on the metal type of the die.[12] Tolerances for the surface finish of tooling are also
For specific tableting tasks, the tooling manufac- specified by manufacturers and in international stan-
turer will be able to provide optimum tool properties. dards, see Fig. 23. Any roughness of the finishing is
It may be helpful in this respect to supply him with a below 1 mm peak-to-valley height, and particularly
sample of the tableting material in question. punch-tip faces and die bore walls are polished to
Table 2 gives hardness characteristics of some steel achieve lower tolerances.
qualities that are frequently used. 440C has the highest
chromium content (17%), which increases wear resis-
tance. D2 and D3 have a high-chromium content Coatings and Platings on Punch Tips
(12%) and are additionally carbon-rich (over 1.5%).
Chromium and carbon together form stable carbide, Reduced ejection forces can help to avoid both lami-
which increases hardness. S1 contains tungsten (2%), nation and capping and give tablets a shiny surface
Tooling for Tableting 3791

Technology–
Tooling
Fig. 15 Terminology of shaped tablets: profiles of capsule-
shaped (oblong) and oval tablets. Fig. 16 Examples of well-known tablet shapes. (Courtesy of
Ritter Pharmatechnik, D-Hamburg.)

as well as high mechanical strength. Internal lubri-

cation (lubricants added to the tableting material) is affects clearances and tolerances. As this is also a part
counter-productive not only because its admixture is of the trade secret of manufacturers and specialized
another critical step in production, but also because companies, little is published on this subject.
the most effective lubricants are hydrophobous and Hard chromium plating is applied galvanically with
make the tablets softer and increase disintegration time a thickness of approximately 30 mm. It should not be
as well as dissolution rate. Attempts to coat the tooling confused with blank chromium plating, which consists
with fluid or solid lubricants just before each com- of several layers of other metals with a very thin chro-
pression event are relatively complicated with respect mium layer on top and is not suitable for tooling. Hard
to machinery, reproducibility, dust formation, and chromium platings, although appearing shiny and
GMP demands, e.g., batch definition. In many cases, smooth, are more porous than the blank coatings, brit-
an easier alternative may be to apply a finish to the tle, and in many cases not sufficiently wear-resistant.
punches that reduces friction by special coating or Moreover, any transition of traces of chromium into
plating of the punch tips so that ejection forces are the product would be totally unacceptable for pharma-
reduced. Another reason for coating the punches ceutical use. Not in all cases do they have the desired
may be to decrease the adhesiveness of tablets to the effect with respect to sticking.[14]
punches as this does not necessarily decrease with Nodular thin dense chromium is an FDA-approved
reduced ejection forces. It may also be necessary to coating which is applied to the entire punch and all its
increase corrosion resistance in cases where aggressive faces.
chemicals are compressed. A critical demand on all Chromium can be implanted into the surface in
coatings and platings is their wear resistance. The the form of chromium nitride at a depth of up
thickness of the coat must be taken into account as it to 1 mm by nitrogen ion bombardment of the surface.
 3792 Tooling for Tableting

Fig. 17 Embossing/debossing of engravings on tablets and

punches.
Technology–

This treatment proved to have a favorable effect on

Tooling

sticking problems[14] and makes the surface relatively

wear-resistant. Titanium can be applied in the same
way in the form of TiN. This treatment leaves the tools
in a ‘‘bronzed’’ appearance because the surface is

2.50

Fig. 19 Forms of commonly used bisects. (Courtesy of

0.5 Ritter Pharmatechnik, D-Hamburg.)
Angle of engraving
usually between
60fl - 100fl (normally relatively rough as seen in the electron microscope.
used to define Increased hardness and greater wear and corrosion
the angle) Angle of
75fl
resistance is achieved, but the effect on friction is smal-
engraving
relative to ler than in the case of CrN.
surface (for Nickel–chromium–boron alloying coat applied on
0.5 definition too) both punches and dies has been described[15,16] and
52.5fl
compared to untreated tools. Surprisingly, depending
Tablet
surface on the material under compression, the coating had
R0.1 the expected effect, or there was a total lack of effect,
0.3
or they even increased ejection forces. The authors
R0.1
concluded that anyway, the tablets were superior in
appearance.
Cut through engraving "A"
Titanium carbide is by far the hardest material
Fig. 18 Terminology of engravings, exemplified by the available for the treatment of surfaces.
character ‘‘A.’’ (Courtesy of Ritter Pharmatechnik, Teflon coating has not proven to be wear-resistant
D-Hamburg.) enough to tolerate tableting.
Tooling for Tableting 3793

Maximum Pressure Tolerance

The pressure tolerance depends on the smallest diam-

eter of the punch, e.g., the backing-off lathe at the
punch tip and the cup depth. It is calculated from
the load capacity of the tool steel. A guide for
maximum pressure tolerance is given in Table 3.

Lifetime

Fatigue failure is cumulative because any over-stress

inflicts microscopic damage. It correlates with the
maximum stress in each loading cycle: a 25% reduction
Fig. 20 Example of a pressure-sensitive quadrisect tablet. in stress may prolong the lifetime tenfold. It is advis-
(Courtesy of Ritter Pharmatechnik, D-Hamburg.) able not to exceed the stress level over the maximum
allowable limit for infinite lifetime; it is much better
to change the design of the punch-tip cup in order to
Carbon in layer thickness of 0.1 mm–10 mm, doped eliminate fatigue failure.
with hydrogen in 5 at.%–30 at.% has proven to be Average lifetime is most affected by granule com-
extremely wear-resistant[17] and to prevent sticking of position and lubrication properties, lying between
200,000 and several millions of compression events.[18]

Technology–
tablets. The amorphous carbon layer is applied by

Tooling
CVP (chemical vapor deposition), which can also be Principal reasons for termination of punch life are
used to apply coatings of metal, alloy, carbide, and damage to heads and tips, rolled-in or burred tip edges,
nitride. It generally yields lower adhesion and superior pitted faces, distorted and/or flatted bisects or engrav-
wear resistance of the coatings, but the high tempera- ings, and undersized tip lands. For dies, it is the wear in
tures necessary for the process (1000 C) would soften the die bore that terminates lifetime, whereas for car-
the steel and distort it. Ion plating is an alternative bide lined dies it is often distortion leading to die screw
ensuring low adhesion and minimum distortion of groove. Increased clearances accelerate wear, e.g., by
the punches. extrusion of powder between punch and die. During
lifetime the deflection of the upper punch in the turret
can double, which leads to rolled-in punch tips. Cam
Alternative Materials design and cam track tolerances determine wear on
punch heads with smaller tolerances used on modern
Inserted (lined) dies are fitted with an insert (liner) in machines prolonging punch life.
the outer shell of the metal and consist of a
much harder material such as tungsten carbide or a
ceramic. Liners are used for compression of abrasive
or corrosive materials. The outer shell of steel protects MAINTENANCE OF PUNCH SETS
the harder, more brittle liner from possible failure, e.g.,
by the die locking screw. Sets of toolings, i.e., upper punch, lower punch,
and die, should not be interchanged. Therefore, they
are usually marked by engraved numbers, punches at
the neck, dies on the outer wall. Special care must be
taken when toolings are fitted to the tablet press.
Fitting instructions usually follow with the manual
for the respective press. In general, careful aligning is
crucial, for hands-on hints see[19] and ask the tooling
manufacturer. Special maintenance kits useful for
changing punches and containing a set of aids are
available from press manufacturers and tooling manu-
facturers.
Immediately after the end of the production cam-
paign, and in the case of long-lasting production cycles
also during runs (e.g., daily), tooling should be care-
Fig. 21 Method for dividing pressure-sensitive tablets into fully cleaned. Unless a special washing machine is used,
equal parts. warm soap water and a soft brush are suitable for
 3794 Tooling for Tableting

Fig. 22 Setup of multitools, left: minitablets,

Technology–

2 mm diameter; right: triple-tool for 6-mm

Tooling

tablets. (Courtesy of Ritter Pharmatechnik,

D-Hamburg.)

thorough cleaning; if necessary, a few minutes in an one can use cost-effective ‘‘drag finishing’’ machines
ultra-sound bath can be helpful. Be careful not to (e.g., from Otec, http://www.otec.de), which drag
scratch the surfaces! Upon rinsing with warm water, and rotate punches and dies through a granulate
ethanol or isopropanol is used to remove any remain- consisting of slightly abrasive particles coated with a
ing oil. Removing punches and dies for washing takes special paraffin base. This process takes about 1 hr,
a lot of time. Some tablet presses (e.g., from Fette, depending on the abrasiveness of the granulate. The
Korsch) are constructed in a way that enables inter- process does not round off the tooling but only
changeability of the whole turret including tooling, decreases surface roughness and can be used whenever
and the cleaning of the turret takes place in a fully tooling appears dull or discolored. Furthermore, the
automatic washing machine at the same time as the punch barrels are polished too, which may help to pre-
rest of the tablet press is cleaned in place, which very vent rough motion of the punches.
much reduces turn-over-time, especially if two separate The wear pattern on punch heads, punch tips, and
turrets are available. in die bores may indicate specific problems during
Tooling should not be touched with bare fingers tableting. Measuring dimensions of punches and dies
after washing, and usually all the tooling is thoroughly is a task which has to be performed frequently and
coated with an acid-free oil in order to prevent cor- can be carried out computer-aided. Dimensions to be
rosion. Instead of expensive commercial products, checked are overall length, cup depth, working length,
paraffin and vaseline in pharmacopoeial quality are diameters, tip diameter, and die bore dimensions. Use a
also suitable. Some users want to avoid all oiling as micrometer.
oil may contribute to sticking problems; in this case For estimating the clearance of the upper punch in
punches are washed with alcohol, possibly dusted with the turret and wear of the turret guide, it is easiest to
magnesium stearate, and stored in a humidity- measure the deflection of the upper punch tip, which
controlled atmosphere.
Surface roughness and alteration in tooling dimen-
sions are results of use and wear. Tooling surfaces Table 2 Hardnesses of steel qualities used for tooling
should be frequently checked (magnifying glass). (Rockwell hardness)
Timely polishing will prolong the lifetime of the tool-
Steel S1(BS1) S7 BD2 (D2) BD3(D3) 440C
ing. Polishing can be done conventionally by hand stainless
on a lathe using ultra-fine polishing paste, polishing
Rc-hardness 54–56 54–58 58–60 58–60 56–58
wool, and felt or a polishing brass brush. Alternatively,
Tooling for Tableting 3795

0.40 0.05 clashing, dust, and humidity. Cabinets where punches

0.20 lie or hang are available in many models made from
different materials and with different degree of air-
0.20 0.40 tightness.
0.40 A special tooling utility case system, consisting of
individual punch and die trays enabling the combi-
0.20 nation of as many trays as needed, has an open design
for easy inspection and cleaning,[20] (available from
Surface finish
Holland, http://www.iholland.co.uk).
Symbol Method
Demands from GMP and the necessity of connect-
0.20 Drill/Turn
Mill ing tool sets to tablet products led to the introduction
of ‘‘tooling management’’ with the help of special soft-
Ground/Honed
ware (e.g., TM II from Natoli, http://natoli.com/, and
Lapped/Polished Vali-Scan from Holland), where information on each
Superfinish individual encoded punch and die set is collected to
0.025 provide a tooling life record. The software may be
0.80
installed on a mobile trolley and the code on each indi-
vidual tool will be checked while it is set onto the
machine.
0.20

Fig. 23 Surface finish of punches; tolerances for top-to-

Technology–
valley differences (in mm).

Tooling
CONCLUSIONS

should be approximately 100 mm. If it is more than Many common tableting problems (e.g., sticking, cap-
0.3 mm, the punches and/or guides are worn out. ping, and corrosion) which cannot be overcome by
Wear is unavoidable and it alters the dimensions of optimizing the properties of the tableting material
the tooling sets with time. It is, therefore, recommend- and/or the compression cycle, can possibly be
able to use spare sets of tooling in regular turn-over in solved by changing the tooling design. Options are
order to equalize wear of all tooling sets. the form, the material and/or the finish of the respect-
Storage cabinets for tooling should be clearly laid ive toolings. Although there are software programs
out for individual punch sets and protect them from available for designing any possible punch tip shape
and for theoretically calculating tablet volumes, sur-
face areas, and other useful features (e.g., TabletCAD,
from Natoli), this is no guarantee for being able to
compress the respective tablets in practice. Moreover,
it may be difficult to imagine on the basis of a punch
design how the tablet itself would look like. For this
purpose, simulated prototypes made quickly and inex-
pensively from a special plastic material can be bought
under the trade name Elizatab (Elizabeth Carbide,
http://www.eliz.com). As tooling manufacturers have
been working with effective tablet designs for many
years, it is generally recommendable to ask their advice
whenever questions arise as to the design of tooling or
tablets.

ACKNOWLEDGMENTS

The author wants to thank Mrs. S. Ritter, Ritter

Pharma-Technik GmbH, Hamburg, Germany, for
sharing her experience in tooling manufacture for pro-
duction and research, Mr. Rod Wolstenholm, Univer-
sity of Tromsø, for doing the drawings, and Mrs.
Gudrun Hansen, Tromsø, for help with the English.
 3796 Tooling for Tableting

REFERENCES 11. Narurukar, A.N.; Purkaystha, A.R.; Sheen, P.C. Effects of

various factors on the corrosion and rusting of tooling
material used for tablet manufacturing. Drug Dev. Ind.
1. Tablet Specification Manual (TSM); American Pharmaceu-
Pharm. 1985, 11 (8), 1487–1495.
tical Association: Washington, 1995. 12. Otz, M.; Thoma, H. Effects of the metal type and the
2. Ritschel, W.; Bauer-Brandl, A. In Die Tablette, 2nd Ed.; roughness of the die wall on the expanded work for tablet
Editio Cantor Verlag: Aulendorf, Germany, 2002. ejection. Pharm. Dev. Technol. 2000, 5 (1), 19–26.
3. Ling, W.C. Tooling as a factor in tablet weight variation. 13. Müller, B.W. Über Veränderungen der Pressflächen von
J. Pharm. Sci. 1973, 62 (12), 2007–2011. Tablettierwerkzeugen. Die Pharmazeutische Industrie
4. Notter, M.; Notter, K. Tablettierwerkzeug, Insbesondere Table- 1972, 34 (12), 972–976.
ttierstempel. European Patent 0 841 152 A2, October 24, 1997. 14. Schumann, S. The effects of chromium nitride ion bom-
5. Fabbri, G.B. Tool-Holder for Tablet Making Machine. bardment treatment of tablet tooling on tablet adherence.
US Patent 5,882,696, March 16, 1999, I.M.A. Industria Drug Dev. Ind. Pharm. 1992, 18 (19), 1037–1061.
Macchine Automatiche. 15. Tsiftsoglou, T.B.; Mendes, R.W. Effect of boron alloy
6. Cargille, J.J. Tooling Face Configuration—Particularly coating of tableting tools. Pharm. Technol. 1982, 6 (April),
Adapted for Forming Tablets (Cargille Curve). US Patent 30–32, 36–38.
5,164,206, November 17, 1992. 16. Shah, K.B.; Augsburger, L.L.; Shangraw, R.F. Effect of
7. Waimer, F.; Krumme, M.; Danz, P.; Tenter, U.; Schmidt, boron alloy coating of tableting tools. Pharm. Technol.
P.C. A novel method for the detection of sticking of tablets. 1982, 6 (April), 31–34, 41–42, 44, 49–50, 52, 54.
Pharm. Dev. Technol. 1999, 4 (3), 359–367. 17. Hieke, A.; Grischke, M.; Brand, J. Vorrichtung zum Ver-
8. Waimer, F.; Krumme, M.; Danz, P.; Tenter, U.; Schmidt, pressen von Fliessförmigen Feststoffen. German Patent
P.C. The influence of engravings on the sticking of tablets. DE 198 10 969 A1, March 13, 1998..
Investigations with an instrumented upper punch. Pharm. 18. Swartz, C.J.; Anschel, J. Evaluation of tableting tool life
Dev. Technol. 1999, 4 (3), 369–375. records. J. Pharm. Sci. 1968, 57 (10), 1779–1782.
9. Sabir, A.; Evans, B.; Jain, S. Formulation and process opti- 19. Ritschel, W.; Bauer-Brandl, A. Tablettierwerkzeuge. In Die
mization to eliminate picking from market image tablets. Tablette, 2nd Ed.; Editio Cantor Verlag: Aulendorf,
Int. J. Pharm. 2001, 215 (1–2), 123–135. Germany, 2002; 552–567.
10. Holderbaum, T. Tablettierstempel. German Patent DE 199 20. Bal, G.W. Tooling Utility System. US Patent 6,112,896,
Technology–

08 027 C1, February 2, 1999, Henkel KGaA. September 5, 2000.

Tooling
Trace Level Impurity Analysis
Daniel L. Norwood
Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, Connecticut, U.S.A.

Fenghe Qiu
James O. Mullis
Department of Analytical Sciences, Boehringer Ingelheim Pharmaceuticals, Inc.,
Ridgefield, Connecticut, U.S.A.

INTRODUCTION ingredients (i.e., excipients) that actually or potentially

exist in APIs and/or final drug product at release or
The issue of trace level impurities in active pharmaceu- within the intended expiration period of the drug
tical ingredients (termed ‘‘Drug Substances’’) and product. International Conference on Harmonization
formulations developed for APIs (termed ‘‘Drug Pro- (ICH) guidances Q3A (R) (Impurities in New Drug
ducts’’) is a broad and complex one. Impurities can be Substances),[1] Q3B (R) (Impurities in New Drug Pro-
both organic and inorganic, and can be introduced either ducts),[2] and Q3C (Impurities: Guideline for Residual
to the drug substance or to the drug product from a Solvents),[3] provide classifications and definitions of
variety of sources. These sources include the synthetic various impurities in drug substance and drug product,
process for the drug substance, ingredients in the formu- as well as the reporting, identification and qualification
lation, container closure systems, and the environment. thresholds for impurities. These guidances also discuss
Modern analytical chemistry and instrumentation have analytical procedures, impurity specifications, and quali-
enabled the identification, quantitation, and control of fication of impurities. Q3A(R) classifies impurities in new
both organic and inorganic impurities at very low levels drug substance as follows:
in both drug substance and drug product. The goal of this

Trace–Ultrasonic
article is to review and discuss the current state of affairs 1. Organic impurities, which ‘‘can arise during the
in pharmaceutical development with respect to the trace manufacturing process and/or storage of the
level analysis of impurities in drug substance and drug new drug substance.’’[1] These can be identified
product. The article includes a discussion on the scope or unidentified, volatile or non-volatile, and
of the impurities issue and the regulatory considerations include starting materials, by-products, inter-
involved, followed by a survey and discussion of the ana- mediates, degradation products, reagents,
lytical/instrumental techniques now in routine use for the ligands, and catalysts.
trace level identification of organic and inorganic impuri- 2. Inorganic impurities, which can result from the
ties, with emphasis on Mass Spectrometry (MS) and synthetic process for the API. These are usually
Nuclear Magnetic Resonance Spectroscopy (NMR). known and identified, and include reagents,
An MS-based systematic approach to the identification ligands, catalysts, heavy metals [and other
of trace level organic impurities is presented, illustrated residual metals, inorganic salts, and other mate-
with examples of original data. Although emphasis is rials (e.g., filter aids, charcoal)].
placed throughout the review on scientific and technical 3. Residual solvents, which are defined as ‘‘inor-
aspects of the impurities issue, all discussion is fully ganic or organic liquids used as vehicles for
grounded in and correlated with our current understand- the preparation of solutions or suspensions in
ing of the regulatory environment and current regulatory the synthesis of a new drug substance.’’[1] It is
guidances. The article includes references to applicable interesting to note that Q3C defines residual
scientific literature, and, as stated above, original data solvents in pharmaceuticals as ‘‘organic volatile
are used to illustrate important scientific concepts. chemicals that are used or produced in the
manufacture of drug substances or excipients,
SCOPE OF THE IMPURITIES ISSUE or in the preparation of drug products.’’[3]
Residual solvents are classified by a risk assess-
Classification of Impurities in Drug Substance ment approach into three classes:[3]
and Drug Product

Class 1 solvents: Solvents to be avoided (known
Pharmaceutical impurities are those chemical entities human carcinogens, strongly suspected human
other than the APIs themselves and known formulation carcinogens, and environmental hazards).
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-120041589
Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved. 3797
 3798 Trace Level Impurity Analysis

Examples[4] benzene, 1,2-dichloroethene, 1,1- related to impurities. As an example, they cite the case
dichloroethene, 1,1,1-trichloroethane of the excipient Lecithin in which complete control

Class 2 solvents: Solvents to be limited (non- of the compositional profile is required, including
genotoxic animal carcinogens or possible causa- the profiles of phosphatidyl choline, phosphatidyl
tive agents of other irreversible toxicity such as ethanolamine, phosphatidyl inositol, lysophosphatidyl
neurotoxicity or teratogenicity, solvents suspected choline, phosphatidic acid, triglycerides, fatty acids,
of other significant but reversible toxicities). and carbohydrates.[6,8] In a complex natural product
Examples:[4] acetonitrile, cyclohexane, and mixture like Lecithin, it is not clear exactly which trace
methanol level chemical entities would be considered as impuri-

Class 3 solvents: Solvents with low toxic ties, and which as normal components of the excipient
potential (solvents with low toxic potential to mixture. They also cite the case of Compendial pro-
man; no health-based exposure limit is needed). pellants, in which assay and control for 22 individually
Examples:[4] acetone, ethanol, and ethyl ether named trace level impurities is recommended, along
with assay and control of total chloromethanes, and
total unspecified impurities.[6,8]
Q3B(R) addresses ‘‘only those impurities in new
Although extractables and leachables are not
drug products classified as degradation products of
addressed by the available impurity guidance docu-
the drug substance or reaction products of the drug
ments, their characterization and control is clearly
substance with an excipient and/or immediate con-
important for certain drug product types, especially
tainer closure system.’’[2] These are collectively referred
for OINDP.[7,8] The United States Food and Drug
to as degradation products in the guidance.[2] It is
Administration (USFDA) has classified drug product
interesting to observe that Q3A(R) and Q3B(R)
and dosage form types with respect to risk of interac-
specifically exclude the following:
tion with the container closure system, and therefore
the likelihood of extractables and leachables being a
1. Extraneous contaminants that should not occur pharmaceutical development issue.[9] The drug product
in new drug substances and are more appropri- types of greatest concern, and therefore requiring the
ately addressed as good manufacturing practice greatest degree of extractables/leachables characteriza-
Trace–Ultrasonic

(GMP) issues. tion and control, are OINDP of all types, injectables,
2. Enantiomeric impurities. ophthalmics, and transdermals. At present, there are
no guidelines as to identification, reporting, and
Q3B(R) also excludes ‘‘impurities arising from qualification thresholds for extractables and leach-
excipients present in a new drug product or extracted ables. A comprehensive discussion of extractables
or leached from the container closure system.’’[2] and leachables is presented in a separate entry of this
Excipient-related impurities have been discussed encyclopedia.
in detail by Erickson[5] and Norwood and Qiu.[6]
Erickson[5] addresses the regulatory environment,
identification, and control issues for excipient related Control of Impurities and Residual Solvents
impurities in general and observes:
ICH Q3A(R) and Q3B(R) provide detailed maximum
‘‘Presently, there are no guidelines that are specifically daily intake (MDI)-based reporting, identification,
directed at the identification (definition) and control of and qualification thresholds for organic impurities in
impurities in excipients.’’ new drug substances and drug products. For a new
drug substance, taken from ICH Q3A(R), these
However, Norwood and Qiu[6] observe that for orally are presented in Table 1. It should be noted that the
inhaled and nasal drug products (OINDPs), FDA identification threshold is either 0.05% or 0.1%
guidance documents[7,8] require a greater degree of depending on the MDI. For ‘‘degradation products’’
excipient characterization and stricter quality control in new drug products, ICH Q3B(R) has a somewhat

Table 1 ICH Q3A(R) reporting, identification, and qualification thresholds for impurities in drug substance
MDI (dose) Reporting threshold Identification threshold Qualification threshold
2 g/day 0.05% 0.1% or 1.0 mg/day intake 0.15% or 1.0 mg/day intake
(whichever is lower) (whichever is lower)
>2 g/day 0.03% 0.05% 0.05%
(From Ref.[1].)
Trace Level Impurity Analysis 3799

Table 2 ICH Q3B(R) reporting and identification API-related organic impurities usually involves high
thresholds for impurities in drug product performance liquid chromatography (HPLC)-based
MDI (dose) Reporting threshold analytical methods, with relatively non-specific detec-
1 g/day 0.1% tion techniques such as Ultraviolet/Visible (UV/Vis)
>1 g/day 0.05% Spectrophotometry. Residual solvents analysis usually
MDI (dose) Identification threshold involves gas chromatography (GC)-based analytical
methods, again with relatively non-specific detection
<1 mg 1.0% or 5 mg TDIa
techniques such as flame ionization (FID). GC-based
1–10 mg 0.5% or 20 mg TDI
>10 mg–2 g 0.2% or 2 mg TDI methods are most appropriate for volatile analytes such
>2 g 0.10% as residual solvents, whereas HPLC-based methods are
a
Total daily intake.
more appropriate for the relatively non-volatile and
(From Ref.[2].) polar API-related analytes.
Identification of organic impurities detected and
reported at or near the required thresholds with
analytical methods based on HPLC and GC requires:
more complicated recommendation for reporting,
identification, and qualification thresholds (Table 2).
For both drug substance and drug product, the 1. A detection technique(s) capable of producing
impurity identification guidelines suggest thresholds chemical information specific to the molecular
well into the ‘‘trace analysis’’ concentration range, structure of each individual organic impurity
requiring the state of the art in analytical chemistry at sensitivities matching those of the reporting
and instrumentation. Further, both guidances suggest analytical technique/method (e.g., HPLC/UV
that lower thresholds can be appropriate if a particular and GC/FID).
impurity is determined to be unusually toxic. 2. Direct and easy interface between the organic
The level of Class 1 residual solvents should be impurity detection/reporting analytical tech-
strictly controlled below the concentration limits for nique/method (HPLC or GC) and the
every individual solvent (for example the limit for compound specific detection technique(s).

Trace–Ultrasonic
benzene is 2 ppm). Class 2 solvents are controlled
according to the permitted daily exposures (PDEs) The analytical techniques that best meet these
and maximum daily dose (Option 1 and Option 2). criteria are those based on the combination of chroma-
ICH Q3C provides PDEs for all Class 2 solvents. tography with MS, either liquid chromatography/MS
For Class 3 Solvents, ICH Q3C suggests that a PDE (LC/MS) or GC/MS. The most commonly used
of 50 mg/day or less would be acceptable without approaches for trace level organic impurity identifi-
justification. For solvents for which no adequate toxi- cation are therefore MS based. Note that an overview
cological data are found, manufacturers should supply of MS and its application in the pharmaceutical
justification for residual levels of these solvents in industry was presented in the previous edition of this
pharmaceutical products. treatise.
The guidances recommend that inorganic impurities
be detected and quantified using pharmacopeial or
other appropriate procedures, and that acceptance OVERVIEW OF MS-BASED ANALYTICAL
criteria be based on pharmacopeial standards or known TECHNIQUES
safety data.
GC/MS

Residual solvents are amenable to GC/MS analysis

IDENTIFICATION OF ORGANIC IMPURITIES because they possess sufficient volatility to enter the
AT TRACE LEVELS gas phase without thermolytic decomposition, and
they do not typically interact with the sample intro-
Analytical procedures for drug substance, drug product duction system (i.e., GC separation column, detector
organic impurities, and residual solvents are discussed interface, or detector) in such a manner as to cause
only briefly in the ICH guidance documents.[1–3] How- irreversible surface adsorption or surface catalyzed
ever, a review of the voluminous scientific literature on decomposition. The application of GC/MS for identi-
the subject, as well as our understanding of current fication of residual solvents is best illustrated with an
pharmaceutical industry practice suggests that impurity example. Fig. 1 shows a total ion chromatogram
analyses are most often accomplished by chromato- (TIC) from the headspace GC/MS analysis of a drug
graphic procedures. Detection and reporting of substance dissolved in a relatively high boiling solvent
 3800 Trace Level Impurity Analysis

x24 14.22
100
Dissolving solvent

Unknown residual
solvent

5.88

11.84

1.42

2.52

0 Time
2.50 5.00 7.50 10.00 12.50 15.00 17.50

Fig. 1 TIC from the GC/MS analysis of residual solvents in a drug substance. Note the very large and saturated peak for the
dissolving solvent, and the much smaller well resolved peaks for residual solvents.

(note that this particular drug substance is not water index) of an authentic reference compound with those
soluble). The labeled peaks in the TIC are from of the unknown.
residual solvents in the drug substance sample (the The single most important piece of information in a
Trace–Ultrasonic

very large saturated peak from the dissolving solvent mass spectrum is the molecular ion, which indicates the
should be noted). GC/MS analyses most often employ monoisotopic molecular weight of the analyte.
one of two complementary ionization processes, elec- McLafferty and Turecek[10] have discussed in detail
tron ionization (EI) or chemical ionization (CI). This the recognition of the molecular ion in an EI mass
is because both EI and CI are gas phase ionization spectrum. For GC/MS molecular ion confirmation, it
phenomena and are therefore well suited to interface is often useful to employ an alternative ionization tech-
with a separation technique (e.g., GC), which is also nique called CI. CI has been discussed and reviewed in
accomplished in the gas phase. The EI ionization pro- detail by Harrison.[12] In CI, the ion source (which is
cess is based on the interaction of an energetic electron usually a standard EI source with certain modifica-
beam (70 eV) with neutral analyte molecules in the gas tions) is pressurized to about 1 torr with a so-called
phase, producing a radical cation, or molecular ion ‘‘reagent gas,’’ which is ionized by the electron beam
(Mþ), which can undergo fragmentation in the gas producing a steady-state concentration of reagent gas
phase after redistribution of excess internal energy ions whose identity and relative concentration in the
through the chemical bonds of the molecular ion. EI ion source depend on the identity of the gas, its press-
fragmentation processes and mechanisms have been ure within the ion source, and the source temperature.
extensively studied and can be interpreted and pre- Commonly used reagent gasses include methane,
dicted based on fundamental chemical principles.[10,11] isobutane, and ammonia. The reagent gas ions in the
Further, owing to the kinetic nature of the process, source can then collide and react with neutral analyte
EI spectra are highly reproducible from one mass spec- molecules eluting from the GC column in the gas
trometer to another, making it possible to employ phase. For example, in the case of ammonia, one
computerized libraries of spectra compiled from many usually observes proton transfer producing [M þ H]þ
instruments and sources to aid in compound identifi- and cluster ions such as [M þ NH4]þ. Each reagent
cation. Fig. 2 shows an EI spectrum (A) from one of gas has its own unique suite of reacting ions, and its
the residual solvent peaks in the Fig. 1, along with a own analyte selectivity for gas phase reaction. Note
‘‘best-fit’’ library search result (B). The library search that CI is a thermodynamically controlled rather than
result suggests that the unknown residual solvent is a kinetically controlled process, which can result in sig-
butyl acetate, a Class 3 solvent.[4] This tentative nificant variability in CI spectra over time and between
library search identification would require confir- instruments. Computerized libraries of CI mass spectra
mation by matching GC retention time (or retention therefore have very limited utility.
Trace Level Impurity Analysis 3801

Fig. 2 EI mass spectra from an unknown residual solvent in a sample of drug substance (see the chromatogram in Fig. 1), and
the ‘‘best fit’’ computerized library search result (butyl acetate).

The analytical chemistry and regulatory aspects of Alsante et al.[20], Ermer and Vogel,[21] Ganguly et al.[22],
residual solvents in pharmaceutical products have been and Niessen.[23]
reviewed in detail by Witschi and Doelker[13] and Modern LC/MS systems employ two ionization
B’Hymer.[14] Both of these reviews discuss the processes that are accomplished at atmospheric press-

Trace–Ultrasonic
central role of GC-based analytical methods for this ure, termed electrospray (ESI)[24] and atmospheric
application. Also of note is the review by Dwivedi.[15] pressure CI (APCI).[25] For additional information
and references, the reader is again referred to the entry
on MS in the previous volume of this treatise. In a typi-
LC/MS cal ESI source, the eluent from the HPLC column
composed of liquid mobile phase and analyte mole-
There are many organic compound classes that owing cules passes through a stainless steel capillary with a
to poor volatility, thermal instability, or polarity are high positive or negative potential applied to the end
not amenable to analysis by GC. A great many of these (3–5 kV).[26] The electric field causes instantaneous
are amenable to separation and detection by analytical vaporization (termed nebulization) of the eluent into
methods employing LC, most often HPLC. HPLC a spray of very small droplets that pass through an
employs a liquid mobile phase with the separation evaporation chamber, which is heated slightly to pre-
column held at or slightly above room temperature. vent condensation. During this process, the droplets
Analyte volatility is therefore not relevant, and thermal evaporate rapidly, reduce in size, and increase their
decomposition is not usually possible. For a detailed surface charge density until charge repulsion causes
discussion of HPLC, the reader is referred to the treat- ions (both mobile phase and analyte) to be ejected.
ise by Snyder, Kirkland, and Glajch[16] HPLC systems The vaporization process is so rapid that equilibrium
can include various detectors, including spectrophoto- is not attained and thus many of the droplets have a
meters (single or variable wavelength UV/Vis, significant positive or negative surface charge. These
diode-array UV/visible, fluorescence) and mass spec- ions are guided into the mass spectrometer for analysis.
trometers (LC/MS). Although detectors such as diode In APCI, the ESI capillary probe is replaced by an
array can provide some compound-specific infor- externally heated probe (usually held between 300
mation, LC/MS is by far the most useful system for and 500 C) through which the HPLC column eluent
the qualitative analysis of many types of impurities passes. The heated probe, assisted by nebulization
in pharmaceutical products, including API-related gas (nitrogen), produces a fine mist of droplets, which
structures. For additional references on LC/MS and enter the region of a corona discharge. The discharge
its application to pharmaceutical product impurity ionizes mobile phase molecules producing a steady-
analysis, the reader is referred to the reviews by state population of mobile phase ions in the source.
Lim and Lord,[17] Hoke et al.[18], Krstulovic et al.[19], These mobile phase ions can undergo ion–molecule
 3802 Trace Level Impurity Analysis

A
Unknown impurities 104.48
100
m/z 722

0
B
63.53
100 x24
UV@254 nm
%

-6

C
105.85
100
API
TIC

3 Time
20.0 40.0 60.0 80.0 100.0 120.0

Fig. 3 Various chromatograms (C): TIC; (B): UV at 254 nm; (A): m/z 722 extracted ion current profile) from the positive ion
ESI LC/MS impurity profile analysis of a drug product. Note the two unknown trace level impurities in the m/z 722 extracted
ion current profile.

reactions with neutral analyte molecules to produce generation systems. The power of modern LC/MS is
Trace–Ultrasonic

analyte molecular ions, just as is accomplished in gas also best illustrated by an example. Fig. 3 shows an
phase CI (hence the term APCI). The resulting ions in-line UV chromatogram (254 nm), a TIC, and an
are then guided into the mass spectrometer for analysis extracted ion current profile (m/z 722) from the ESI
as in ESI. Note that the entire ESI and APCI processes LC/MS analysis of a drug product. ESI mass spectra
take place outside of the mass spectrometer (i.e., the from the two m/z 722 trace level impurities are shown
high vacuum region), which serves to make such LC/ in Fig. 4. Note that the [M þ H]þ at m/z 722 is con-
MS systems extremely rugged compared to previous firmed by [M þ Na]þ at m/z 744 and [M þ K]þ at

722
100

% [M+H]+
723
744
655 658 688 703 718 741 760 766 779 814
0
722
100

[M+Na]+
[M+K]+

% 723

744
760
0 m/z
640 660 680 700 720 740 760 780 800 820

Fig. 4 Positive ion ESI mass spectra from two isobaric (molecular weight 721 amu) trace level impurities (see chromatograms in
Fig. 3). Note that both impurities have [M þ H]þ at m/z 722, confirmed by [M þ Na]þ at m/z 744 and [M þ K]þ at m/z 760.
Trace Level Impurity Analysis 3803

m/z 760 (note that such adduct ions are commonly (such as nitro groups, boron, or multiple hydroxyl
used to confirm molecular weight). The reader is groups). This technique is accomplished by the addition
advised that the above descriptions of ESI and APCI of small amounts of chloroacetonitrile to the HPLC
are intended to be general, and that instrumentation mobile phase entering an APCI source. The corona
from various manufacturers can differ in detail. discharge in the APCI source produces a steady-state
Empirically, ESI and APCI have similarities and concentration of Cl ions in the gas phase which can
differences and are in many ways complementary. ‘‘attach’’ to certain analyte molecules [M] forming
Both operate at atmospheric pressure, are relatively [M þ Cl] molecular ions. Norwood and Qiu[6] have
‘‘soft’’ thermodynamically controlled ionization pro- used this technique to analyze impurities and excipients
cesses producing little molecular ion fragmentation, in nitroglycerin ointment. As an example of LC/
and are extremely rugged and robust. ESI often reflects MS ionization processes, consider the drug substance
solution chemistry with mobile phase pH therefore Nifedipine (I), which is the prototype of the dihydropyr-
being a factor, and multiply charged ions often idine class of Ca channel blockers.[31,32]
observed. Analyte molecules with high proton affinity
(e.g., amines and aromatic heterocycles) can be
expected to have reasonable positive ion sensitivity in
ESI, whereas analyte molecules with easily abstracted
protons (e.g., carboxylic acids, sulfonic acids, and phe-
nols) can be expected to have reasonable negative ion
sensitivity. ESI also performs best with most ion
sources at relatively low HPLC flows (approximately
10 mL/min), requiring flow splitting for analytical scale
HPLC. In contrast, APCI is a gas phase process with
gas phase ion chemistry in control. Even so, general
trends in APCI sensitivity are similar to those of ESI,
with high proton affinity analytes performing best in
positive ion mode and organic acids and similar chemi-

Trace–Ultrasonic
cal entities performing best in negative ion mode. Based only on the molecular structure of Nifedipine,
APCI also tends to work best in most ion sources at consider the following:
higher HPLC flows (1 ml/min, for example), making
it a good match for analytical scale HPLC. Ionization 1. The ring secondary amine group should have
processes for LC/MS continue to be developed and sufficient proton affinity to allow for positive
refined. A notable example is atmospheric pressure ion ESI and/or APCI sensitivity.
photoionization (APPI),[27,28] which is capable of ioniz- 2. There are no easily abstractable protons, and
ing certain weakly polar analytes and thus acting as an therefore negative ion sensitivity will likely be
alternative to ESI and APCI for such molecules. poor.
The APPI source employs a high-fluence gas discharge 3. The nitro group has a strong dipole, and there-
lamp, which generates vacuum-UV photons of 10 and fore chloride ion attachment is a possibility.
10.6 eV.[29] These energies are sufficient to ionize most
analyte molecules (first ionization potential 7–10 eV), Fig. 5 shows a comparison based on extracted ion
but insufficient to ionize most commonly used HPLC current profiles of ionization efficiencies for Nifedi-
mobile phase solvents (e.g., water 12.6 eV and acetoni- pine. Included are positive ion APCI and ESI, along
trile 12.2 eV).[29] The APPI process is therefore capable with negative ion chloride ion attachment APCI. As
of ionizing many types of chemical entities with low predicted, Nifedipine can be readily ionized by all three
ionization potentials without generating large back- LC/MS ionization processes. The mass spectra of
ground ions from mobile phase constituents. APPI Nifedipine obtained using these three ionization pro-
does however appear to perform better when a dopant cesses are shown in Fig. 6. Positive ion ESI and APCI
such as acetone is introduced into the APPI source both show [M þ H]þ at m/z 347 (note the
along with analyte molecules.[29] The application of [M þ Na]þ ion at m/z 369 in the ESI spectrum) with
APPI for pharmaceutical analysis has been reviewed some fragmentation (the ion at m/z 315 likely results
by Hsieh and Wang.[30] from loss of methanol from one of the methyl ester
It is also possible to assist and/or alter ionization groups). The negative ion chloride ion attachment
processes by addition of certain ingredients to the HPLC spectrum shows the anticipated [M þ Cl] at m/z
mobile phase. Negative ion chloride attachment APCI 381, along with a [M þ Cl – HCl] ion at m/z 345.
LC/MS is one such technique useful for certain weakly Note that there is also a relatively small [M þ TFA]
polar analytes with relatively strong internal dipoles (TFA) ion at m/z 459. If TFA is present in the HPLC
 3804 Trace Level Impurity Analysis

A
0.94 x10 3.27

APCI+ m/z 347

%

1
0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00

B
x24 3.34
100
ESI+ m/z 347

0
0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00

C
x36 2.77
100
ACPI–/Cl m/z 381
%

0 Time
0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00

Fig. 5 Extracted ion current profiles from LC/MS analyses of equal quantities of Nifedipine (A): m/z 347 from positive ion
APCI; (B): m/z 347 from positive ion ESI; (C): m/z 381 from negative ion chloride ion attachment APCI.

mobile phase or has ever been used on the LC/MS solvents (such as CD3OD) are added to the HPLC
Trace–Ultrasonic

system, it is possible to see such gas phase adduct ions. mobile phase for either ESI or APCI LC/MS.
Another mobile phase modification technique is Analyte molecules with exchangeable hydrogen atoms
termed Hydrogen/Deuterium (H/D) exchange,[33] in (i.e., COOH, OH, NH2, NHR, and SH) will exchange
which deuterium oxide and/or other deuterated these for deuterium, either in solution or in the gas

A
381.2
100 [M+Cl] − [M+TFA] −
APCI–w/Cl-attachment

% 383.2
345.2 384.1 459.3
222.1
0

B
[M+H] +
347.2
100
ESI+ 315.2
%
316.2 348.2
254.1 271.1 329.2 369.2 392.3 410.3
0

C
347.2 [M+H] +
100 315.2
APCI+
%
271.2 316.2 348.2
283.2
239.1 254.1 329.2
0 m/z
200 220 240 260 280 300 320 340 360 380 400 420 440 460 480

Fig. 6 Mass spectra of Nifedipine from various LC/MS experiments (A): negative ion chloride ion attachment APCI, note that
TFA is trifluoroacetate; (B): positive ion ESI; (C): positive ion APCI.
Trace Level Impurity Analysis 3805

[M + D]+ 534 536

API after H/D 538
10
exchange
% 539
540
0
535
10 API before H/D [M +H]+ 533
exchange 537
% 538
507
0
10 510 Degradant after
512
H/D exchange
%
508 513
0
505
10 507 Degradant before
% H/D exchange
508 509
0 m/z
50 50 51 51 52 52 53 53 54 54

Fig. 7 Positive ion LC/MS spectra of an API and its trace level degradation product before and after H/D exchange.

phase. Because the atomic weight of H is 1 amu and hydrogens) which suggests the presence of four
that of deuterium is 2 amu, when H is replaced by a exchangeable hydrogens:
D in a molecular or fragment ion, the m/z value for
that ion will increase by an amount directly pro-
portional to the number of exchangeable hydrogens ð1Þ ½M þ Hþ ¼ 505 therefore M ¼ 504
in the analyte molecule. As an example, Fig. 7 shows
mass spectra expanded around the molecular ion
region of an API and an associated trace level degrada- ð2Þ ½M þ D  xH þ xDþ ¼ 510
tion product before and after H/D exchange. The API ½504 þ 2  4ð1Þ þ 4ð2Þ ¼ 510

Trace–Ultrasonic
shows a [M þ H]þ at m/z 533 and a predicted therefore x ¼ 4
[M þ D]þ at m/z 534, indicating no exchangeable
hydrogens in the molecule. By contrast, the degradation
product shows an [M þ H]þ at m/z 505, and after Table 3 presents an overview of so-called ‘‘hyphe-
H/D exchange a molecular ion [M þ D  xH þ nated’’ MS techniques, with their individual suitability
xD]þ at m/z 510 (where x is the number of exchangeable and features.

Table 3 An overview of MS—hyphenated techniques

Hyphenated
technique Suitability and application Features
GC-EI/MS Small molecules Gas phase ionization Mþ.
Non-polar to medium polar (molecular ion is a radical cation)
Volatile and thermally stable Significant/predictable fragmentation
Non-reactive with analytical system (library searchable spectra) Super
separation power (capillary GC)
GC-CI/MS Same as GC-EI/MS ‘‘Soft’’ ionization process
Complement to GC-EI/MS
Molecular weight confirmation
HPLC-ESI/MS Polar, small and large molecules, Liquid phase ionization, preionized,
basic or acidic [M þ A]þ, A ¼ H, Na, K, NH4, [M  H]
‘‘Soft’’ ionization process
HPLC-APCI/MS Weakly polar to medium polar, Gas phase ionization
small to medium sized molecules [M þ H]þ, Mþ, [M  H]
With chloride ion attachment: poorly Can be used with chloride
ionizable molecules with strong net ion attachment
dipoles
 3806 Trace Level Impurity Analysis

Mass Analyzers and Accurate The ion trap and FTICR instruments are also
Mass Measurements capable of sequential MS/MS, or so-called MSn
experiments. As an example, consider the known
Different types and configurations of mass analyzers degradation product of Nifedipine, nitrosodehydroni-
can also be employed to produce additional structure fedipine (II; molecular weight 328, Ref.[31]):
elucidation information from either GC/MS or LC/
MS experiments. Fragmentation of molecular ions
can be enhanced using tandem MS (MS/MS) experi-
ments[34] with collision-induced dissociation (CID).
For example, Fig. 8 shows MS/MS product ion spec-
tra of the unknown drug product impurities shown in
Fig. 3 using a triple quadrupole mass spectrometer.
A detailed schematic and description of a triple (or
tandem) quadrupole mass spectrometer is explained
in Ref.[34]. In this experiment, the first quadrupole
selects the parent ion (in this case m/z 722 which is
the [M þ H]þ of the two trace level impurities) and
passes this ‘‘parent ion’’ to the second quadrupole, Fig. 9 shows a positive ion ESI mass spectrum from
which is pressurized with a ‘‘collision gas’’ (in this case this degradation product acquired on a quadrupole ion
argon). Collisions between the parent ions and the trap, followed by a series of CID-MS/MS spectra in
argon produce excess internal energy in the parent which the most abundant ion in the previous spectrum
ions, which can in turn induce fragmentation. The was selected as a parent (or precursor) ion. For
third quadrupole then mass analyzes the resulting frag- example, the spectrum second from the top in Fig. 9
ment (or product) ions, producing a CID mass spec- is a CID spectrum of the precursor ion m/z 329, the
trum of fragment ions that are derived from a known [M þ H]þ of II, the next spectrum down is a CID
parent. The similarities between the two impurity spectrum of the precursor ion m/z 270, and so on to
CID spectra in Fig. 8, which indicates that these two MS6. The utility of this mass spectral information,
Trace–Ultrasonic

impurities have a similar molecular structure should and its ability to establish precursor-product relation-
be noted. Other types of mass spectrometer, including ships, should be obvious.
the Quadrupole Ion Trap and Fourier-Transform Ion FTICR, magnetic sector, and time-of-flight (TOF)
Cyclotron Resonance (FTICR), and Magnetic Sector mass spectrometers are also capable of acquiring accu-
mass spectrometers, can also accomplish MS/MS rate mass measurements on molecular and fragment
experiments and are in common use for pharmaceuti- ions from both GC/MS and LC/MS experiments on
cal impurity structure elucidation. trace level impurities. Applications of FTICR LC/MS

704
100
Impurity A

104
% 122

201
133 343 347 722
159
0
704
100
104
Impurity B

%
122
201
133 159173 347 722

0 m/z
100 150 200 250 300 350 400 450 500 550 600 650 700 750

Fig. 8 MS/MS (product ion) ESI mass spectra of two isobaric trace level drug product impurities acquired on a triple quadru-
pole mass spectrometer (note that in both spectra the parent ion is the [M þ H]þ m/z 722).
Trace Level Impurity Analysis 3807

329
100

50
330
0
270
100

Products of m/z 329 254

50 284
237 268 297
0
253
Relative Abundance

100
Products of m/z 270
50

0
221
100
Products of m/z 253
50
253
193
0
193
100
Products of m/z 221
50
152
0
152
100
Products of m/z 193
50 166
193

Trace–Ultrasonic
0
100 150 200 250 300 350
m/z

Fig. 9 MSn spectra of Nifedipine degradation product in positive ion ESI mode on an ion trap mass spectrometer (top: MS;
bottom: MS6).

in pharmaceutical analysis have been discussed by combining MS/MS experiments (and even MSn
Winger and Kemp,[35] and Ferrer and Thurman[36] experiments) with high mass resolution and accurate
have presented an in-depth discussion of TOF LC/ mass measurements, making it possible to produce
MS and the use of accurate mass measurements. Mass accurate mass-measured fragmentation maps from
measurements of sufficient accuracy (i.e., 3 ppm below trace level impurities. The potential for structure
m/z 1000) can allow the determination of elemental elucidation of trace level impurities in pharmaceutical
compositions of ions, and, when combined with MS/ products provided by such instruments is enormous,
MS experiments, can allow the construction of fragmen- and will likely serve to perpetuate the central role of
tation maps for structure elucidation of trace level ana- MS in this application.
lytes. Such accurate mass measurements are most often
acquired at high mass resolution, of which these instru-
ments are capable. Fig. 10 shows an ESI mass spectrum Other Analytical Techniques and Experiments
of II acquired on a TOF LC/MS instrument and
expanded around the molecular ion region ([M þ H]þ When GC/MS and LC/MS experiments do not
at m/z 329). The exact mass of the [M þ H]þ was allow elucidation of a trace level impurity mole-
determined to be 329.1140, confirming the molecular cular structure, it may be necessary to employ
formula of C17H16N2O5(0.8 ppm accuracy) of II. other analytical techniques. By far the most useful
At the time of this writing, new so-called ‘‘hybrid’’ of these is NMR spectroscopy. NMR and its applica-
LC/MS systems are available, which combine quadru- tions in the pharmaceutical industry have been
ples (Q) with TOF, quadruples with FTICR, and ion reviewed in detail by Lohr et al.[37] In principle,
traps with FTICR. These instruments are capable of NMR is capable of determining the connectivities
 3808 Trace Level Impurity Analysis

9.9e5 329.1140
9.5e5
[M+H]+m/z 329.1140
9.0e5
8.5e5
8.0e5
7.5e5
Intensity (counts) 7.0e5
6.5e5
6.0e5
5.5e5
5.0e5
4.5e5
4.0e5
3.5e5
3.0e5
2.5e5
2.0e5 330.1171
1.5e5
1.0e5
5.0e4

327.8 328.0 328.2 328.4 328.6 328.8 329.0 329.2 329.4 329.6 329.8 330.0 330.2 330.4 330.6 330.8 331.0 331.2 331.4
m/z (amu)

Fig. 10 High-resolution accurate mass measured positive ion ESI mass spectrum of nitrosodehydronifidipine acquired on a
TOF LC/MS system.

between all protons (H atoms) and carbons experiments for small molecule structure elucidation.
(via 13C) in an analyte molecule, thus allowing for In reality, however, NMR is very insensitive when
a complete ‘‘proof-of-structure’’ of the unknown. compared with MS. Traditionally, this lack of
Trace–Ultrasonic

Such connectivities are established by employing ‘‘2- sensitivity has required that trace level impurities be
dimensional’’ NMR experiments, which can either be isolated and enriched for NMR analysis, a time-con-
homonuclear (correlating 1H-1H connectivities) or suming and laborious process involving preparative
heteronuclear (correlating 1H-13C connectivities). HPLC or other similar techniques with extensive
Table 4 presents an overview of the most useful NMR sample cleanup. The amount of analyte required for

Table 4 An overview of NMR spectroscopy—useful NMR experiments for small molecule structure elucidation
Experiment category Experiment designationa Information content
1
1 Dimensional H No special designation Number, type, and chemical
environment of protons
13
1 Dimensional C No special designation Type, and chemical
environment of carbons
(numbers difficult to obtain)
13
Modified 1 Dimensional C APT Information regarding the
DEPT protons attached to carbon
INEPT (CH, CH2, CH3)
GATEDEC
2 Dimensional 1H (Homonuclear) COSY H–H correlation, through bond
TOCSY Long range H–H correlation,
NOESY through bond H–H correlation,
ROESY through space
Long range H–H correlation
13
2 Dimensional C (Heteronuclear) HMQC Short range H–C correlation
HMBC Long-range H–C correlation
a
Acronyms, which designate individual NMR experiments, are defined and discussed in ‘‘NMR Spectroscopy in Pharmaceutical Technology’’ in
the Encyclopedia of Pharmaceutical Technology.
Trace Level Impurity Analysis 3809

NMR structure elucidation depends on the NMR that are themselves based on MS. A systematic MS-
experiments required and the purity of the isolated based paradigm can be described as a four-step process:
material. With more than 95% pure material, several
hundreds of micrograms may be sufficient for 1H 1. Situation evaluation: Understand the impurity
NMR experiments; however, for 13C NMR experi- identification problem.
ments, several milligrams may be necessary. The reader
a. What analytical technique/method was
is cautioned that 13C NMR experiments are often
used to initially detect/report the unknown
required for a complete proof-of-structure.
impurity?
In recent years, the combination of HPLC with NMR
b. Is the impurity to be identified process
(LC/NMR) has become a reality, and the application of
related, API related, excipient related,
LC/NMR to trace level impurity identification has
container closure system related, or totally
been demonstrated.[38–40] LC/NMR experiments along
unknown (highly unusual!!!)? What are
with the development of higher sensitivity NMR probe
the physicochemical and molecular proper-
technology can potentially reduce the need for trace level
ties of related materials (e.g., the API), and
impurity isolation and sample enrichment. However, it is
how are these likely to relate to the
important to point out the following:
unknown?
c. What is the nature of the sample matrix from
1. The acquisition of 13C data sufficient to prove
which the unknown impurity was detected
the structure of a trace level impurity likely
(i.e., drug substance, drug product, etc.)?
remains beyond the routine capabilities of
currently available LC/NMR instrumentation 2. Data acquisition: Acquire appropriate LC/MS
and NMR probe technology. and/or GC/MS data depending on which
2. LC/NMR experiments require the use of deut- analytical technique is most applicable.
erated mobile phases to minimize background
signals (especially from water in 1H spectra). a. If HPLC was initially used for impurity
3. NMR spectra are a function of the solvent sys- detection and reporting, is the method
tem that the analyte is dissolved in. When an LC/MS compatible (i.e., no involatile
mobile phase constituents)? Can the

Trace–Ultrasonic
impurity profile is obtained with an HPLC
mobile phase gradient, and one wishes to com- method be redeveloped to be LC/MS
pare the NMR spectra of a trace level impurity compatible (e.g., replace sodium dodecyl
with the API in the same HPLC analysis, then sulfate with a volatile ion-pair reagent such
this complicating factor must be considered. as heptafluorobutyric acid)?
b. If the unknown impurity is API or excipient
For trace level impurity identifications, one must related, is the proton affinity of the parent
consider whether resources are better spent on amenable to positive or negative ion LC/
preparing higher quality samples for conventional MS? Which LC/MS ionization technique/
NMR experiments, or investing in and accomplishing process should be employed?
effective LC/NMR. 3. Data processing: Can a structural hypothesis be
The future holds the promise that GC/MS and LC/ developed from the available mass spectro-
MS experiments (with accurate mass measured MS/ metric information?
MS fragmentation maps) in combination with LC/
NMR (in spite of its current limitations) will be in a. LC/MS and/or GC/MS spectral interpret-
routine use for trace level impurity structure eluci- ation.
dation in the pharmaceutical industry. b. GC/MS computerized library search results.
c. Tandem MS (MS/MS) fragmentation
information.
d. Accurate mass/elemental composition
MS-BASED SYSTEMATIC APPROACH measurements.
FOR THE IDENTIFICATION OF TRACE
4. Confirmation:
LEVEL ORGANIC IMPURITIES
a. Mass spectral structural hypothesis leads to
The process for identification of a trace level organic selection or synthesis of authentic reference
impurity in a pharmaceutical product can be viewed compound.
in a systematic way. One commonly used paradigm is b. Isolation of a sufficient amount of pure
based on MS and the hyphenated analytical techniques analyte for NMR structure elucidation,
 3810 Trace Level Impurity Analysis

possibly followed by selection or synthesis b. The diode-array UV spectrum of the

of an authentic reference compound. impurity suggests that it is API related.
c. The HPLC method is LC/MS compatible
The process can be depicted as a flow chart (Fig. 11). (i.e., no involatile mobile phase constitu-
Norwood and Qui[6] and Lohr et al.[37] have presented ents).
similar flow charts. The reader should also note the d. The API has a molecular structure with
multidisciplinary case study approach presented by high proton affinity (i.e., structure contains
Alsante et al.[38] With reference to Fig. 11, consider several reduced nitrogen atoms, and also
the following two possible scenarios: one fluorine atom).
Scenario A: 2. Data acquisition:
a. Acquire a positive ion APCI LC/MS analy-
1. Situation evaluation:
sis. Determine the molecular weight of the
a. A 0.1% unknown impurity (photolysis impurity relative to the API (Note that
product) is detected and reported from an structures can sometimes be elucidated
HPLC/UV-based API chromatographic based solely on molecular weight differences
purity analytical method. from the API).

Situation: Situation Evaluation:

Impurity observed Sample Info: Sample, control, standard, method
Scenario: Process related? API related? degradation?
contamination? container/closure/packaging related?
Trace–Ultrasonic

GCable LCable LC/MS No

GCable? Develop
GC/MS Compatible?
LCable? Method

Trouble
Shooting
No Yes
No Valid Data Other Techniques: LC/MS
Obtained? Probe MS, CE/MS
NMR, IR, etc.

MS/MS
Yes No
Hypothesis or HR/MS
Needed?

Yes
Yes Yes Standard
Standard
Available? Fragments
Match?
Formula

Evaluate
Isolate Hypothesis
No Impurity No
No

Hypothesis Yes
Confirmed NMR Confirmed
Confirmed?

Fig. 11 Flow chart describing a systematic process for structure elucidation of a small molecule trace level impurity.
Trace Level Impurity Analysis 3811

b. Acquire an accurate mass measured APCI of molecular weight 121 has reacted with
LC/MS analysis. the API to form the impurity, possibly by
nucleophilic substitution.
3. Data processing:
b. The elemental composition of the impurity
a. The APCI spectrum of the impurity con- confirms the hypothesis.
firms the molecular weight to be 333 amu c. MS/MS fragmentation behavior of the
(API molecular weight 335 amu). The dif- impurities yields no definitive structural
ference in molecular weight between the hypotheses.
impurity and the API suggests double-bond
4. Confirmation:
formation via photolysis.
b. The elemental composition of the impurity a. Isolate the impurity from the drug product
confirms the molecular formula to be in sufficient quantity and purity for NMR
C19H27NO4, which compares with structure elucidation, including the acqui-
C19H26NO3F for the API. sition of 13C data.
4. Confirmation: b. NMR confirms the exact molecular struc-
ture of the impurity.
a. APCI LC/MS analyses confirm the
impurity structure to be the API with OH Obviously, one could create dozens of different sce-
replacing F (note that accurate mass mea- narios to run through this paradigm. Also, there are
surements were critical for this structure other paradigms that are in use in the pharmaceutical
elucidation, and LC/MS analyses alone industry. For example, laboratories skilled in isolation
were sufficient to solve the problem). and purification techniques might proceed directly to
b. No additional structure confirmation isolate all impurities and take them straight to NMR
required for this scenario. proof-of-structure. Other laboratories skilled in
organic synthesis might attempt to predict from chemi-
Scenario B: cal knowledge which impurities might form and syn-
thesize these to see which match observed impurities.

Trace–Ultrasonic
1. Situation evaluation: However, based on experience with current industry
practice and a review of the available scientific litera-
a. A 0.05% unknown impurity is detected and
ture, MS-based approaches similar to the one pre-
reported from an LC/UV-based drug pro-
sented here are the most widely applied.
duct chromatographic purity analytical
method.
b. The UV spectrum of the impurity suggests
that it is API related. CONCLUSIONS
c. The HPLC method is LC/MS compatible
(i.e., no involatile mobile phase constitu- The ability of modern analytical chemistry to identify
ents). and quantify impurities in pharmaceutical products is
d. The API has a molecular structure with staggering. It is now routine to elucidate organic
high proton affinity (i.e., structure contains impurity structures below the 0.1% level, which has
several reduced nitrogen atoms). led to an increased ability to detect and identify those
chemical entities of potential concern for the safety and
2. Data acquisition: efficacy of pharmaceuticals. New developments, such
a. Acquire a positive ion APCI LC/MS analy- as hybrid MS systems and LC/NMR when fully inte-
grated into the MS-based paradigm, hold the promise
sis. Determine the molecular weight of the
of increased efficiency for trace impurity identification
impurity relative to the API.
with resulting efficiency increases in the overall
b. Acquire an accurate mass measured APCI
pharmaceutical development process.
LC/MS analysis.
3. Data processing:
a. The APCI spectrum of the impurity con- ACKNOWLEDGMENTS
firms the molecular weight to be 721 amu
(Figs. 3, 4 and 8; API molecular weight The authors wish to acknowledge Boehringer Ingel-
602 amu). The difference in molecular heim Pharmaceuticals, Inc. as well as their colleagues
weight between the impurity and the API Mr. Keith McKellop and Dr. Alice Granger for their
suggests that a known formulation excipient invaluable assistance.
 3812 Trace Level Impurity Analysis

ARTICLES OF FURTHER INTEREST properties, analytical methods, and documented values.

Eur. J. Pharm. Biopharm. 1997, 43 (3), 215–242.
14. B’Hymer, C. Residual solvent testing: a review of gas-
Chromatographic Methods of Analysis: Gas chromatographic and alternative techniques. Pharm. Res.
Chromatography, p. 463. 2003, 20 (3), 337–344.
Chromatographic Methods of Analysis: High 15. Dwivedi, A.M. Residual solvent analysis in pharmaceuti-
cals. Int. J. Pharm. Excip. 2003, 33–37.
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Trace–Ultrasonic
 Transdermal Delivery: Anatomical Site Influence
Nora Y.K. Chew
Nina F. Wilkins
Acrux DDS Pty Ltd., West Melbourne, Victoria, Australia

Barrie C. Finnin
Monash University, Parkville, Victoria, Australia

INTRODUCTION of the studies were related to drugs that are currently

used in the approved TDD systems, where the purpose
Transdermal drug delivery (TDD) is known to offer of the study was to characterize the variability of a
many advantages over the oral and injectable routes particular dose form rather than to investigate the rea-
for systemic drug delivery. However, the skin is a sons for intersite variability, demonstrating the com-
complex and dynamic organ with marked barrier func- mercial desire for consistent and robust-performing
tion, which results in limitations and variations in products. However, it is important to note that inter-
the amount and nature of drugs that can be delivered subject and intrasubject variability,[2,3] experimental
across the skin and into the bloodstream. Thus a con- size,[4] and various endpoint measures (e.g., pharmaco-
tinual search for ways to optimize the permeation of kinetic vs. pharmacodynamic parameters) may have
drugs across the skin exists, in an attempt to enhance made interpretation of the results difficult. Parameters
delivery and improve patient compliance. of clinical importance will depend on the drug, but may
The recent introduction of regulatory criteria for include the amount of drug delivered to the systemic
TDD has drawn attention to the effects of anatomical circulation, the rate of drug delivery, the amount of drug
site on TDD. Understanding the factors that affect delivered to local tissues, and the incidence of adverse
Trace–Ultrasonic

site-to-site variation in TDD could enable enhanced effects such as local irritation at the site of administra-
drug delivery and offer flexibility to use multiple appli- tion. An example of the latter would be the measure of
cation sites, allowing greater patient acceptance and skin reactions[5] vs. the plasma concentration profile of
compliance and reducing drug absorption variability estradiol[6] delivered via a patch. Although the former
between and within individuals. The underlying mechan- showed no significant differences between test sites, the
isms that give rise to TDD site-to-site variation are yet latter demonstrated changes of pharmacokinetic profiles
to be elucidated. Although it is generally accepted that, at different regions of the body.
for most drugs, the stratum corneum (SC) is the rate-
limiting barrier to drug permeation, differences in drug
INFLUENTIAL FACTORS IN SITE-TO-SITE
permeation at different anatomical sites are unlikely to
TDD VARIATION
be just a function of the SC. The present review looks
at the effects of application site on TDD in light of
The rate and/or extent of drug penetration across the
the current understanding of the mechanism of drug
skin influences bioavailability. Mathematical models,
permeation. Site-to-site differences may arise because
which are designed to predict the rate and extent of
of differences in the rate and amount of drugs being
absorption, have attributed the high importance of
delivered, drug permeation into and across the skin,
SC thickness to TDD. These models, however, make
and drug clearance from the skin.
the assumption that the SC is homogenous in compo-
sition and structure, and the diffusion pathway is con-
sistent. However, as will be discussed, SC thickness is
STUDIES ON THE EFFECTS not the sole determinant of site-to-site TDD variation.
OF ANATOMICAL SITE A number of variables, including gender, age, ethnic
background, application methods, and subjects’ health
Investigations into the effects of anatomical site on TDD status may contribute to the reported regional differences
have been largely driven by regulatory compliance in TDD; however, only a limited amount of research has
assessment.[1] Table 1 lists those studies that investi- been conducted in these areas. Evidence would suggest
gated the influence of anatomical site on TDD in that drug entity (e.g., estradiol vs. testosterone; Table 1)
humans (in vivo and in vitro). Of the 38 studies, 23 and dosage form (e.g., patches[7,8] vs. spray solution[9])
showed regional TDD dependence and more than half (Text continues on page 3821.)
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-120030270
3814 Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved.
Table 1 Studies investigating the influence of anatomical site on TDD in humans (in vivo and in vitro)
Comments on site
Reference Compound Gender (age) Body sites Formulation Measures Findings dependence TDD
p
[19] Polycyclic Male (22–37) Shoulder, forehead, PAH in coal Surface disappearance Surface disappearance Discrepancy between
aromatic forearm, groin, tar ointment of PAH and excretion of PAH: shoulder > excretion results and
hydrocarbons ankle, hand of urinary 1-OH-pyrene forehead, forearm, surface disappearance
(PAHs) groin > ankle, hand could be due to low
Excretion profile number of observations
showed no significant and sensitivity of
difference between sites methodology.
Site differences might
be affected by hydration,
friction or temperature.
p
[78] 4-Cyanophenol Male Forearm (ventral), Patch (8
2 cm) Tape stripped, up to Partition coefficient:a Contributing factors
(CP) (Chinese; back, thigh, with sponge (Softwick 20 times in 2–3 min, Back > abdomen > could include thickness
Transdermal Delivery: Anatomical Site Influence

22–25) leg, abdomen IV sponge; J&J TX) of SC at the thigh > forearm > and number of cell
containing drug application area. leg layers, and lipid
saturated in content of SC.
aqueous solution
Drug concentration Diffusion coefficient:a Regional variation in
on each tapestrip Leg > forearm > SC transport of CP
was determined by back > thigh > might be influenced
ATR-FTIR. abdomen by the intrinsic diffusivity
of SC; while both
thermodynamic and
kinetic differences among
different anatomical
skin sites might attribute
to the differences
observed for CM.
Permeability The observed differences
coefficient:a might not be equally
Back > forearm > reflected in systemic
thigh > leg > absorption.
abdomen
Cimetidine Partition coefficient:a
(CM) Back > forearm >
abdomen > leg > thigh
Diffusion coefficient:a
Back > thigh >
forearm > leg > abdomen
Permeability coefficient:a
Back > forearm >
thigh > leg ¼ abdomen
(Continued)
3815

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 Trace–Ultrasonic

Table 1 Studies investigating the influence of anatomical site on TDD in humans (in vivo and in vitro) (Continued)
3816

Comments on site
Reference Compound Gender (age) Body sites Formulation Measures Findings dependence TDD
p
[79] Ethyl nicotinate, Male Forehead, presternal Compound dissolved Responses of erythema In general, forehead, Difference could be
privine base, (20–60) area, interscapular in 95% alcohol and whealing presternal area, and due to blood flow, follicular
histamine base region, flexor, extensor back demonstrated numbers at particular site,
surfaces of forearm, greater absorption than and nature of penetrant.
pretibial, peroneal the extensor surface of
surface of leg the forearm or either
of the areas tested on
the leg. Absorption
on the forearm was
also higher than the leg
p
[21] Parathion Male Forearm, palm, Compound dissolved in Excretion profile Total excretion of Hair follicular areas
foot ball, abdomen, acetone was applied to parathion: forearm < appeared to have higher
hand dorsum, fossa various sites palm < foot ball < drug penetration but
cubitalis, scalp, jaw abdomen < hand dorsum < authors suspected that
angle, postauricular, fossa cubitalis < scalp < differences in the skin
forehead, ear canal, jaw angle < postauricular < other than the hair itself
axilla, scrotum forehead < ear canal < might be important.
axilla < scrotum Lack of barrier property
and ‘‘absorbent sponge’’
behavior of the scrotum
might account for
the findings.
p
[27] Acetylsalicylic Male Arm, abdomen, Compound in Regional distribution All compounds appeared to More sebaceous
acid, benzoic (28  2) postauricular, ethylene glycol (measured by tape follow the ranking: arm  gland at forehead, but
acid, caffeine, forehead water/TritonÕ
100 stripping) and abdomen < postauricular < possibly an overestimation;
sodium benzoate excretion of drug forehead. closeness of capillaries to
Compounds appeared to be skin surface could be a
2
more permeable at determinant factor.
forehead than at arm or
abdomen (except caffeine).
p
[45] Acetylcholine Male and 12 different sites Iontophoresis delivering Flux changes upon Baseline flux and Thinner SC at forearm
(AC) Female at forearm AC solution application of AC vasodilatation response than palmer
(21–42) were site dependent.
Palm sites showed a higher
baseline flux, but no
vasodilatation in response
to iontophoresis.
Volar forearm, dorsal hand
and finger sites showed lower
site-dependent baseline flux,
but did vasodilate.
p
[80] Lidocaine (L) Male (20–30) Scrotum, Cotton pledgets saturated Time required to produce Anaesthetic effect: 3.6 min Skin barrier of scrotum
abdomen with L in water/IPA/ anaesthetic effect for scrotum vs. 1 hr þ for is different from other
glycerine abdomen body sites
Transdermal Delivery: Anatomical Site Influence
 Salicylic Male skin Scrotum, SA in dilute Penetration of SA Penetration time: scrotum
acid (SA) (55–72) abdomen ferric chloride 15 min vs. abdomen 2 hr þ
[8] Lidocaine (L) Male and Back, chest, IDDS iontophoresis L concentration in plasma Plasma L in all subjects
and epinephrine female (5–15) dorsum of hand, and skin reaction was <10 ng/mL irrespective
antecubital fossa of sites.
(randomly chosen) No erythema or edema from
L was found.
Erythema associated with the
anode appeared to be more
prominent at the chest or back.
p
[81] Norelgestromin Female (20–45) Abdomen, EvraTM patch Pharmacokinetic AUC: Abdomen
(NGMN) buttock, arm, (RW Johnson parameters < arm ¼ buttock ¼ torso.
torso Pharmaceutical
Research Institute, NJ)
Ethinyloestradiol Ortho Abdomen application
Transdermal Delivery: Anatomical Site Influence

(EE) EvraTM patch is not bioequivalent,

but therapeutically
equivalent to that
applied at the
other sites.
p
[7] Clonidine (CL) Male and Arm, thigh, chest Catapres-TTS patch CL concentration in CL concentration in
female (20–39) (Boehringer Ingelheim, blood and urine plasma and urine samples:
Germany) chest > upper arm > thigh.
Correlation found between
CL release from patch
and AUC.
[82] Clonidine (CL) Male (18–45) Chest, arm Catapres-TTS patch CL concentration Similar pharmacokinetic
in plasma profile between sites
p
[83] Clonidine (CL) Male (26–41) Right chest, Patch Pharmacokinetic Plasma concentration
left arm, upper parameters of CL, Cmax, AUC: Left
abdomen arm were greater than
right chest and abdomen.
Blood pressure lowering:
arm > abdomen.
[5] Estradiol (E2) Female Various sites of ClimadermÕ patch Skin reactions No difference in the
(Brazilian; abdomen: upper, (Wyeth-Ayerst, PA) number of incidence
35–60) middle, and lower and severity of reactions
between regions
[13] Estradiol (E2) Female Abdomen, buttocks, Esteraderm TTSÕ E2 conjugates in urine No significant difference Berner and John[10]
(47–59) upper thigh, lower (Ciba-Geigy LTD, between sites. Intersubject proposed that insignificant
back, lateral thorax, Switzerland) patch variability was high. difference might be due to
upper arm the contribution of ethanol
and a rate-controlling
membrane to control
estradiol delivery.
(Continued)
3817

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 Trace–Ultrasonic

Table 1 Studies investigating the influence of anatomical site on TDD in humans (in vivo and in vitro) (Continued)
3818

Comments on site
Reference Compound Gender (age) Body sites Formulation Measures Findings dependence TDD

If data were subjected to

paired t-test analysis, upper
thigh values were significantly
different from those at abdomen.
Bioequivalence was established
for all sites except the upper thigh.
p
[6] 17-b Estradiol Female (45–70) Abdomen or ClimaraÕ patch Pharmacokinetic AUC: Buttock > abdomen. Variation might be due
(E2) buttocks parameters Cavg: buttock > abdomen. to SC thickness, lipid
content, regional blood
Tmax: buttock < abdomen.
flow, age, and race.
Greater peak and trough
for buttock.
p
[3] 17-b Estradiol Female Abdomen or buttocks FemPatchÕ patch Pharmacokinetic Mean serum E2 Skin thickness, blood
(E2) (Cygnus Therapeutic parameters concentration: flow, lipid content,
Systems, CA) buttocks > abdomen and number of
hair follicles
p
[84] Estradiol (E2) Not specified Buttocks, abdomen ClimaraÕ (Berlex Pharmacokinetic AUC, Cmax, Tmax: Lipid content, dermal
Laboratory, Inc.) parameters buttocks > abdomen. thickness, hairiness, and
Bioavailability was superficial blood circulation
more variable at
abdomen.
[85] Estradiol (E2) Not specified Arm, thigh, OesclimÕ 50 Serum levels of E2 No difference
abdomen, buttocks (EsclimÕ/EsclimaÕ; between sites
Groupe Fournier)
p
[86] Estradiol (E2) Male cadaver, Hip, abdomen, Not specified Permeation rate of E2 Permeation rate:
full thickness chest, thigh, hip < abdomen,
and dermis arm, back chest, thigh, arm, back
[87] Fentanyl (F) Female, full Breast, abdomen F in aqueous solution Transdermal flux No difference observed
thickness or in patch, was applied between test sites
on skin mounted to
diffusion cell
[88] Fentanyl (F), Human Foot, chest, Saturated solution of F Permeability coefficient No difference observed
sufentanil (SF) cadaver skin thigh, abdomen and SF applied on the between test sites
skin mounted to
diffusion cell
p
[2] Hydrocortisone Male Forearm (ventral), HC in acetone Excretion profile of HC Percent dose found in Areas with large or
(HC) forearm (dorsal), urine: Forearm (ventral) < numerous hair follicles or
foot arch (plantar), forearm (dorsal) < foot thin stratum corneum
angle (lateral), palm, arch (plantar) < ankle might result in higher
back, scalp, axilla, (lateral) < palm < drug absorption.
forehead, jaw back < scalp < axilla < Schenkel, Barlier, and
angle, scrotum forehead < jaw angle < Riera,[11] believed that
scrotum. non-occlusive application
Transdermal Delivery: Anatomical Site Influence

HC was undetectable at heel. might result in lost of

steroid from skin surface.
 p
[89] Ketoprofen (K) Male Back, arm, and knee Gel (solution form Pharmacokinetic Relative bioavailability Alternation in the
was used as control) parameters was similar between sites. thickness and diffusivity
The plasma ketoprofen of the SC in the
Cmax for gel applied to the various regions
back and arm was similar
(p > 0.05), but Cmax was
lower at the knee (p < 0.05).
Cmax range was similar for gel
treatments on the back and
arm, but longer for the
knee treatment.
p
[90] Levonorgestrel Female Abdomen, arm, LNG/E2 TCDS patch Pharmacokinetic Cmax, Tmax, and AUC were The reservoir effect of
(LNG) haunch parameters different between sites. After LNG might lead to
removal of patch, LNG/E2 similar pharmacokinetic
AUC, and T1/2 of abdomen profiles between sites.
were greater than that
of the arm and haunch.
p
Transdermal Delivery: Anatomical Site Influence

[39] Methyl Male and Forehead, forearm, MN in aqueous solution Laser Doppler flowmetry AUC and Rmax (response AUC and Rmax
nicotinate female (18–40) palm delivered via a patch response time): forehead > differences might be
(MN) forearm > palm due to follicular density
Tmax and T50% (time for differences between sites.
maximum response to decrease Tmax and T50% differences
to Tmax/2): forearm > could be due to cell size,
forehead > palm. lipid content at
various sites.
[91] Nicotine (NIC) Male (21–46) Upper back, upper Nicoderm (Alza Pharmacokinetic No difference between
outer arm, upper chest Corp., CA) profile the test sites
[74] Nitroglycerin Male and Arm, chest, thigh Ointment Hemodynamic and Equivalent hemodynamic Diffusivity vs. SC thickness;
(GTN) female (58–77) (Kremers-Urban GTN concentration effects and AUC between sites. hydration and reservoir
Co., Milwaukee) over time effects may affect
Site was covered by bioavailability
occlusive dressing
p
[76] Nitroglycerin Human (25–33) Forehead, chest, NitrolÕ patch Heart rate and Systolic blood pressure
(GTN) ankle as (chest or (Kremers-Urban Co.) blood pressure reduction and onset of
ankle control). Applied site was covered changes: forehead ¼
with tape chest > ankle
[92] Paraquat (PQ) Human Leg, forearm, hands PQ in aqueous solution Urine PQ concentration No difference between sites
vs. time
p
[93] Scopolamine Human SC Back, chest, Not cited Penetration flux Permeation flux: Less dense and thinner
(SPA) forearm, thigh postauricular > back > stratum corneum; greater
chest ¼ stomach > number of appendages;
forearm > thigh deep indentation of
dermal papillae into
epidermis could bring
capillaries closer to the
skin surface, thus increasing
surface temperature and
permeation.
3819

(Continued)

Trace–Ultrasonic
 Trace–Ultrasonic

Table 1 Studies investigating the influence of anatomical site on TDD in humans (in vivo and in vitro) (Continued)
3820

Comments on site
Reference Compound Gender (age) Body sites Formulation Measures Findings dependence TDD
p
[94] Methyl salicylate Human Abdomen, forearm, MetsalTM (Raker, Plasma concentration Abdomen > forearm > SC thickness, hair follicles
instep, heel, plantar Sydney, Australia) instep > heel > plantar
[15] Salicylic acid Human SC Abdomen, leg Propylene glycol Penetration flux Differences in flux between Correlation found between
(SA) containing drug sites were observed penetration flux and lipid
solution applied to content; no correlation with
skin mounted on skin thickness and cell layers.
in vitro diffusion cell
[95] Testosterone (T) Male (25–69) Upper buttocks, D-Trans (Alza Corp.) Pharmacokinetic Similar pharmacokinetic
upper arm, back parameters profiles between sites
[4] Testosterone (T) Male (26–59) Regimen 1—single Androgen gel (Besins Pharmacokinetic Similar T serum level and AUC Limited number of
site: left arm/shoulders Iscovesco, France) parameters between the two regimens. subjects in the study
Regimen 2—separation (hydroalcoholic gel) could give rise to the
sites: left and right similar T concentration
arms/shoulders and and AUC between the
left and right abdomen two regimens
p
[9] Testosterone (T) Female Abdomen, forearm MDTSTM spray solution Pharmacokinetic Average and maximum serum
parameters concentrations of free and
total T were significantly
higher after application to the
forearm compared to
the abdomen
p
[77] Testosterone (T) Male (21–65) Abdomen, back, chest, Androderm TTD Pharmacokinetic Bioavailable T: back > Skin permeability,
shin, thigh, upper arm (TheraTech, UT) parameters thigh > upper arms > cutaneous blood flow,
(TTD contained water, abdomen > chest > skin and/or the degree of
ethyl alcohol, glycerin, adhesion between
glycerol monooleate, system and skin
methyl laurate,
gelling agents
[96] Oxybutynin Male and Abdomen, OXY TDS (Watson Pharmacokinetic Bioequivalent OXY
(OXY) female (20–77) buttocks, hip Laboratories, Inc., parameters absorption between sites
UT) patch
[97] Tri-n-butyl- Male cadaver, Scrotum, postauricular, Aqueous solution TNBP concentration in No difference between Small sample size
phosphate full thickness thigh, scalp, instep, containing TNBP was receptor solution regional areas could affect results
(TNBP) skin anterior forearm, applied to skin mounted
plantar on in vitro diffusion cell
a
Notation as per Ref.[78].
p
( ) Study where site dependence TDD was observed.
AUC ¼ area under the concentration–time curve (unless otherwise stated); Cmax ¼ maximum concentration; Cavg ¼ average concentration; Tmax ¼ time to reach maximum concentration;
T1/2 ¼ half-life.
Transdermal Delivery: Anatomical Site Influence
Transdermal Delivery: Anatomical Site Influence 3821

do not participate in site-to-site differences of transder- 5

Dermis
mal bioavailability (TDB). That said, Berner and John[10]
4 Epidermis
had previously pointed out that insignificant differences

Thickness (mm)
between application sites for the delivery of estradiol[11] 3
could be due to the contribution of ethanol and a rate-
2
controlling membrane that potentially influenced the
delivery of the drug. Thus the effect of dosage form on 1
site-to-site variation may require further investigation.
The major factors that appear to influence TDD site 0
variation include SC structure, (epi)dermal structure, Scalp Forehead Back Abdomen Thigh Wrist Palm
lipid content and structure, cutaneous blood flow,
Fig. 1 Varying skin thicknesses at different body regions.
and skin occlusion.
(From Ref.[18].)

SKIN STRUCTURE To emphasize this, Behl et al.[23] reported that only in

the absence of the SC were differences in the penetration
Surface Topology of hydrocortisone observed between the abdomen and
the dorsal sites of rats. This was believed to be due to
Although differences in surface characteristics, such the thicker dorsal dermis as compared with that of the
as smoothness and the presence of hair, might be abdomen.
expected to influence TDB from transdermal dose
forms, there are limited supportive data. Evidence sug-
gests that differences in sebum sheet thickness between Cellular Layers and Cell Sizes
different sites may contribute to differences in the
delivery of drugs to the SC.[12] It is also reasonable Regional variation in the number of SC cellular layers
to assume that soap and other topically applied pre- and cell sizes has also been documented.[15,16,24,25] The
parations such as moisturizers might similarly affect former was thought to account for the variation in
SC thickness.[16] To date, the contribution of SC cellu-

Trace–Ultrasonic
TDB, as they are unlikely to be uniformly applied to
all areas of the body. That said, Watkinson et al.[13] lar layers and cell size to TDD variation remains
concluded that the axilla had a physiologically reduced unclear.[15,26] It has been proposed that, depending
barrier function because an exaggerated application of on the SC thickness, the variation in cell size could
antiperspirant had no effect. lead to variation in the volume of intercellular space,
which may act as the molecular ‘‘reservoir,’’[27] thus
influencing drug penetration and sorption. Because it
Thickness
is thought that most drugs permeate the SC by diffus-
ing through the intercellular lipid matrix, the length of
The laws describing diffusion through a membrane
this pathway is more likely to reflect relative per-
accord a major role to membrane thickness. In TDD,
meation rates than the thickness of the SC. The size
the membrane of interest is the SC, which is generally
of the corneocytes and their structural arrangement
considered to be rate-limiting in transdermal permeation.
within the SC may have a marked effect on the effec-
Previous observations in the variations of SC thickness
tive length of the diffusion pathway, thus influencing
at different body sites have been documented with
transport of drug across the skin and absorption into
supportive measurements.[14–18] Even though such varia-
the blood circulation.
tions in the SC have been used to explain the site depen-
dence of TDD (Table 1), no definite association has been
established in humans[16–19] or animals.[20] For instance, Lipid Content
although the SC thicknesses of the arm and scrotum were
found to be comparable,[18] the amount of parathion[21] The effect of delipidization in compromising the SC
or hydrocortisone[2] absorbed was significantly different barrier function[28–30] and alteration of solute uptake[26]
between the two sites. Thus the dermis thickness should clearly implicates the involvement of the lipid pathway
also be considered as it shows regional differences 1000 in the transdermal delivery of drugs. Changes in drug
times greater (millimeter range) than the measurable permeation through the skin in diseases that alter the
but minor regional variations (changes in micrometer lipid content of the skin have also substantiated the
size range) of the SC (Fig. 1). However, the dermis also importance of lipid to drug penetration. For instance,
has the capability to retain drugs.[22] In TDD, the drug ichthyosis is a genetic skin disease that features hyper-
molecule is required to diffuse through to the papillary keratosis, which results in the thickening and scaling
dermis or even deeper for systemic absorption. of the SC due to the accumulation of cholesterol
 3822 Transdermal Delivery: Anatomical Site Influence

sulphate, an amphipathic lipid in the epidermis.[31] This resulted in an increase in drug concentration of both
alteration in the lipid matrix is accompanied by a more lidocaine and salicylic acid in the underlying tissues.
rapid drug permeation. Similarly, Morgan, Renwick, and Friedmann[43] had
The lipid composition of the SC varies between the reported a 15-fold difference in the recovery of
different regions of the body.[32–35] A study by Elias penciclovir through the human skin with and without
et al.[17] demonstrated a quantitative relationship cutaneous blood flow.
between the rate of percutaneous transport of salicylic Blood flow within the skin has been shown to vary
acid across the SC at two different body sites (abdo- at the different body sites.[39,44,45] A tendency for skin
men and leg) and the lipid weight percent at the blood flow to decrease gradually from the upper part
corresponding sites. The results showed that the lower of the body to the lower part of the body was observed,
the lipid content, the higher the penetration rate of in the order of face, anterior chest, deltoid, hands, and
salicylic acid, which appeared to be in line with the role feet. Studies investigating the effect of regional blood
of the lipid pathway to the non-polar solute pene- flow variation on TDD are limited and inconclusive,
tration.[26] A more in-depth study by Lampe et al.[34] which appears to be due to the fact that the drug can-
also found the same variations in lipid weight percent, didates investigated also possessed vasoactive activity;
with face > abdomen > leg > plantar SC in the thus the actual contribution of blood flow toward
exact inverse of their known permeability.[36] regional differences in TDD may have been masked.
For example, Tur, Maibach, and Guy[39] showed that
the response produced by the topically applied methyl
Appendageal Pathway nicotinate did not follow the corrected regional base-
line blood flow of the skin trend (forehead >
Recently, Roberts, Cross, and Pellett[37] reviewed the palm > forearm) because the Tmax response and
contribution of appendageal pathway to TDD. Among area-under-the-curve (AUC) values were the highest
the four appendages of the skin (sweat glands, hair at the forehead and the lowest at the palm, with the
follicles, and the associated sebaceous glands and nails), forearm results falling somewhere between the other
hair follicles and the associated sebaceous glands are data sets. Another report by Gardner-Medwin et al.[45]
the only appendageal pathways that are believed to showed an unusual relationship between the blood
Trace–Ultrasonic

contribute to drug permeation. The importance of this flow of the different regions of the skin and the corre-
route is usually dismissed because the appendages sponding vasodilatation response after the administra-
occupy only 0.1–1.0% of total skin area. Studies on tion of acetylcholine. It was found that the area
TDD via the appendageal route are scarce; however, (palmer) that had the higher microcirculation flow,
a few studies have demonstrated a potential contri- in the absence of drug, showed no vasodilatation in
bution of this route to the overall flux of drugs.[38,39] response to iontophoresis of acetylcholine; whereas
Because the distribution of appendages is not uniform the areas (volar forearm, dorsal hand, and finger) that
across the body, it is likely for these drugs to experience had the lower site-dependent microcirculation flow
site variation. demonstrated vasodilatation from acetylcholine. Thus
the lack of response to iontophoresis at the palmer site
was interesting in spite of the higher microcirculation
CUTANEOUS BLOOD FLOW flow at the site. In an attempt to explain this, it was
considered that during the process of iontophoresis,
Systemic absorption and tissue distribution of trans- the drug molecule is still required to overcome the SC
dermally applied drugs are believed to be partly barrier. Thus the thicker SC of the palm, as compared
influenced by cutaneous perfusion below the site of with the forearm, dorsal hand, or finger, provided a
application, and the orientation and accessibility of greater resistance for acetylcholine to pass through
local blood vessels and tissues.[40] The other contribu- prior to reaching the blood circulation.
ting factor is considered to be drug plasma-binding Secondary factors such as humidity, temperature,
characteristics, which are dependent on the chemical skin maturation, and diurnal rhythms, which alter SC
nature of the drug, and will not be further discussed composition, physical structure, and cutaneous blood
in this review. Studies involving animals,[41,42] and more flow, need to be considered when examining the causes
recently studies with humans,[43] have demonstrated of site-to-site variation in TDB.
that cutaneous blood flow affects drug pharmaco-
kinetics and the relative processes of local and systemic
solute distribution, as well as drug clearance following Humidity and Temperature
application. It was shown that the coadministration
of a vasodilator (phenylephrine), which was believed to Low air humidity (60%)[46] or high temperatures
limit the removal of solutes by the dermal circulation, are thought to dry out the SC[47,48] and subsequently
Transdermal Delivery: Anatomical Site Influence 3823

alter barrier function.[47–50] Such changes could be dif- Skin Maturation

ferent at the various body sites.[48] Hydration is known
to thicken the SC as a result of cells swelling, and cause The human skin changes dramatically with age. Among
alterations in the arrangement of lipids, which may the vast number of skin characteristics that have been
reduce the diffusional resistance of the SC. At the shown to change with age, the reduction in skin
ultrastructural level, high humidity has been shown hydration level,[60–63] change in the lipid content,[32,56,64]
to disrupt the SC and the lipid lamellae.[51,52] and decrease in dermal circulation[65–68] are likely to
The effect of seasonal influences on physiological have an impact on TDB. The extent of these changes
processes occurring in the skin appears to be extremely was also found to be different at different body sites.
variable. Previous investigation showed that damage to However, there are no published data on the regional
the skin barrier function by an irritant was more differences in transdermal delivery profiles with age
significant in winter than in summer.[53] Furthermore, for humans. A recent report by Cua, Willhelm, and
a more recent study[54] showed that seasonal-induced Maibach.[69] showed a variation in response to sodium
changes in the SC may also be related to the anatom- lauryl sulfate (SLS) irritation with age, with the elderly
ical site, with the most significant changes observed displaying a different and weaker regional response than
in summer when humidity and temperature are at the the young. Although a number of anatomical regions
highest. Of the three test sites, the forearm and calf were investigated, the potential decline of immune
areas showed comparable characteristics of hydration, function in the elderly group must also be considered.
transepidermal water loss (TEWL), and desquamation.
These were found to be higher than those from the face
region. During summer, the levels of these parameters Circadian Cycles
were elevated at all sites except those at the face.
Seasonal changes in SC lipids (such as fatty acid, cer- Significant changes in skin composition at different
amide, and cholesterol) have also been observed,[55,56] times of the day have been observed in a number of
and the degree of changes was found to be different studies. Changes with time to variables such as TEWL,
at different body sites. Thus it would appear that skin surface pH, skin temperature, and skin color have
seasonal changes in the SC may contribute to the been detected at most anatomical sites.[70,71] However,

Trace–Ultrasonic
anatomical site variation of TDD. SC hydration levels were found to be unaffected.
Although exercise is known to change the body tem- Yosipovitch and Farrar recently reported a statistically
perature and blood flow of the skin, this does not significant circadian rhythmicity in TEWL, pH, and
always translate to changes in TDB. Barkve et al.[57] temperature on the forearm, forehead, and shin, but
and Vanakoski et al.[58] reported an increase in the not on the upper back. The study showed that the
plasma concentration of glyceryl trinitrate (GTN) after water evaporation rate increased significantly in the
exercise or sauna in patients wearing transdermal evening at all sites, but was minimal during the day.
patches. Vasodilation of the cutaneous skin blood However, this did not appear to be related to the diur-
flow as a result of exercise or sauna was the proposed nal variation in sweating. The difference in TEWL on
explanation to this finding. The minimal correlation the forearm and forehead was thought to be due to the
observed between the dermal temperature and GTN dissimilar circadian rhythms at different regions of the
uptake was believed to be related to the dilation of body.[72] Oscillation of body temperature, which has
blood vessels and sweating during exercise. In contrast, been known for centuries, was thought to be related
the delivery of norelgestromin (NGMN) and ethinyl to the variation of heat loss during the day.[73] How-
estradiol (EE2) via a patch was reported to be unaf- ever, individuals or populations in similar time zones
fected by exercise,[59] with the concentration of NGMN do not necessarily have similar circadian rhythm tem-
remaining unchanged during the study period. EE2 perature patterns.
demonstrated a slight increase in serum concentration
during the first 48 hr of the patch application, which
then decreased during the subsequent 5 days of the SKIN OCCLUSION
wearing period. Because there was no observable
change for NGMN, the slight change of EE2 during It has long been recognized that physical occlusion of
the first 48 hr of the wearing period could not be the skin could increase the water content of SC by
explained solely by the change in blood flow. The up to 50% and the surface temperature from 32 C to
potential effect of drug depletion from the patch was 37 C.[37] These changes were considered to change the
not mentioned. In addition, it is possible that any effect thermodynamics of drug penetration,[74] thus enhancing
of exercise on the permeability of the hormones may the delivery of certain drug candidates.[75]
have been masked by the rate-controlling release from Regional variation of TDD had been observed even
the patch. in the presence of physical occlusion of the skin, as
 3824 Transdermal Delivery: Anatomical Site Influence

demonstrated in previous studies.[3,7,76,77] A more differences in the time of maximal excretion paralleled
recent study in pigs by Qiao, Chang, and Riviere[20] these results. It was proposed in this study that the
reported that the trend in regional differences of drug vascular anatomy was important because the non-
absorption between the occluded and non-occluded occluded data showed that the ventral abdomen had
systems was different. Formulations containing the lowest absorption and was the only skin site supplied
2,6-[ring-14C] parathion were applied at four different by direct cutaneous vessels rather than musculocuta-
swine skin sites. Urinary and fecal drug excretion neous arteries.
(% dose) at 168 hr and the time of maximal drug
excretion were examined. Physical occlusion gave rise
to an increase in the dose delivered at all sites (Fig. 2). CONCLUSIONS
Under occluded conditions, the TDB was lower in the
shoulders than in the other regions. In contrast, under Interpretation of the published data to elucidate the
non-occluded dosing, there was more variation reasons for site-to-site variation in TDB is complicated
between the sites, with the TDB from the abdomen by the interaction of multiple factors. If the factors
being significantly lower than the other sites. The that affect anatomical variation in TDD can be under-
stood, then it may be possible to improve TDD by
decreasing site-to-site variability or by improving bio-
availability by deliberately manipulating the factors
controlling delivery. The present review demonstrates
the importance of skin thickness, lipid content,
cutaneous blood flow, skin occlusion, and change in
humidity and temperature (in effect of, e.g., exercise
and seasonal changes). Currently, there is insufficient
evidence to demonstrate the contribution of cellular
layers, arrangement of cells, cell sizes, skin appendages,
skin maturation, and circadian cycles to the anatom-
ical variation of TDD. Future studies may encompass
Trace–Ultrasonic

the contribution of dermal thickness and composition

at different regions of the body; the extent of drug
retention in TDD; the organization of corneocytes
at the different body sites, which in turn relates to
the transport of drugs; the influence of blood flow at
various body sites, particularly studies utilizing drug
candidates that have little or no interference of the
actual blood flow; and the dosing frequency (e.g.,
single vs. multiple, finite vs. infinite dosing) of the
site-to-site differences of TDD.

ACKNOWLEDGMENT

The authors would like to thank Dr. Tim Morgan for

his guidance in this review.

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human stratum corneum ultrastructure. J. Invest. Dermatol. availability of nitroglycerin ointment in congestive heart
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Trace–Ultrasonic
 Transdermal Delivery: Sonophoresis
Samir S. Mitragotri
Department of Chemical Engineering, University of California,
Santa Barbara, Santa Barbara, California, U.S.A.

Hua Tang
TransForm Pharmaceuticals, Inc., Lexington, Massachusetts, U.S.A.

E. Daniel Blankschtein
Robert Langer
Department of Chemical Engineering, Massachusetts Institute of Technology,
Cambridge, Massachusetts, U.S.A.

TRANSDERMAL DRUG DELIVERY Amplitude of ultrasound waves can be represented in

terms of peak wave pressure (in Pascals) or in terms of
Systemic as well as topical delivery of drugs via the intensity (in the units of W/cm2). Ultrasound can be
transdermal route is limited by the low skin permeability applied either continuously or in a pulsed manner. In
which is attributed to the stratum corneum (SC), the the latter case, an additional parameter, duty cycle, is
outermost layer of the skin.[1] The SC consists of disk- required to characterize ultrasound application. Duty
like dead cells (keratinocytes) containing keratin fibers cycle is the fraction of time for which ultrasound is ON.
and water, surrounded by densely-packed lipid bilayers. Ultrasound is generated using a device referred to as
The highly-ordered structure of the lipid bilayers confers a sonicator. It consists of an electrical signal generator
a highly impermeable character to the SC. A variety of which generates an electrical AC signal at the desired
approaches have been suggested to enhance transdermal frequency and amplitude. This signal is applied across
Trace–Ultrasonic

drug transport. These include: 1) use of chemicals to a piezo-electric crystal (transducer) to generate ultra-
either modify the skin structure or to increase the drug sound. The thickness of the piezo-electric crystal is
concentration in the transdermal patch;[2,3] 2) applica- selected so that it resonates at the operating frequency.
tions of electric fields to create transient transport path- Sonicators operating at various frequencies in the
ways [electroporation][4,5] or to increase the mobility of range of 20 kHz to 3 MHz are available commercially
charged drugs through the skin [iontophoresis];[6] and and can be used for sonophoresis.
3) application of ultrasound [sonophoresis].[7–56] If a sonicator operating at the desired frequency is not
Sonophoresis was shown to enhance transdermal drug available commercially, it is possible to assemble one
transport about half a century ago by Fellinger and using commercially available signal generators, ampli-
Schmidt[16] who showed that application of ultrasound fiers, and transducers. Such sonicators operating at fre-
increases transport of hydrocortisone across the skin. quencies of 10 MHz and 16 MHz have been assembled
Following this study, attempts were made to enhance by Bommannan et al.[11] (For a discussion of the relevant
transdermal transport of more than 15 drugs including methods for making a custom sonicator, see Ref.[11].)
steroidal anti-inflammatory drugs such as hydrocorti- For sonophoretic delivery, the desired drug is dis-
sone, dexamethasone; non-steroidal anti-inflammatory solved in a solvent and applied on the skin. Ultrasound
drugs such as salicylates and ibuprofen; anesthetic agents is applied by contacting the transducer with the skin
such as lidocaine; and proteins such as insulin. This chap- (Figs. 1A–C) through a coupling medium to ensure a
ter provides a review of these studies with emphasis on proper contact between the transducer and the skin. This
associated techniques, mechanistic studies, and safety. medium can be the same as the solvent used to dissolve
the drug or it can be a commercially available ultrasound
coupling gel (for example, Aquasonic, Polar, NJ).
GENERATION AND APPLICATION OF
ULTRASOUND FOR SONOPHORESIS Transmission of Ultrasound from the
Transducer to the Skin
Generation of Ultrasound
Transmission through the medium
Ultrasound is a sound wave possessing frequencies
above 20 kHz.[57,58] These waves are characterized by Ultrasound requires a coupling medium for trans-
two main parameters: frequency and amplitude. mission from the transducer to the desired tissue.
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-100200017
3828 Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved.
Transdermal Delivery: Sonophoresis 3829

Ultrasound
Transducer
Dissolve
Compressure Drug Solution
Skin

Buffer
Reservoir
Compressure Ultrasound
Transducer
Starter

Drug Reservoir

1A: In Vitro Experimental Set-up

Ultrasound
Transducer

1B: In Vivo Experimental Set-up

Drug in Coupling
Medium

Trace–Ultrasonic
Patient's Hand

1C: Clinical Experimental Set-up

Fig. 1 Experimental set-up for sonophoresis delivery.

The coupling medium should result in proper trans- indicated by the absorption coefficient (a). The extent
mission of ultrasound from the transducer to the skin. of absorption is given by the following equation.
The transmittive properties of a medium are indicated
by its acoustic impedance (Z). A coupling medium is
fðtÞ ¼ 1  expðatÞ
appropriate for sonophoresis if its acoustic impedance
(Z) is comparable to that of skin (1.6
106 kg/m2/s).
where f(t) is the fraction of ultrasound intensity
Z-values for various materials can be found in
absorbed as the ultrasound beam propagates in a
Refs.[57,58]. For example, water has a Z-value of
medium with absorption coefficient a and thickness t.
1
106 kg/m2/s and is a reasonable coupling agent.
An estimation of a-values for various materials at vari-
Z-values for several media are listed in Table 1.
ous ultrasound frequencies may be found in Refs.[57,58].
In the case of water, the a-value is 0.0006 at an ultra-
sound frequency of 1 MHz, suggesting that a 1 cm
Absorption of ultrasound thick column of water absorbs less than 0.1% of ultra-
sound (1 MHz) intensity, i.e., water is a reasonable
Every medium absorbs ultrasound to a certain extent. coupling medium. Absorption coefficients for several
The ability of a medium to absorb ultrasound is media are listed in Table 1.
 3830 Transdermal Delivery: Sonophoresis

Table 1 Acoustic impedances and absorption coefficients Therapeutic frequency ultrasound (1–3 MHz): This
of materials is the most commonly used ultrasound frequency range
Absorption for sonophoresis. Specifically, over 90% of the previous
Acoustic coefficient studies of sonophoresis have been conducted using
impedance, (a) at 1 MHz therapeutic ultrasound. A summary of these studies is
Material Z (kg/m2/s) (cm1) provided in Table 2. Interestingly, in the therapeutic
Water 1.5
106 0.0006 frequency range of 1–3 MHz, frequencies closer to
Blood 1.6
10 6
0.028 1 MHz have been preferably used for sonophoresis.
No reason has been given by investigators for the use
Bone 6.3
106 3.22
6
of this particular frequency. Mitragotri et al.[36]
Skin 1.6
10 0.62 reported that the sonophoretic enhancement in the
Fatty tissue 1.54
106 0.14 therapeutic frequency range varies inversely with ultra-
6
Muscle 1.6
10 0.76 sound frequency. They found that while 1 MHz
Air 0.0004
106 2.76 ultrasound enhances transdermal transport of estradiol
(From Hoogland, R. Ultrasound Therapy; Ernaf Nonius: Delft, across human cadaver skin in vitro by 13-fold, 3 MHz
Holland, 1986.) ultrasound at the same intensity induces an enhance-
ment of only 1.5-fold. They further hypothesized that
the observed inverse dependence of sonophoretic
enhancement on ultrasound frequency occurs since
Ultrasound reflection cavitational effects, which are primarily responsible
for sonophoresis, vary inversely with ultrasound
Ultrasound is reflected at the boundary of two media frequency.[37,59]
possessing different acoustic impedances. 99.99% of High-frequency ultrasound (above 3 MHz):
ultrasound is reflected at the air-water boundary when Bommanan et al.[11,12] performed sonophoresis of sali-
an ultrasound beam is incident upon it from either cylic acid and lanthanum tracers across hairless rat
side. Hence occurrence of air bubbles should be mini- skin in vivo using high-frequency ultrasound (f ¼ 2,
mized in the coupling medium in order to avoid ultra- 10, and 16 MHz) (Table 3). They investigated the
sound reflection. The reflection coefficient for various
Trace–Ultrasonic

dependence of sonophoresis on ultrasound frequency

interfaces may be estimated from the acoustic in the high-frequency region and found that 10 MHz
impedances of two media forming the interface using ultrasound is more effective in enhancing transdermal
equations described in Refs.[57,58]. transport of salicylic acid than that at 16 MHz, which
in turn, is more effective than that at 2 MHz. They
proposed that the sonophoretic enhancement in
Selection of Ultrasound Parameters the high-frequency region should vary directly with
ultrasound frequency, though the anomolusly high
Proper selection of ultrasound parameters is required efficiency of sonophoresis at 10 MHz was due to
to ensure safe and efficacious sonophoresis. Ultra- higher efficiency of the transducer operating at that
sound parameters such as frequency, intensity, duty frequency.
cycle, and distance of transducer from the skin influ- Low-frequency ultrasound (below 1 MHz): Tachibana
ence the efficiency of sonophoresis. Below we present et al.[52,53] have reported use of low-frequency ultra-
a general discussion of the role played by various ultra- sound (48 kHz) to enhance transdermal transport of
sound parameters in sonophoresis. Note that the lidocaine and insulin across hairless mice skin. Very
objective of this discussion is not to point out the low-frequency ultrasound has also been used by
exact values of ultrasound parameters to be selected, Mitragotri, Blankschtein, and Langer[37,38] to enhance
but rather to present information regarding the transport of various low-molecular weight drugs
dependence of sonophoretic enhancement on each including salicylic acid, corticosterone as well as
parameter. high-molecular weight proteins including insulin,
g-interferon, and erythropoeitin across human cadaver
Ultrasound frequency skin in vitro. They investigated the dependence of sono-
phoretic enhancement in low-frequency region using
Ultrasound at various frequencies in the range of two ultrasound frequencies, 20 and 40 kHz, and found
20 kHz to 16 MHz has been used for sonophoresis. that the sonophoretic enhancement of transdermal
These studies of sonophoresis can be classified into salicylic acid flux induced by 20 kHz ultrasound is up
three categories based on the ultrasound frequency to 7-fold higher than that induced by 40 kHz ultra-
used, i.e., therapeutic, high-frequency, and low- sound at the same intensity. The inverse dependence of
frequency ultrasound. sonophoretic enhancement on ultrasound frequency
Table 2 Literature reports of therapeutic sonophoresis
Drug Molecular weight (Da) Experimental system Ultrasound conditions Experimental conclusionsa Ref.
2
Transdermal Delivery: Sonophoresis

Caffeine 194 Human skin in vitro 1 MHz, 2 W/cm 0.2  0.4 [37]
Hairless rat in vitro 1 MHz, 2 W/cm2 1 [33]
Corticosterone 346 Human skin in vitro 1 MHz, 2 W/cm2 3  0.6 [37]
2
Dexomethasone 392 Swine 1 MHz, 1.5 W/cm Significant enhancement
Estradiol 272 Human skin in vitro 1 MHz, 2 W/cm2 12  1.5 [37]
Fluocinolone acetonide 452 Human skin in vivo 1 MHz, 2 W/cm2 Significant enhancement [35]
Dogs 1 MHz, 0.3–1 W/cm2 Significant enhancement [42]
Human skin in vivo 1 MHz, up to 2 W/cm2 Significant enhancement [42]
Hydrocortisone 362 Human skin in vivo 1 MHz, up to 3 W/cm2 Significant enhancement [42]
Human skin in vivo 1 MHz, 1.5 W/cm2 Significant enhancement [42]
Swine 1 MHz, 1.5 W/cm2 Significant enhancement [42]
Pigs 1 MHz, up to 3 W/cm2 Significant enhancement [42]
Indomethacin 357 Rats 1 MHz, 0.75 W/cm2 Significant enhancement [40]
Human skin in vitro 1 MHz, 2 W/cm2 0.1  0.6 —b
Lidocaine 234 Human skin in vivo 1 MHz, 0.25 W/cm2 No enhancement [8]
Human skin in vivo 1–3 MHz, 1.5 W/cm2 No enhancement [53]
Phenylbutazone 308 Human skin in vivo 1 MHz, 2 W/cm2 Significant enhancement [13]
Physostigmine 275 Hairless rats in vivo 1 MHz, 3 W/cm2 Significant enhancement [32]
2
Progesterone 274 Human skin in vitro 1 MHz, 2 W/cm 0.1  0.5 [37]
Salicylate 138 Human skin in vivo 1 MHz, 1.5 W/cm2 No significant enhancement [15]
Human skin in vivo 1 MHz, 1.5 W/cm2 No significant enhancement [14]
Testosterone 288 Human skin in vitro 1 MHz, 2 W/cm2 4  1.1 [37]
a
The experimental conclusions are reported either as statistically significant or insignificant enhancement or in terms of a quantitative ratio of sonophoretic and passive skin permeability.
b
Unpublished data by J. Kost and R. Langer.
3831

Trace–Ultrasonic
 3832 Transdermal Delivery: Sonophoresis

Table 3 Literature reports of high frequency sonophoresis

Molecular Experimental Ultrasound Experimental
Drug weight (Da) system conditions conclusionsa Ref.
Salicylic acid 138 Hairless rat 2, 10, and 16 MHz 2–4 fold [11]
in vivo 200 mW/cm2 enhancement
Lanthanum tracers — Hairless rat 10 and 16 MHz Significant [12]
in vivo 200 mW/cm2 enhancement
a
The experimental conclusions are reported as statistically significant or insignificant enhancement.

was hypothesized to occur due to inverse dependence of vary significantly with the pulse length. For example,
cavitational effects on ultrasound frequency.[59] the cavitation threshold in an aqueous solution at
1 MHz changes from approximately 0.3 W/cm2[60] to
Ultrasound intensity 33 W/cm2[61] as the mode of ultrasound application
changes from continuous to pulsed, with a pulse length
Various ultrasound intensities in the range of of 1 ms applied every 10 ms. This is because under
0.1–2 W/cm2 have been used for sonophoresis. In most pulsed ultrasound, during the intervals between pulses,
cases, use of higher ultrasound intensities is limited by gas nuclei formed during the previous pulse have time
thermal effects. Several investigations have been per- to dissolve back into solution, and therefore, making it
formed to assess the dependence of sonophoretic more difficult to cavitate the solution.[62] Mitragotri et
enhancement on ultrasound intensity. Miyazaki, al.[36] reported that while a continuous application of
Mizuoka, and Takada.[41] found a relationship between therapeutic ultrasound (1 MHz, 2 W/cm2) increased
the plasma concentrations of indomethacin transported human skin permeability to estradiol by 13-fold, a
across the hairless rat skin by sonophoresis (therapeutic pulsed application (2 ms pulses applied every 10 ms)
conditions) and the ultrasound intensity used for this did not significantly enhance transdermal estradiol
flux. In very low-frequency ultrasound region, Kost[28]
Trace–Ultrasonic

purpose. Specifically, the plasma indomethacin concen-

tration at the end of three hours after sonophoresis reported that urea permeability of cuprophane mem-
(0.25 W/cm2) was about 3-fold higher than controls at branes increased from 6 to 56% as the ultrasound
the same time. However, increasing intensity by 3-fold (20 kHz) pulse length increased from 100 to 400 ms
(to 0.75 W/cm2) further increased sonophoretic enhance- (applied every second).
ment only by 33%. Mortimer, Trollope, and Roy[42]
found that application of ultrasound at 1 W/cm2 Distance of the transducer from the skin
increased transdermal oxygen transport by 40% while
that at 1.5 W/cm2 and 2 W/cm2 induced an enhancement The ultrasound pressure (or intensity) field around a
by 50% and 55%, respectively. transducer is quite complex. The intensity of the ultra-
In the very low-frequency ultrasound region sound passes through a series of maxima and minima
(20 kHz), Mitragotri, Blankschtein, and Langer[37] in a region near the transducer and beyond a certain
have reported that permeability of human skin in vitro distance, decreases monotonically with distance. The
to insulin increased by more than 100-fold as the ultra- region in which the ultrasound intensity passes through
sound intensity increased from 12.5 to 125 mW/cm2. the series of minima and maxima is referred to as the
This variation of sonophoretic skin permeability with near field, and the region beyond the near field is
ultrasound intensity is quite different from that observed referred to as the far field. The length of the near field
in therapeutic frequency region described above. of a transducer having an area of 1 cm2 operating at
1 MHz is 1.66 cm.[57] In most of the experiments
Pulse length reported in the literature, for which ultrasound
frequencies of 1 MHz or above have been used, the
Ultrasound can be applied either in a continuous or a distance of the skin from the transducer was probably
pulsed mode. A pulsed mode of ultrasound application less than 1.66 cm. As a result, in most reported experi-
is used many times because it reduces the severity of ments, the skin was in the near-field region. In very
adverse side effects of ultrasound, such as thermal low-frequency ultrasound region, Julian and Zenter[23]
effects. However, pulsed application of ultrasound studied the effect of transducer distance on the sono-
may have a significant effect on the efficacy of sono- phoretic permeability of benzoic acid through a poly-
phoresis. As will be discussed later, cavitational dimethylsiloxane membrane under low-frequency
effects, which play a crucial role in sonophoresis, conditions. They observed that the effect of 20 kHz
Transdermal Delivery: Sonophoresis 3833

ultrasound on the permeability of the membrane is intended for topical delivery of various drugs. Among
insensitive to the distance of the transducer from the all the drugs that have been used for sonophoresis,
membrane. This probably occurs because of the suc- much attention has been focused on anti-inflammatory
cessive reflections of ultrasound waves in the diffusion drugs. These include steroidal drugs such as hydro-
cell which prevents any systematic pressure pattern cortisone and dexamethasone and non-steroidal drugs
form forming in the diffusion cell. such as indomethacin and salicylate. Sonophoresis of
anti-inflammatory drugs offers an advantage over their
Ultrasound energy dose passive topical delivery in that ultrasound may deliver
drugs deeper into tissues. This is especially advan-
In a recent systematic study of the dependence of tageous in the case of delivery of anti-inflammatory
20 kHz sonophoresis on ultrasound parameters, drugs to muscles which lie deeper into the body. Griffin
Mitragotri et al. showed that the enhancement of skin and Touchstone.[18] reported that application of ultra-
permeability varies linearly with ultrasound intensity sound (1 MHz, 2 W/cm2) delivered hydrocortisone
and ultrasound on-time (for pulsed ultrasound, ultra- about 5 cm deep into pig tissues. This characteristic
sound on-time equals the product of total ultrasound property of sonophoresis has been used effectively by
application time and duty cycle), while is independent these investigators to deliver hydrocortisone to joints
of the ultrasound duty cycle. Based on those findings, for the treatment of rheumatoid arthritis.
the authors reported that there is a threshold energy The most commonly used technique of sonophoresis
dose for ultrasound induced transdermal drug trans- in these studies was to apply hydrocortisone in the
port. Once the threshold value is crossed, the enhance- form of an ointment on the skin and then apply ultra-
ment of skin permeability varies linearly with the sound by keeping the transducer in contact with the
ultrasound energy dose (J/cm2), which is calculated ointment. In some cases, the transducer was moved
as the product of ultrasound intensity and ultrasound in circular patterns to avoid a continuous exposure of a
on-time. This result indicates that ultrasound energy certain part of the skin to ultrasound. Although these
dose can be used as a predictor of the effect of studies were performed using different animal models,
20 kHz sonophoresis. The authors also indicated that application techniques, hydrocortisone concentrations
it is important to determine the threshold energy dose in the ointment, and exposure time, a measurable

Trace–Ultrasonic
for each individual sonophoresis system, for example, enhancement of hydrocortisone transport was reported
the real in vivo situation, because it may vary from in almost all cases. In contrast, most of the attempts to
system to system. Specifically, it may vary between enhance transdermal transport of lidocaine and salicy-
different skin models, as well as with the ultrasound lates have been less successful. In the case of lidocaine,
frequency and the distance of the transducer from the sonophoretic enhancement was measured in terms
the skin surface, etc. of reduction of onset time for anesthesia or prolonging
duration of anesthesia. In most cases, no significant
effect of ultrasound application on either induction
PREVIOUS STUDIES OF SONOPHORESIS time or duration of anesthesia has been reported.[55]
Similarly sonophoresis of salicylates from ointments
Numerous attempts of sonophoresis have been per- has not been found to induce any significant increase
formed over the last 40 years. As described earlier, in plasma salicylate levels.[14]
these attempts can be classified into three categories: Literature data reported in Table 2 indicate that
therapeutic frequency, high-frequency and low-frequency except in the case of steroids including hydrocortisone,
ultrasound. dexomethasone, testosterone, estradiol, and cortico-
sterone, application of therapeutic ultrasound results
in either minor or no enhancement of transdermal drug
Therapeutic Frequency Sonophoresis transport. Mitragotri et al.[36] presented a hypothesis
for this variation of sonophoretic enhancement from
The therapeutic ultrasound conditions correspond to a drug to drug based on their mechanistic conclusion
frequency in the range of 1–3 MHz and an intensity in that ultrasound induces disorganization of the SC lipid
the range of 0–2 W/cm2. Therapeutic ultrasound has been bilayers, thus increasing drug diffusivity and hence
attempted to enhance transdermal transport of more than permeability of the SC. This mechanism suggests that
15 drugs,[7–10,14,15,18–22,25,27,28,30–33,36,42–45,47–50,55,56,63] a drugs such as steroids which possess low passive
summary of which is provided in Table 2. diffusion coefficients through the SC bilayers[36] com-
Historically, the transdermal route of drug adminis- pared to those through the disordered SC bilayers
tration has been considered for topical rather than should be significantly enhanced by ultrasound
systemic delivery of drugs. Accordingly, most of the application. On the other hand, drugs such as lidocaine
sonophoresis experiments reported in Table 2 were and salicylic acid possessing a passive diffusion
 3834 Transdermal Delivery: Sonophoresis

coefficient comparable to that through the disorgan- ultrasound at even lower frequencies (20 kHz)
ized bilayer phase[36] may not be significantly enhanced enhances transdermal transport of various low-
by ultrasound application. A mathematical equation molecular weight drugs including corticosterone, and
was developed[39] to predict a priori whether appli- salicylic acid as well as high-molecular weight proteins
cation of therapeutic ultrasound under a typical such as insulin, g-interferon, and erythropoeitin across
condition, that is, 1 MHz, 2 W/cm2 will enhance trans- the human skin in vitro[37] Table 4 provides a summary
dermal transport of a given drug: of the literature reports of low-frequency sonophoresis.
Transdermal transport enhancement induced by
0:75 low-frequency ultrasound has been found to be much
Ko=w
E  more significant than that induced by therapeutic
4
104 P
ultrasound. For example, although application of
where E is sonophoretic enhancement of skin per- therapeutic ultrasound has no effect on transdermal
meability (dimensionless), Ko/w is the octanol–water permeation of lidocaine, low-frequency ultrasound has
partition coefficient, and P is the passive skin perme- been shown to significantly enhance lidocaine trans-
ability (cm/h). A list of Ko/w and P values for various port across hairless rat skin in vivo. Quantitatively,
drugs may be found in Refs.[64,65] respectively. Mitragotri, Blankschtein, and Langer[38] compared
Over the last 20 years, transdermal route of delivery the enhancement ratios (ratio of the sonophoretic
has been considered as a means for systemic drug and passive permeabilities measured in vitro across
administration. Over this period, sonophoresis has human cadaver skin) induced by therapeutic ultra-
been attempted to enhance systemic transdermal deliv- sound (1 MHz, 2 W/cm2, continuous)[36] and low-
ery. Levy et al.[30] showed that 3–5 min of ultrasound frequency ultrasound (20 kHz, 125 mW/cm2, 100 msec
exposure (1 MHz, 1.5 W/cm2) increased transdermal pulses applied every second) in the case of four per-
permeation of mannitol and physostigmine across meants, butanol, corticosterone, salicylic acid, and
hairless rat skin in vivo by up to 15-fold. They also sucrose. They found that the enhancement induced
reported that the lag time typically associated with by low-frequency ultrasound is up to 1000-fold higher
transdermal drug delivery was nearly-completely elimi- than that induced by therapeutic ultrasound.[38]
nated after exposure to ultrasound.[30] Low-frequency ultrasound has been found to
Trace–Ultrasonic

Although several attempts have been made to enhance transdermal transport of drugs which do not
enhance transdermal drug transport using therapeutic permeate skinpassively, for example, large-molecular
ultrasound, a typical enhancement induced by thera- weightproteins. Application of low-frequency ultra-
peutic ultrasound is about 10-fold or smaller. This sound (20 kHz, 125 mW/cm2, 100 ms pulses applied
enhancement, may be sufficient for local delivery of every second) has been shown to enhance transdermal
certain drugs such as hydrocortisone, but is not suf- transport of proteins including insulin, g-interferon,
ficient for the systemic delivery of many drugs. Accord- and erythropoeitin across human cadaver skin in
ingly, despite of significant attention dedicated to vitro.[37] Figure 2 shows the variation of in vitro human
sonophoresis, there is no commercially available sono- skin permeability to insulin with ultrasound (20 kHz,
phoresis system for the systemic drug delivery. 100 ms pulses applied every second) intensity. Skin
insulin permeability increases by more than 100-fold as
the ultrasound intensity increases from 12.5 mW/cm2
Low-Frequency Sonophoresis to 125 mW/cm2. It has also been shown that appli-
cation of ultrasound under conditions the same as
Less attention has been given to sonophoresis in those mentioned above delivers therapeutic doses of
this ultrasound region (Table 4). Tachibana and insulin across hairless rat skin in vivo from a chamber
Tachibana[51–53] reported that application of low- glued on the rats back and filled with an insulin solu-
frequencyultrasound (48 kHz) enhances transdermal tion (100 U/ml).[37] Figure 3 shows blood glucose levels
transport oflidocaine and insulin across hairless rat of diabetic rats during ultrasound-insulin treatment.
skin in vivo. They found that the blood glucose level Insulin-ultrasound treatment (20 kHz, 225 mW/cm2,
of a hairless rat immersed in a beaker filled with insulin 100 ms pulses applied every second) reduces the blood
solution (20 U/ml) and placed in an ultrasound bath glucose level of diabetic hairless rats from about 400–
(48 kHz, 5000 Pa or 37 mm Hg) decreased by 50% in 200 mg/dl (the blood glucose level of normal rats) in
240 min.[51] They also showed that application of 30 min. A corresponding change in the plasma insulin
ultrasound under similar conditions prolongs the anes- levels was observed during sonophoresis. Normal hair-
thetic effect of transdermally administrated lidocaine less rats possessed a plasma insulin level of 101  31
in hairless rats[53] and enhances transdermal transport picomolar while diabetic hairless rats possessed a value
of insulin in rabbits.[52] Mitragotri, Blankschtein, below our detection limit (34 picomolar). During sono-
and Langer[37,38] have shown that application of phoresis, the levels of transdermally delivered human
Transdermal Delivery: Sonophoresis 3835

Table 4 Literature reports of low frequency sonophoresis

Molecular Experimental Ultrasound Experimental
Drug weight (Da) system conditions conclusionsa Ref.
Corticosterone 346 Human skin 20 kHz, 80 [38]
in vitro 125 mW/cm2
Estradiol 272 Human skin 20 kHz, 3 [38]
in vitro 125 mW/cm2
Salicyclic acid 138 Human skin 20 kHz, 400 [38]
in vitro 125 mW/cm2
Aldosterone 360 Hairless rat 20 kHz, 1400 [38]
in vitro 125 mW/cm2
Water 18 Human skin 20 kHz, 113 [38]
in vitro 125 mW/cm2
Lidocaine 234 Rat skin 48 kHz Significant [53]
in vivo enhancement
Insulin 6000 Human skin 20 kHz, Significant [52]
in vitro up to 225 mW/cm2 enhancement
Rat skin
in vivo
Insulin 6000 Rat skin 48 kHz, Significant [37]
enhancement
g-Interferon 17,000 Human skin 20 kHz, Significant [37]
in vitro up to 225 mW/cm2 enhancement
Rat skin
in vivo
Erythropoeitin 48,000 Human skin 20 kHz, Significant [37]

Trace–Ultrasonic
in vitro up to 225 mW/cm2 enhancement
Rat skin
in vivo
a
The experimental conclusions are reported either as statistically significant or insignificant enhancement or in terms of a quantitative ratio of
sonophoretic and passive skin permeability.

insulin in rat plasma reached a value of 77 (28) pico- 20 minute application of ultrasound (0.2 W/cm2) at
molar after 30 min. and a value of 178 (84) picomolar a frequency of 2 MHz did not significantly enhance
after 1 h.[37] No significant change in the plasma amount of salicylic acid penetrating the skin. However,
concentration of indigenous rat insulin was observed 10 MHz ultrasound under otherwise same conditions
during sonophoresis. These results indicate that sono- resulted in about a 4-fold increase and 16 MHz ultra-
phoresis at 20 kHz delivers therapeutic doses of insulin sound resulted in about a 2.5-fold increase in transder-
across hairless rat skin in vivo. mal salicylic acid transport.[11] They also investigated
the effect of shorter (5 min) ultrasound exposures
under similar conditions on transdermal salicylic acid
High-Frequency Sonophoresis transport and found that while 10 MHz ultrasound
enhances transdermal transport by 1.6-fold, that at
This region of ultrasound corresponds to a frequency 16 MHz enhances it by about 1.8-fold. Application of
higher than 3 MHz. Bommanan et al.[11] hypothesized high-frequency ultrasound was also found to reduce
that since the absorption coefficient of the skin varies the long lag time associated with transdermal trans-
directly with the ultrasound frequency, high frequency port. Bommanan et al.[11] found that transdermally
ultrasound energy would concentrate more in the delivered salicylic acid appears much sooner in the
epidermis, thus leading to higher enhancements. In urine if driven by sonophoresis than by passive per-
order to assess this hypothesis, they studied the effect meation. These researchers also found that an electron
of high-frequency ultrasound (2–15 MHz) on the per- dense tracer, such as lanthanum, was driven deep into
meability of salicylic acid (dissolved in a gel) through the dermis by a 5 min application of high-frequency
hairless guinea pig skin in vivo. They found that a ultrasound in hairless mouse in vivo.
 3836 Transdermal Delivery: Sonophoresis

10–2
a-values for several biological tissues can be found in
Refs.[60,66] (Table 1). The absorption coefficient of a
Skin Insulin Permeability (cm/hr)

medium increases proportionally with the ultrasound

10–3 frequency indicating that, the thermal effects of ultra-
sound are proportional to the ultrasound frequency.
The increase in the temperature of a medium upon
10–4 ultrasound exposure at a given frequency varies pro-
portionally with the ultrasound intensity and exposure
time. The thermal effects can be substantially decreased
10–5 by pulsed application. For a detailed discussion of the
thermal effects of ultrasound.[60]

10–6
0 50 100 150 200 250 Acoustic streaming
Ultrasound Intensity (mW/cm2)
Acoustic streaming, by definition, is the development
Fig. 2 Human skin permeability to insulin. (From Ref.[37].) of time independent large fluid velocities in a medium
under the influence of an ultrasound wave. The pri-
MECHANISMS OF SONOPHORESIS mary causes of acoustic streaming are the reflections
and other distortions of the wave propagation. Oscilla-
In order to understand the mechanisms of sonophoresis, tions of cavitation bubbles may also contribute to
it is important to identify various effects of ultrasound acoustic streaming. The shear stresses developed
exposure on the human tissue since one or more of by streaming velocities may affect the neighboring
these effects may contribute to the mechanism of structures.[67]
sonophoresis. A brief description of the various biologi-
cal effects of ultrasound is provided below. Cavitational effects

Cavitation is the formation of gaseous cavities in a

Trace–Ultrasonic

medium upon ultrasound exposure. The primary cause

Exposure Effects of cavitation is the ultrasound-induced pressure varia-
tions in the medium. Cavitation involves either rapid
Thermal effects growth and collapse of a bubble (transient cavitation)
or slow oscillatory motion of a bubble in ultrasound
Absorption of ultrasound results in a temperature field (stable cavitation). Cavitation affects tissues in
increase of the medium. Materials which posses higher several ways. Specifically, collapse of cavitation bub-
ultrasound absorption coefficients, such as bones, bles releases a shock wave which can cause a structural
experience severe thermal effects as compared to muscle alterations in its surroundings. Biological tissues
tissues which have a lower absorption coefficient (a). contain numerous air pockets trapped in the fibrous
structures which act as nuclei for cavitation upon
500 ultrasound exposure. Accordingly, a significant cavi-
tation activity is known to occur in biological tissues
Blood Glucose Level (mg/dl)

450 upon ultrasound exposure.[60] The cavitational effects

400 vary inversely with ultrasound frequency and directly
with ultrasound intensity. Significant attention has
350
been devoted to explore which of the above mentioned
300 phenomena plays an important role in sonophoresis.
250

200 Mechanisms of Therapeutic Sonophoresis

150 Us on Us off Mortimer, Trollope, and Roy[42] performed sonophor-

100 esis of oxygen across frog skin in vitro. They found
0 0.5 1 1.5 2
that the sonophoretic enhancement of transdermal
Time (hour)
oxygen transport depends on ultrasound intensity,
Fig. 3 Blood glucose levels of rats; , diabetic hairless rats; rather than pressure amplitude. Based on this obser-
`, normal rats, G, diabetic rats treated by sonophoresis and vation, they hypothesized that cavitation cannot be
insulin for 30 min. (From Ref.[37].) responsible for sonophoresis. They hypothesized that
Transdermal Delivery: Sonophoresis 3837

the observed enhancement occurs due to acoustic degassed skin was exposed to ultrasound, once again,
streaming in the solution around the skin.[42] Levy the effect of ultrasound on the estradiol permeability
et al.[30] performed an in vitro investigation of the roles was minimal (1.5-fold), compared to 13-fold across
played by thermal effects, cavitation, and mixing in the normal skin. Based on these two results, the
sonophoretic enhancement of urea transport across authors concluded that cavitation inside the skin plays
polymer membranes. They found that the observed a major role in enhancing transdermal transport
enhancement cannot be explained by the thermal upon therapeutic ultrasound exposure. They provided
effects or mixing. In an attempt to elucidate the role the following hypothesis for the mechanism of sono-
played by cavitation, they performed sonophoresis phoresis performed using therapeutic ultrasound.
experiments using degassed solutions. Since degassing Ultrasound exposure in the therapeutic range causes
a solution decreases the cavitation activity in the solu- cavitation in the keratinocytes of the stratum corneum.
tion, they hypothesized that if a decrease in the sono- Oscillations of the ultrasound-induced cavitation
phoretic enhancement is observed upon degassing, it bubbles near the keratinocyte-lipid bilayer interfaces
would indicate the importance of cavitation. Indeed, may, in turn, cause oscillations in the lipid bilayers,
they found that degassing procedure reduced the sono- thereby causing structural disorder of the SC lipids
phoretic enhancement of urea permeation by 2-fold sug- (Fig. 4). Shock waves generated by the collapse of cavi-
gesting that cavitation may play a role in sonophoresis. tation bubbles at the interfaces may also contribute to
Cavitation occurs in a variety of mammalian tissues, the structure-disordering effect.
including muscle, abdominal tissues, brain, cardio- Since diffusion of permeants through a disordered
vascular tissues, and liver upon exposure to ultrasound bilayer phase can be significantly higher than that
at a variety of conditions.[60] As explained earlier, the through a normal bilayer, transdermal transport in
occurrence of cavitation in biological tissues is attribu- the presence of ultrasound is expected to be higher
ted to the existence of a large number of gas nuclei. than passive transport.
These nuclei are gas pockets trapped in either intra-
cellular or intercellular structures. Simonin et al.[68]
hypothesized that cavitation occurs in the follicles of Mechanisms of Low-Frequency
the skin upon ultrasound exposure and enhances trans- Sonophoresis

Trace–Ultrasonic
dermal permeation by convective velocities through
follicles. However, no evidence was presented to Since cavitational effects in fluids vary inversely with
support this hypothesis. Mitragotri et al.[36] presnted ultrasound frequency,[59] it is likely that cavitational
results of the experiments indicating that cavitation effects should play an even more important role in
inside the skin plays an important role in sonophoresis low-frequency sonophoresis. Tachibana et al.[53] hypo-
performed using therapeutic ultrasound. thesized that application of low-frequency ultrasound
In the first set of experiments, the known effect of results into acoustic streaming in the hair follicles
static pressure on cavitation was utilized. It is known and sweat ducts of the skin, thus leading to enhanced
that cavitation in fluids and porous media[21] can be transdermal transport. Mitragotri et al.[38] hypohesized
suppressed at high pressures. This effect is believed to that transdermal transport during low-frequency sono-
occur due to the dissolution or collapse of the gaseous phoresis occurs across the keratinocytes rather than
nuclei under the influence of pressure. Sonophoresis hair follicles. They provided the following hypothesis
experiments were performed using skin compressed at for the higher efficacy of low-frequency sonophoresis.
30 atm (between two smooth glass plates soaked in Cavitation induced by low-frequency ultrasound
water placed in a compression press for two hours may cause disordering of the SC lipids. In addition,
prior to sonophoresis experiments). They found that oscillations of cavitation bubbles may result in signi-
while application of ultrasound (1 MHz, 2 W/cm2, ficant water penetration into the disordered lipid
continuous) enhances estradiol permeability of the regions. This may cause the formation of aqueous
normal human epidermis by 13-fold, the correspond- channels through the intercellular lipids of the SC
ing enhancement for compressed skin is only about through which permeants may transport (Fig. 5). The
1.75-fold. occurrence of transdermal transport through aqueous
In the second set of experiments, the heat-stripped channels across the disordered lipid regions may
human cadaver skin was degassed (under a pressure enhance transdermal transport as compared to passive
of 0.05 mm Hg) prior to the permeability experiments. transport because i) the diffusion coefficients of per-
The authors hypothesized that when a skin piece meants through water, which is likely to primarily
soaked in buffer is subjected to high vacuum, the occupy the channels generated by ultrasound, are up
resulting low pressures should reduce the dissolved to 1000-fold higher than those through the ordered
gas concentration in the buffer thereby forcing small lipid bilayers[38] and ii) the transport path length of
gaseous nuclei in the skin to dissolve. When the these aqueous channels may be much shorter (by a
 3838 Transdermal Delivery: Sonophoresis

Fig. 4 Ultrasound-induced cavitation bubbles in stratum corneum.

factor up to 25[69]) than that though the tortuous inter- drugs through water is much faster than that through
cellular lipids in the case of passive transport. ordered lipid bilayer regions, thus allowing drugs to
This hypothesis also explains why low-frequency transport across the skin at a faster rate. Therefore,
ultrasound can induce transdermal transport of drugs molecules such as hydrophilic drugs or proteins, may
which exhibit very low passive transport. Drugs permeate skin with relative ease in the presence of
Trace–Ultrasonic

possessing low passive permeabilities are either i) low-frequency ultrasound.

hydrophilic, which makes their partitioning into the
SC bilayers difficult or ii) large in molecular size (for
example, proteins), which reduces their diffusion coef- Mechanisms of High-Frequency
ficients in the SC. Low-frequency ultrasound may Sonophoresis
overcome both of these limitations by providing aque-
ous transport channels across the skin. Since these Bommanan et al.[12] perfored sonophoresis of lantha-
channels are filled with saline, hydrophilic drugs can num tracers across hairless mice skin at an ultrasound
easily partition into the SC. In addition, diffusion of frequency of 16 MHz in order to understand the

Fig. 5 Transdermal transport through the stratum corneum. (A) Passive and (B) during low-frequency sonophoresis.
Transdermal Delivery: Sonophoresis 3839

transport pathways during high-frequency sono- application of low-frequency ultrasound results in

phoresis. They observed the skin under the electron anypermanent loss of the barrier properties of skin
microscope after sonophoresis and found that 5 min measured interms of water permeability.[38] It has been
of sonophoresis results in penetration of lanthanum found that in the case of a 1 h long ultrasound
tracers to dermal levels of the skin. They further exposure, the skin permeability to water measured
reported that the tracer was patchily distributed within 2 h post-exposure was comparable to the passive
within the intercellular lipid bilayers of the SC. They skin permeability to water. In the case of a 5-h long
provided the following hypothesis for the mechanism ultrasound exposure, the skin permeability 2 h post-
of high-frequency sonophoresis. The micronuclei (air- exposure was about 6 times higher than the passive
pockets) present in the SC oscillate in response to permeability to water. However, this value continued
oscillating pressure fields of ultrasound and eventually to decrease, and was within a factor of 2 of the passive
collapse. The oscillations of these bubbles result in skin water permeability 12 h post-exposure. Studies
enhanced skin permeation. They also hypothesized have also been performed[35] to assess whether
that the patchy distribution of the lanthanum tracer application of high-frequency ultrasound induces
revealed in the micrographs corresponds to the any irreversible damage to the barrier properties of
location of oscillating air pockets in the SC. In a later the skin measured in terms of trans-epidermal water
report, Menon, Bommanon, and Elias[35] presented loss (TEWL) across hairless mice skin exposed to
additional microscopic studies of the hairless mice high-frequency ultrasound (16 MHz). Nosignificant
skin after undergoing sonophoresis of lanthanum difference in TEWL values of the skin exposed to
tracer. They reported the presence of long confluent ultrasound and that not exposed to ultrasound was
channels in the intercellular lipids filled with lantha- found.[35]
num tracers in the hairless rat skin exposed to
ultrasound. They presented the following hypothesis
for the mechanism of sonophoresis: application
of ultrasound opens and expands gas-filled cavities Biological Effects of Low-Frequency
in the SC much like pumping air through a collapsed Ultrasound
rubber tubing. Enhanced transport of drugs may
Ultrasound over a wide frequency range has been used

Trace–Ultrasonic
then occur through these confluent channels across
the SC. in medicine over last century. For example, therapeutic
ultrasound (1–3 MHz) has been used for massage,
low-frequency ultrasound has been used in dentistry
(23–40 kHz),[70,71] and high-frequency ultrasound
SAFETY
(3–10 MHz) has been used for diagnostic purposes.[66]
In view of this, significant attention has been dedicated
The safety aspects of sonophoresis involve the revers-
to investigate the effects of ultrasound on biological
ibility of the skin barrier properties after turning ultra-
tissues. However, no conclusions have been reached
sound off, and the effect of ultrasound on the living
regarding the limiting ultrasound conditions required
parts of the skin and underlying tissues. Many reports to ensure safe exposure.
exist in literature describing preliminary assessments of
As described earlier, ultrasound affects biological
sonophoresis with respect to these two issues.
tissues via three main effects, thermal effects, cavitational
effects, and acoustic streaming. Conditions under which
these effects become critical are given below.[60]
Recovery of the Skin Barrier Properties Thermal effects may be important when
After Sonophoresis
1. The tissue has a high protein content.
Numerous reports exist to suggest that application of 2. A high intensity continuous wave ultrasound is
therapeutic ultrasound (1–3 MHz, 0–2 W/cm2) does used.
not induce any irreversible change in the skin per- 3. Bone is included in the heated volume.
meability to drugs in vivo. Quantitative measurements 4. Vascularization is poor.
of estradiol transport across human skin (in vitro) have
also shown that application of therapeutic ultrasound Cavitation may be important when
(1 MHz, 2 W/cm2) does not induce any statistically
significant irreversible change in skin barrier proper- 1. Low-frequency ultrasound is used.
ties.[36] Similar studies have also been performed using 2. Gassy fluids are exposed.
very low-frequency ultrasound (20 kHz, 125 mW/cm2, 3. Small gas-filled spaces are exposed.
100 ms pulses applied every second) to assess whether 4. The tissue temperature is higher than normal.
 3840 Transdermal Delivery: Sonophoresis

Streaming may be important when critical for efficient sonophoresis. The numerous
attempts made over the last 50 years can be classified
1. The medium has an acoustic impedance differ- into three categories: therapeutic frequency, high-
ent from its surroundings. frequency and low-frequency ultrasound; the first repre-
2. The fluid in the biological medium is free to sents the most commonly used ultrasound condition
move. for sonophoresis, although recently, attention has been
3. Continuous wave application is used. more focused on low- and high-frequency conditions.
Mechanistic experiments performed by several investi-
Numerous investigators have performed histologi- gators suggest that cavitation plays a major role. It has
cal studies of animal and human skin exposed to ultra- been suggested that cavitation disorganizes the lipid
sound under various conditions in order to assess the bilayers of the skin through which enhanced transport
effect of ultrasound on living skin cells. Levy et al.[30] of drugs may occur. Various studies have indicated that
exposed in vivo hairless rat skin to therapeutic ultra- application of ultrasound under conditions used for
sound (1 MHz, 1.5 W/cm2 continuous wave or sonophoresis does not cause any permanent damage to
3 W/cm2 pulsed wave, 3–5 min) and reported no dam- the skin or underlying tissues, although more work is
age to the skin. Nevertheless, Machet et al.[72] reported required before arriving at definite conclusions regarding
both epidermal and dermal alteration of human and the safety of ultrasound exposure.
hairless mouse skin when exposed to therapeutic ultra-
sound in vitro (3.3 MHz, 3 W/cm2, 10 min). The
authors proposed that a combined cavitation and
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Trace–Ultrasonic

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it is used clinically. J. Am. Phys. Ther. Assoc. 1966, 46, 41. Miyzaki, S.; Mizuoka, O.; Takada, M. External control of
18–26. drug release and penetration: enhancement of the transder-
20. Griffin, J.E.; Echternach, J.L.; Proce, R.E.; Touchstone, mal absorption of indomethacin by ultrasound irradiation.
J.C. Patients treated with ultrasonic driven hydro- J. Pharm. Pharmacol. 1990, 43, 115–116.
cortisone and with ultrasound alone. Phys. Ther. 1967, 47, 42. Mortimer, A.J.; Trollope, B.J.; Roy, O.Z. Ultrasound-
600–601. enhanced diffusion through isolated frog skin. Ultrasonics
21. Griffin, J.E.; Touchstone, J.C. Low-intensity phono- 1988, 26, 348–351.
phoresis of cortisol in swine. Phys. Ther. 1968, 48, 1136– 43. Newman, J.T.; Nellermo, M.D.; Crnett, J.L. Hydrocorti-
1344. sone phonophoresis: a literature review. J. Am. Pod. Med.
22. Griffin, J.E.; Touchstone, J.C. Effects of ultrasonic fre- Assoc. 1992, 82, 432–435.
quency on phonophoresis of cortisol into swine tissues. 44. Novak, E.J. Experimental transmission of lidocaine
Am. J. Phys. Med. 1972, 51, 62–78. through intact skin by ultrasound. Arch. Phys. Med.
23. Julian, T.N.; Zentner, G. Ultrasonically mediated solute Rehab. 1964, May, 231–232.
permeation through polymer barriers. J. Pharm. Pharma- 45. Oziomek, R.S.; Perrin, D.H.; Herold, D.A.; Denegar, C.R.
col. 1986, 38, 871–877. Effect of phonophoresis on serum salicylate levels. Med.
24. Julian, T.N.; Zentener, G.M. Mechanim for ultrasonically Sci. in Sports and Exer. 1990, 23, 397–401.
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Rel. 1990, 12, 77–85. pedic Clinics of North America 1982, 13, 579–586.
25. Kleinkort, J.A.; Wood, F. Phonophoresis with 1 percent 47. Pottenger, J.F.; Karalfa, L.B. Utilization of hydrocortisone

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versus 10 percent hydrocortisone. Phys. Ther. 1975, 55, phonophoresis in united states army physical therapy
1320–1324. clinics. Milit. Med. 1989, 154, 355–358.
26. Kost, J.; Levy, D.; Langer, R. Ultrasound as a transdermal 48. Pratzel, H.; Ditrich, P.; Kukovetz, W. Spontaneous and
enhancer. In Percutaneous Absorption Mechanisms- forced cutaneous absorption of indomethacin in pigs and
Methodology-Drug Delivery; Bronaugh, R., Maibach, humans. J. Rheumat. 1986, 13, 1122–1125.
H.I., Eds.; Marcel Dekker, Inc.: New York, 1989; 49. Quillen, W.S. Phonophoresis: a review of the literature and
595–601. technique. Athelet. Train. 1980, 15, 109–110.
27. Kost, J.; Langer, R. Ultrasound-mediated transdermal drug 50. Skauen, D.M.; Zentner, G.M. Phonophoresis. Int. J.
delivery. In Topical Drug Bioavailability, Bioequivalence, Pharm. 1984, 20, 235–245.
and Penetration; Shah, V.P., Maibach, H.I.,, Eds.; 51. Tachibana, K.; Tachibana, S. Transdermal delivery of
Plennum: New York, 1993; 91–103. insulin by ultrasonic vibration. J. Pharm. Pharmacol.
28. Kost, J. Ultrasound for controlled delivery of therapeutics. 1991, 43, 270–271.
Clin. Mater. 1993, 13, 155–161. 52. Tachibana, K. Transdermal delivery of insulin to alloxan-
29. Lenart, I.; Auslander, D. The effects of ultrasound on dif- diabetic rabbits by ultrasound exposure. Pharm. Res.
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ultrasound on transdermal drug delivery to rats and guinea lidocaine. Anestheiology 1993, 78, 1091–1096.
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31. Machluf, M.; Kost, J. Ultrasonically enhanced transdermal electron microscopy evaluation of the skin surface after
drug delivery. Experimental approaches to elucidate the ultrasound exposure. Anat. Rec. 1997, 247, 455–461.
mechanism. J. Biomat. Sci. 1993, 5, 147–156. 55. Williams, A.R. Phonophoresis: an in vivo evaluation using
32. McElnay, J.C.; Matthews, M.P.; Harland, R.; McCafferty, three topical anaesthetic preparations. Ultrasonics 1990, 28,
D.F. The effect of ultrasound on the percutaneous absorp- 137–141.
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acetonide. Int. J. Pharm. 1987, 40, 105–110. Fundamentals of Acoustics; John Wiley & Sons: New
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Phonophoresis of methyl nicotinate: a preliminary study 58. Hueter, T.F.; Bolt, R.H. Sonics: Techniques for the Use of
to elucidate the mechanism. Pharm. Res. 1993, 4, 1726– Sound and Ultrasound in Engineering and Science; John
1731. Wiley & Sons: New York, 1962.
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130–139. logical Effects; VCH Publishers: New York, 1989.
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61. Crum, L.A.; Folwlkes, J.B. Acoustic cavitation generated 67. Nyborg, W.L. Acoustic streaming. In Acoustic Streaming;
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63. Tyle, P.; Agrawala, P. Drug delivery by phonophoresis. 69. Edwards, D.; Langer, R. A linear theory of transdermal
Pharm. Res. 1989, 6, 355–360. transport phenomena. J. Pharm. Sci. 1994, 83, 1315–
64. Hanch, C.; Leo, A. Substituent Constants for Correlation 1334.
Analysis in Chemistry and Biological Sciences; Wiley: 70. Walmsley, A.D. Applications of ultrasound in dentistry.
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Gerrity, T.R., Henry, C.J., Eds.; Elsevier: New York, 72. Machet, L.; Pinton, J.; Patat, F.; Arbeille, B.; Pourcelot, L.;
1990; 93–127. Vaillant, L. In vitro phonophoresis of digoxin across hair-
66. Wells, P.N.T. Biomedical Applications of Ultrasound; less mice and human skin: thermal effect of ultrasound.
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Trace–Ultrasonic
Transdermal Delivery: Technologies
S. Kevin Li
Department of Pharmaceutics and Pharmaceutical Chemistry, College of Pharmacy,
University of Utah, Salt Lake City, Utah, U.S.A.

INTRODUCTION granulosum, stratum spinosum, and stratum basale.

The dermis layers passes into the subcutaneous tissue
The advantages of transdermal drug delivery over layer without a sharp boundary and consists of mostly
conventional routes of drug delivery are many folds. connective tissues with thickness of approximately
Examples of approved transdermal applications are 1–2 mm. The dermis also consists of an extensive plexus
estradiol and testosterone in hormone replacement, of blood capillaries, lymphatics, and nerves. Hair folli-
clonidine for hypertension, fentanyl and lidocaine for cles and glands (sebaceous gland and eccrine sweat
pain management, nicotine for smoking cessation, glands) are the appendages in the skin. These appen-
nitroglycerin for angina pectoris, norelgestromin/ dages are formed by specialized cells developed in the
ethinyl estradiol for contraception, scopolamine for fetal life. The appendages extend down from tubular
motion sickness, and alprostadil for erectile dysfunc- opening of the epidermis into the dermis, where they
tion. Only a limited number of drugs can be success- are surrounded by epithelial cells similar to those of
fully developed into transdermal products because of the epidermis.
the skin barrier. To overcome the skin barrier, chemi- The stratum corneum is the outermost layer of the
cal and physical enhancers are used to facilitate drug epidermis and has a thickness of 10–15 mm. It is the
transport across the skin. A complete review of this principal barrier for the transport of most solutes
topic is exhaustive and will not be included in this (except for very lipophilic compounds) across the skin.

Trace–Ultrasonic
chapter. Among the physical enhancers, the concept of The stratum corneum is a continuous heterogeneous
using electric field to enhance transdermal transport has structure that consists of approximately 10–25 layers
been around for decades, but has only evolved into com- of closely packed dead keratinized cells (corneocytes)
mercial products for drug delivery in recent years. Recent cemented together by intercellular lipids. The inter-
technologies such as ultrasound, abrasion, and micronee- cellular lipids in the stratum corneum are in the form
dles are based on the mechanism of disrupting the skin of multiple lamellar bilayers composed mainly of
barrier with physical means. Although this chapter will ceramides, cholesterol, and fatty acids. Proteins in the
provide a general overview of transdermal drug delivery stratum corneum are largely concentrated within the
and technologies, the focus will be on technologies corneocytes as keratin fibrils. The transport of lipophi-
already used in devices in late stage clinical trials, recently lic compounds across the stratum corneum is related
completed NDA filing, or approved by FDA. Parti- to the intercellular lipids (lipoidal or intercellular
cularly, the mechanisms of enhanced transdermal trans- pathways).[1] On the other hand, it is believed that
port such as iontophoresis will be discussed. Recent the transport of polar and ionic compounds is related
advances and improvement in transdermal iontophoresis to pathways with aqueous properties (the polar or pore
will be reviewed. A brief overview will also be provided pathways) when the stratum corneum is under a
for other physical enhancers that facilitate transdermal hydrated state.[2–6]
transport such as electroporation, sonophoresis, heat,
and abrasion with microneedles, heat, and laser.
Transdermal Transport

Skin Morphology The transdermal route provides an alternative route to

oral administration. Drugs delivered by this route
Before the discussion of transdermal transport, it is avoid digestion in the gastrointestinal tract (GI), local
essential to understand the anatomy and barrier proper- metabolism related to the GI route, and hepatic first
ties of skin. Briefly, the skin can be viewed as two main pass metabolism. It can provide controlled drug
layers: the epidermis and the dermis. The thickness of delivery with the possibility of zero order (continuous)
epidermis varies from approximately 0.07 to 0.12 mm release. Thus, it is most suitable for short half-life
(except on the palms and the soles), and is composed therapeutic agents and drugs with narrow therapeutic
of the stratum corneum, stratum lucidum, stratum windows. The use of transdermal patches is more
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-120041572
Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved. 3843
 3844 Transdermal Delivery: Technologies

convenient than frequent oral dosing due to simplified For passive diffusion, the permeability coefficient of
regimens and easy procedure of self-administration. the lipoidal pathway can be expressed as:
This can improve patient compliance. The disadvan-
tages of transdermal drug delivery include local skin KL DL
P L ¼ eL ð4Þ
irritation and allergy, high local drug concentration hL
that may cause tissue toxicity at the site of application,
where eL is the porosity, KL the partition coefficient,
and local first-pass metabolism. There are also limita-
DL the diffusion coefficient, and hL the effective path-
tions in regard to the physicochemical properties
way length of the lipoidal pathway.
of the drugs that can utilize the transdermal route.
The permeability coefficient of the pore pathway
To illustrate this point, basic transport theory will be
can be described by:
discussed here.
The amount of drug transport across a membrane
eP DP
at steady-state under a sink condition in the receiver PP ¼ ð5Þ
can be expressed in terms of the flux (J) and per- hP
meability coefficient (P):
where eP is the porosity, DP the effective diffusion coef-
ficient, and hP the effective pathway length of the pore
Q ¼ JðAD tÞ ð1Þ pathway.
A more general equation for the transport of polar
J and ionic permeants across the pore pathway in the
P ¼ ð2Þ
CD membrane is:[9]


where CD is the concentration of the permeant in the dCP dc
J ¼ eP DP  u P CP  vP CP ð6Þ
donor chamber, AD is the diffusional surface area, Q dx dx
is the cumulative amount of the permeant transported
across the membrane, and t is the duration of where c is the electric potential, vP is the effective velo-
transport. city of convective solvent flow, CP is the concentration
Trace–Ultrasonic

For example, the permeability coefficients of intact of the permeant, x is the position of the permeant in
stratum corneum for lipophilic compounds such as the membrane, and uP is the effective electromobility
corticosterone are in the order of 2
107 cm/sec of the permeant.
in an aqueous system.[7] The aqueous solubility of Note that steep molecular size-dependent transport
corticosterone is approximately 0.3 mg/mL.[8] A simple of polar permeants across the stratum corneum has
calculation using Eqs. (1) and (2) show why only been observed due to the small effective pore size of
potent drug of a required dose less than 1 mg per day the transport pathways in the stratum corneum.[10]
can utilize the transdermal route via passive delivery. Therefore, the diffusion coefficient, velocity, and
Physical and chemical transdermal enhancers are electromobility expressed in Eqs. (5) and (6) are signifi-
required for most drugs. Table 1 provides a list of cantly smaller than the free aqueous diffusion and
transdermal enhancement methods and companies mobility values found in the literature due to hindered
involved in the development of technologies related transport. Such hindered transport effects have been
to these methods. The readers are encouraged to view studied previously [6,10] and will not be elaborated here.
the company websites listed in the table, but caution In general, the molecular size of a permeant should be
must be taken because the information and claims less than 800 Dalton for effective permeant transport
provided in a company website are subjective and across intact stratum corneum.
not peer-reviewed. According to Eqs. (1)–(6), the effects of chemical
To further illustrate the mechanisms of chemically and physical enhancers upon transdermal transport
and physically enhanced transdermal transport, a can be primarily divided into mechanisms such as
transdermal transport model will be discussed here. increasing the porosity of the stratum corneum (eP
Generally, the permeability of the stratum corneum and eL), increasing the effective pore size of the pore
can be divided into parallel lipoidal and pore pathway pathway (DP and vP), reducing the effective thickness
components:[8] of the membrane (hL and hP), enhancing the partition-
ing and diffusion of the permeant into and across the
PSC ¼ PL þ PP ð3Þ lipids in the membrane (KL and DL), and providing a
driving force such as an electric field (dc/dx) and con-
where PSC is the total permeability of the stratum cor- vention (vP). It should be noted that simply increasing
neum and PL and PP are the permeability coefficients drug solubility in a transdermal patch, for example by
of the lipoidal and pore pathways, respectively. cosolvents, does not significantly improve transdermal
Transdermal Delivery: Technologies 3845

Table 1 Summary of transdermal technologies

Category Mechanism Companya Address
Patch and Drug-in-adhesive, 3M Drug Delivery www.3m.com/us/healthcare/
material membranes, hydrogel, manufacturers/dds/
technology gel, adhesive, matrix Alza Corp www.alza.com
Antares Pharma www.antarespharma.com
Auxilium Pharmaceuticals www.auxilium.com
Berlex Laboratories www.berlex.com
LAM Pharmaceutical www.lampharm.com
Mylan Technologies www.mylantech.com
Noven Pharmaceuticals www.noven.com
Solvay Pharmaceuticals www.solvaypharmaceuticals-
us.com
Chemical Penetration 3M Drug Delivery www.3m.com/us/healthcare/
enhancer enhancer/retarder manufacturers/dds/
Adherex Technologies www.adherex.com
Alza Corp www.alza.com
Bentley Pharmaceuticals www.bentleypharm.com
CollaGenex Pharmaceuticals www.collagenex.com
MacroChem www.macrochem.com
NexMed www.nexmed.com
Carrier (microemulsion, 3M Drug Delivery www.3m.com/us/healthcare/
liposome, transfersome) manufacturers/dds/
IDEA AG www.idea-ag.de
Physical Electrotransport Iontophoresis See Table 2
enhancer Electroporation Genetronics Biomedical www.genetronics.com
Physical disruption Ultrasound Sontra Medical www.sontra.com
Heat Zars www.zars.com

Trace–Ultrasonic
Poration, abrasion, Altea Therapeutics www.alteatherapeutics.com
and microneedle Alza www.alza.com
Becton & Dickinson www.bd.com
Laser SpectRx www.spectrx.com
Norwood Abbey Ltd www.norwoodabbey.com
Pressure waves Norwood Abbey Ltd www.norwoodabbey.com
RF current TransPharma Medical www.transpharma-medical.
poration com
Needle-free injection Antares Pharma www.antarespharma.com
Chiron Vaccines www.powderject.com
a
There are more than a hundred pharmaceutical and biotech companies involved in some sorts of transdermal or skin research. This table is not
intended to provide a complete list of all these companies but a few examples to the readers. The omission of any companies in the list is not
intentional.

flux due to the resulting decrease in the partition reservoir patch systems. In this system (e.g., Estraderm
tendency of the drug from the patch into the stratum and Transderm Scop from Novartis, Androderm from
corneum (KL). An increase in the solubility, however, Watson Lab), drug is usually solubilized in a solvent
is necessary to increase the loading capacity of a such as alcohol in a reservoir with a rate-controlling
drug in the transdermal patch and reduce patch size membrane separating the drug reservoir and the skin.
(thickness). The adhesive that holds the patch on the skin is
not in direct contact with the diffusion surface area
but encircling it. In the drug-in-adhesive system (e.g.,
Chemical Enhancers and Transdermal NitroDur from Schering-Plough, Nicotrol from
Patch Technologies Pfizer), the adhesive contains the drug and provides
adhesion of the patch to the skin. The patch
Two main types of transdermal patches are commonly utilizes the skin as the rate-controlling membrane.
used: drug reservoir patch and drug-in-adhesive patch. The advantages of a drug-in-matrix system are larger
Transdermal patches of early generations are mainly diffusional surface area for drug delivery than a drug
 3846 Transdermal Delivery: Technologies

reservoir system, thinner patch system, simpler con- and mechanistic approaches such as examining the quan-
struction, and good skin conformability. However, titative structure enhancement relationships of chemical
not all drugs and formulations are compatible with penetration enhancers have been the topics of interest
the adhesives. Because an optimal patch system is recently.[15,16]
usually small, it requires high drug concentrations in
the patch. The compatibility issue with the adhesives
Carriers and Vehicles
limits the concentration and the quantity of drug in
the patch. Possible incompatibility between chemical
Vehicles and carriers such as liposomes, micelles,
enhancers and adhesive also creates challenges and
synthetic nanoparticles, and cyclodextrin can be used
problems to incorporate chemical permeation enhan-
in transdermal patches and topical formulations to
cers in this system. Examples of transdermal patch sys-
increase drug solubility and stability. Such increase in
tems available in the market or under development are
solubility leads to higher drug loading capacity, and
3M Lattitude transdermal system (3M Drug Delivery)
hence, smaller transdermal patches. In addition to
such as Minitran and Climara, D-Trans system (Alza
solubilization, carriers can also improve drug stability
Corp) such as Estraderm and Testoderm, dot matrix
and have been suggested to enhance transdermal
system (Noven Pharmaceuticals) such as Vivelle Dot
absorption.[17] However, the mechanisms of carrier-
and Estradot, matrix type systems for Nicotinell
enhanced transdermal transport remain controversial.
(Novartis), Nitrodisc (Roberts Pharmaceuticals), and
It is generally believed that carriers such as liposomes,
hydrogel patch system such as Lidoderm (Endo Phar-
micelles, and cyclodextrin, due to their sizes, do not
maceuticals). New patch designs and transdermal for-
readily penetrate the stratum corneum[18] but smaller
mulations mainly aim to provide better transdermal
molecules such as phospholipids and surfactants (the
flux enhancement, less irritation and adverse skin reac-
components forming liposomes and micelles) can pen-
tions, convenience, and comfort. In addition to the
etrate the stratum corneum and alter the phase beha-
patches, transdermal gel systems such as AndroGel
vior of the stratum corneum intercellular lipids.[19–21]
and EstroGel for testosterone and estradiol transder-
Transfersome (IDEA AG) is a carrier system currently
mal delivery (Solvay Pharmaceuticals), respectively,
under development for transdermal applications. It has
Testim (Auxilium Pharmaceuticals), and LibiGel
been suggested that these ‘‘deformable’’ vesicles/
Trace–Ultrasonic

(Antares Pharma) are also available.

carriers can penetrate the skin under certain con-
ditions[22] and enhance transdermal absorption.[23] It
is also claimed that lipid vesicles can penetrate the
Chemical Penetration Enhancers
skin if they are sufficiently deformable, but direct evi-
dence for the penetration of intact unaltered carriers/
There are many varieties of chemical penetration
vesicles across the stratum corneum remains to be seen.
enhancers for transdermal transport. It is not practical
Carriers can also reduce the effects of the unstirred
to discuss all these enhancers here. The readers should
layer in the drug chamber of a transdermal system,
refer to other references on this topic. Generally, it is
alter the stratum corneum barrier via lipid extraction,
difficult to clearly define the effects of the excipients
protect the drug against skin metabolism in the drug
and categorize these excipients in a transdermal patch.
compartment, and/or effectively deliver chemical
For example, the Androderm patch (Watson Pharma-
enhancers to the stratum corneum.[24,25] Recent studies
ceuticals) contains the drug testosterone in alcohol,
have also shown enhanced transdermal drug delivery
glycerin, glycerol monooleate, and methyl laurate in
with the conjugation of the drug to specific peptides.[26]
an acrylic acid copolymer. All these excipients have
both solubilizing and penetration enhancing effects.
Among them, the long alkyl chain compounds glycerol Physical Enhancer
monooleate and methyl laurate are more effective
enhancers than alcohol.[11,12] In general, chemical Iontophoresis
permeation enhancers increase drug absorption by
increasing the permeability coefficient of skin via Transdermal iontophoresis is the enhanced transport
mechanisms that mainly increase KL and DL for lipo- of drug across skin by an externally applied electric
philic permeants and eP and DP for polar permeants field. An iontophoresis device has at least two electro-
[Eqs. (4) and (5)]. Although there has been progress des, each positioned so as to be in electrical contact
in understanding the mechanisms of permeation with some area of the body surface. Generally, one
enhancers, screening remains the primary method to electrode is the donor electrode that contains the drug
select appropriate enhancers in the development of to be delivered. The other electrode, referred to as the
transdermal formulations.[13] In this approach, high counter or return electrode, serves to close the electri-
through-put screening is desirable.[14] More systematic cal circuit through the body and is placed next to the
Transdermal Delivery: Technologies 3847

first electrode. An electrical circuit is formed by the membrane counterion, the properties of the drug
connection of these electrodes to a source of electrical (e.g., surface active drug) being delivered across the
energy such as a battery and circuitry capable of membrane, and the properties of the excipients (e.g.,
controlling the amount of current passing through the surfactants) in the formulation.[32–34] Electroporation
device and body surface. Table 2 provides a list of trans- by definition is the permeabilization of a mem-
dermal iontophoresis companies which manufacture and brane under an electric field. Under this definition,
develop iontophoresis devices, and have an iontophor- electroporation can occur in conventional transdermal
esis product in the market or in late stage development. iontophoresis[35] as evidenced by the decrease of skin
A more complete list of animal and human clinical electrical resistance and the increase in the intrinsic
studies can be found in the literature.[27,28] skin permeability during iontophoresis [eP and DP in
The mechanisms of transdermal iontophoresis are Eq. (6)]. It has been observed that electric fields of
electrophoresis, electroosmosis, and skin alteration. greater than 0.5 V alter the structure of the stratum
Electrophoresis is the facilitation of the movement of corneum and its intrinsic permeability during ionto-
ionic species by the applied electric field[29] following phoresis.[36] Such alteration can be characterized as
the principle of Nernst-Planck theory [the second term reversible or irreversible and is related to the magni-
in Eq. (6)]. The electric field causes the ions to move. tude and duration of the applied electric field (i.e.,
Positively charged ions move from the positive elec- the electric current).
trode (anode) and negatively charged ions move from The efficiency of iontophoretic transport is usually
the negative electrode (cathode). When the therapeutic assessed by the transference number (ti) when ionto-
compound is positively charged, the anode acts as the phoresis is operated at constant current. The transference
active electrode and the cathode serves as the counter number defines the fraction of the current carried by an
electrode. Conversely, when the therapeutic ion is ionic species of interest (i.e., the permeant or drug ion).
negatively charged, the cathode is the active electrode It is the ratio of the current carried by the permeant (Ii)
and the anode is the counter electrode. Electroosmosis to the total current carried by all ionic species (Itotal) in
is the movement of solvent along a charged surface the system:[27,28,37,38]
under the influence of an electric field [the third term
in Eq. (6)] arising from the momentum of the excess Ii
ti ¼ ð7Þ

Trace–Ultrasonic
counterion in the solvent (i.e., higher concentration Itotal
of the counterion than the coion due to the charged
surface[30,31]). It enhances the movement of both neu- and
tral and positively charged species across skin under jzi jJi
normal physiological conditions because the stratum ti ¼ P   ð8Þ
zj J j
corneum is negatively charged under these conditions. j
Thus, the effect of electroosmosis is pH dependent.
Because transdermal electroosmosis is an electrokinetic where Ji is the flux of species i (the permeant), Jj is the flux
phenomena of a charged surface, the effect of electro- of ionic species j in the system, Ii is the current carried by
osmosis is also affected by the ionic strength of the ionic species i, Itotal is the total current, zi is the charge of
solution in the skin membrane, the properties of the the ionic species i, and zj is the charge of ionic species j.

Table 2 Examples of companies in iontophoresis

Company Technology/system Drug product Address
Alza E-trans Fentanyl ww.alza.com
a
Birch Point Medical WEDD, IontoPatch — www.birchpoint.net
Cygnus/Animas GlucoWatch Biographer Glucose monitoring www.glucowatch.com
Empi Dupel, Action Patch — www.empi.com
Life Tech Iontophor, Microphor — www.life-tech.com
Iomed Phoresor Lidocaine www.iomed.com
General Medical Co Lectro, Drionic — www.drionic.com
Transport Pharmaceuticals — Acyclovir www.transportpharma.com
Vyteris LidoSite Lidocaine www.vyteris.com
a
The WEDD transdermal system does not apply a high enough electrical potential to generate the electric current commonly observed in trans-
dermal iontophoresis. Although the electric field may provide some flux enhancement, flux enhancement due to the electric current in this system
is limited. The system therefore may not be viewed as a transdermal iontophoresis system.
 3848 Transdermal Delivery: Technologies

The ionic species j represents both the ions migrating a ready-to-use system (e.g., IONSYSTM E-TRANSÕ
into the receiver from the donor including ionic species system, Alza Corp., CA). In the development of a
i and the oppositely charged counter-ions migrating ‘‘wet’’ system, the compatibility and stability of the
into the donor from the receiver. In dominant electro- drug, formulation, drug compartment matrix, and
transport (negligible passive diffusion contribution) and electrode system must be considered. For example,
combining the electroosmosis and electrophoresis terms the drug solution can interact with the electrode or
in Eq. (6), Eq. (9) is commonly seen in the iontophoresis metallic electrodes can act as catalyst in drug degra-
literature: dation during storage. The electronic and battery unit
can be disposable after a single use (e.g., IONSYS
jzi jm Ci E-TRANS system) or reusable (e.g., Phoresor system).
ti ¼ P  i ð9Þ
zj mj C j It is more cost effective to have a separate reusable
j
electronic and battery unit (from the drug chamber)
where mi and mj are the mobilities of the ionic species i but a single use disposable unit (electronics combined
and j, Ci and Cj are the concentration of the permeant with the drug chamber) is more user-friendly and
i and j in the membrane, respectively. ‘‘fool-proof.’’ Although miniaturization, electronics,
The flux of the permeant (Ji) is related to the and battery technologies have advanced tremendously
current by: in the past decade and should not pose a problem in the
design of an iontophoresis device, chemistry-related
ti Itotal issues such as stability and compatibility continue to
Ji ¼ ð10Þ
AD F jzi j be important and can be challenging in transdermal
iontophoresis development.
where F is the Faraday constant. From Eqs. (7)–(10), it
can be seen that the flux of the permeant is pro- Iontophoresis electrode technology
portional to the electric current and the transference
number. The transference number is a function of the To optimize transdermal iontophoretic transport, the
fluxes of all ion species transporting across the mem- concentration of drug in the donor chamber should
brane, which is related to the concentration of the be maximized and the concentration of the co-ions
Trace–Ultrasonic

ion species in the drug chamber and in the body (background electrolyte) in the chamber should be
(endogenous ions) at the iontophoresis application site. minimized [see Eqs. (7) and (9)]. Significant changes
in pH and changes in solution composition in the drug
Iontophoresis devices chamber can also occur due to ion transport and the
formation of electrochemical products at the electrode.
Iontophoretic transport can be influenced by different Inert or non-sacrificial (catalytic) electrodes such as
factors. Not all drugs can be effectively delivered with Pt and carbon can induce significant pH changes:
iontophoresis depending on the molecular size, structure, hydrogen ion is generated at the anode and hydroxide
and charge. Issues related to device development include is generated at the cathode. An estimation using the
concentration of the drug in the drug compartment, Faraday’s law shows that an iontophoresis application
background electrolytes in the compartment, pH and of 0.2 mA for one hour will generate around 7 mM
ionic strength of the compartment, barrier properties of of hydroxide ion in a 1-mL electrode chamber at the
the skin, electric current density, and total electric cur- cathode. This corresponds to a change of pH from
rent. Other issues that need to be considered include 7 to 12 without a buffer. Because the use of inert
the electrode type, battery life, drug compartment matrix, electrode is not desirable, commercial products usually
stability of the matrix and electrode system in storage, employ a sacrificial electrode system such as Ag/AgCl
adhesive, electrochemical reaction at the electrode during electrodes.[40,41] The Ag/AgCl electrode system is not
iontophoresis, and patient factors. For example, if the without limitations. Ag electrodes require chloride or
electrode configuration is not properly designed, skin similar ions to sustain the electrochemical reaction at
electrochemical burn could be observed after sustained the anode, and Ag/AgCl electrodes produce chloride
DC iontophoresis due to the accumulation of hydrogen ion at the cathode. If the chloride ion at the anode is
ion at the anode and hydroxide ion at the cathode.[39] not sufficient to sustain the electrode system, Ag ion
In the design of a transdermal iontophoresis system, will be generated, which can cause Ag stain (Ag burn)
several factors must be considered. First, the system on the skin. Thus, in iontophoretic delivery of cationic
can be a ‘‘wet’’ or ‘‘dry’’ system. In a ‘‘dry’’ system, the drugs, it is preferred to use the chloride salt form of
drug solution is separated from the electrode and the the drugs[42] to sustain the electrochemical reactions
applicator shell in storage. The drug solution is added and limit the amount of competing ions in the drug
to the applicator before use (e.g., Numby StuffÕ chamber. Hydrogen ion, hydroxide ion, chloride ion,
PhoresorÕ system, Iomed Inc., UT). A ‘‘wet’’ system is or Ag ion generated at the electrode will also compete
Transdermal Delivery: Technologies 3849

with the transference of the drug ion and reduce the generally used in systematic and mechanistic study of
efficiency of iontophoretic delivery. Several technolo- iontophoretic transport. Thus, constant voltage DC
gies have been proposed to overcome these problems. iontophoresis is not commonly employed in practice
For example, permselective membranes or materials but is mainly found in academic setting in the literature.
can be used to isolate the drug chamber from the elec- Alternating current (AC) is also employed in ionto-
trode and prevent the migration of the electrochemical phoretic transport. For example, the GlucoWatchÕ
products at the electrode into the drug chamber.[43] An G2TM Biographer for glucose monitoring (Cygnus
ion-exchange resin (or material) loaded with chloride Inc., CA) utilizes an iontophoresis protocol that alters
or other Ag precipitating materials can be incorpor- the polarity of a DC in each ten-minute cycle. Lectro
ated in the electrode chamber to either supply sufficient PatchÕ (General Medical Co., CA) comprising a pair
amounts of chloride ion for the electrochemical of carbon or silicon electrodes also operates under
reaction at the anode or remove the Ag ion generated AC at a frequency of between 0.0027 and 10 Hz for
at the electrode, respectively.[44,45] The ion-exchange drug delivery. It is suggested that AC can eliminate
resin or material can also be coated on the conductive potential electrochemical burn and reduce skin
electrode.[46] Other methods have been proposed to irritation and sensation that would otherwise occur
utilize specific membranes and materials to precipitate during long iontophoresis application or under the
or bind the chloride ion at the cathode.[44,47] Another circumstances when an inappropriate electrode design
use of an ion-exchange resin or membrane in the drug is used.[39,52,53] Other advantages of AC are its ability
chamber is to increase the drug loading capacity to to reduce transdermal flux variability of neutral
enhance iontophoresis efficiency and reduce the size permeants (electroosmosis) and to maintain the skin
of the device.[48–50] Ion-selective membrane can also electric resistance constant. Significant inter-subject
be interposed between the drug reservoir and the skin and intra-subject variability and flux drift (the increase
surface that only allows passage of the drug ion and in permeant flux with time) are observed in transder-
ions having the same charge as the drug but prevents mal iontophoretic transport of neutral polar per-
the passage of the ions of the opposite charge (to the meants,[54–58] and iontophoresis operating under the
drug) that migrate from the body into the drug AC principles has been shown to reduce such varia-
reservoir.[51] The exclusion of the endogenous ions bility. It is also found that AC enhances transdermal

Trace–Ultrasonic
carrying the electric current from the body into the flux by the mechanism of electroporation similar to
reservoir can enhance the transference of the drug that that occurs during DC. Transport enhancement
and drug delivery. of AC over that of electroporation is related to electro-
phoresis and electroosmosis due to the asymmetric
Direct current and alternating current boundary conditions in the drug chamber (donor)
and the body (receiver) across the membrane.[59,60]
Iontophoretic devices generally employ a constant
direct current (DC) approach. Examples of constant Electroporation
current iontophoresis device are Numby Stuff Phoresor
system (Iomed, Inc., UT) and LidoSiteTM Topical Transdermal electroporation is a method to induce
system (Vyteris, Inc., NJ) for local lidocaine delivery transient small pores in the skin to enhance transder-
and IONSYS E-TRANS system for systemic fentanyl mal drug delivery with the application of high electric
delivery (Alza Corp., CA). The transference number field pulses. Although pores are also induced during
[as defined in Eq. (7)] is generally assumed to be con- conventional iontophoresis,[35,36] transdermal electro-
stant in normal iontophoresis operation. As a result, poration is commonly defined as the application of
it is believed that a constant current would lead to con- short (millisecond) high voltage pulses 50 V.[61]
stant permeant flux, and permeants could theoretically Similar to iontophoresis, the flux enhancing mechan-
be delivered or extracted at a constant, predictable, isms of electroporation are electropermeabilization
and programmable rate that is only dependent upon (membrane alteration, pore induction), electrophoresis
the applied current. For a small ionic molecule, (direct-field effect), and electroosmosis (electric field
although inter- and intra-subject variation of the trans- induced convection) with electropermeabilization
ference number (i.e., permeant flux) has been observed being the major flux enhancing mechanism [i.e.,
during iontophoresis, such variability is usually within increasing ep and Dp in Eq. (6)]. Electropermeabiliza-
acceptable range with an adequate electrode design. tion is the spontaneous formation of metastable pores
Another DC approach is to maintain constant voltage from the energy of the electric field. The minimum
(instead of constant current) during iontophoresis. voltage or electric field required to induce pore forma-
During constant voltage iontophoresis, the skin intrinsic tion is around 0.2–1.0 V per lipid bilayer, depending
permeability continues to change, making the control of on the bilayer composition. The rate of pore formation
the iontophoretic flux complicated. This approach is is energy and field dependent and the rate of pore
 3850 Transdermal Delivery: Technologies

destruction (membrane recovery) is usually assumed to in 5–15 minutes of application.[64] Review articles of
be independent of the field. During electroporation, microneedles in transdermal transport are available
new electric current pathways are created in the stra- in the literature.[65]
tum corneum, which can be reversible or irreversible.
At greater than 30 V across the stratum corneum,
electroporation is suggested to occur at the lipid- Poration with heat, laser, and RF
corneocyte matrix, whereas at voltage less than 30 V,
electroporation is believed to occur at the appendageal The PassPort system from Altea Therapeutics uses a
ducts, e.g., cells lining the appendages.[35] Details of thermal process to create an array of macropores
the mechanisms and applicability of electroporation across the stratum corneum. The device consists of a
in transdermal transport can be found in the litera- planar array system of small metallic filaments that
ture.[62] Potential problems of electroporation in trans- rapidly converts electrical energy to thermal energy.
dermal delivery are similar to those of iontophoresis The heat ablates the stratum corneum for a few
with more severe problems related to sensation, tissue milliseconds at a time and porates only the stratum
damage, and muscle reflex under the high electric field corneum. After skin poration with this process, drug
pulses. To reduce electric current traveling across the absorption occurs from a drug reservoir on the porated
muscle, microarray electrodes are being developed to site. Hydromorphone and insulin delivery with this
avoid deep electric current penetration during method is currently under human clinical testing.
electroporation.[63] Laser such as ArF (argon–fluoride) at short ultra-
violet wavelength and erbium : YAG at wavelength
strongly absorbed by tissue water can be used to create
Poration and Physical Abrasion pores in the stratum corneum. The main obstacles of
this approach are the hardware cost of laser and the
Owing to transport hindrance related to the steep- precision of the poration process. SpectRx Inc is cur-
molecular size dependence of transdermal transport, rently developing a continuous glucose monitor for
several technologies have been studied to physically people with diabetes after precise abrasion with laser.
remove the stratum corneum [i.e., increasing eP and DP In this application, the interstitial fluid is collected
through an array of microscopic holes, or micropores,
Trace–Ultrasonic

in Eq. (6)]. Micron-scale needle (microneedle), laser,

and radio frequencies (RF) current are among the created with a laser in the stratum corneum. Norwood
technologies under development in this area. The main Abbey has been developing a handheld device that dis-
goal of these technologies is to create small micropores rupts the stratum corneum with a burst of low-level
across the stratum corneum without penetration into laser light. With the low level laser, it is suggested that
the dermis and the nerve ending in the skin. Thus, these the device only alters the stratum corneum structure
methods are suggested to be painless, minimally invas- and does not punctuate the stratum corneum.
ive, and within the limits of toleration. RF wave, used in electro-surgery, is also applied to
create macropores in the stratum corneum. These
Microneedle macro-passages allow effective delivery of drugs and
extraction of endogenous compounds from the body
With the advances in microfabrication technology in surface. TransPharma Medical is developing a hand-
microelectronics industry, arrays of micron-sized nee- held electronic unit and a disposable microelectrode
dles can now be manufactured and produced on a array system utilizing this technology for transdermal
massive scale. The proposed uses of microneedle patch transport. The device has a feedback system that con-
include vaccination and the delivery of macromole- trols the duration of the poration process by monitor-
cules such as oligonucleotides and peptides. Solid ing the electric current across the system. The poration
needle arrays for penetrating the stratum corneum to process is automatically terminated when macropores
create macropores that increase transdermal transport reach the viable epidermis.
or drug-coated needle arrays for carrying the drug into
the skin have been investigated. Hollow microneedles Heat
that remain on the skin surface during transdermal
application are also under development. An example The mechanisms of heat-assisted transdermal delivery
is the Alza Macroflux patch. Macroflux is a drug- include an increase in drug diffusion, increase in skin
coated titanium microneedle array. The tips of micro- permeability, alteration of skin structure, increase in
needle arrays are covered with a solid coating of the body circulation, and increase in the release rate of
drug crystal. When applied to the skin, the drug-coated drug from local skin tissue into systemic circulation.
needles penetrate the skin and deliver the drug from Heat can also increase drug solubility. The advantage
the drug-coated needle surface into the epidermal layer of heat control is that it does not involve chemical
Transdermal Delivery: Technologies 3851

and physical enhancers that may cause skin irritation testosterone for hormone replacement therapy, fenta-
and other adverse skin effects. It can modify drug nyl for pain management, nicotine for smoking cess-
absorption on an on-demand basis such as to treat ation, nitroglycerin for angina pectoris,
breakthrough symptoms of pain and anxiety and be norelgestromin and ethinyl estradiol as contraceptive,
removed to slow down drug absorption. By increasing and scopolamine for motion sickness. These products
the rate of drug absorption, shorter onset time of drug account for a market of more than $2 billion in the
action is also expected. Because no chemical permeation United States. Among these therapeutic areas, pain
enhancer is used, component compatibility issues are also management (e.g., fentanyl for chronic pain and lido-
avoided. If chemical enhancers are used, heat can shorten caine for postherpetic neuralgia) and hormone replace-
the onset time of the enhancer effects. An example of ment (e.g., estradiol and testosterone) are the largest
heat-assisted transdermal systems is controlled heat- and fastest growing areas. Other attractive areas
assisted drug delivery (CHADD) by Zars. The main include the delivery of macromolecules such as oligo-
component of the CHADD system is a patch-like chemi- nucleotides and peptides and vaccination targeting
cal heating layer of heat-generating chemicals that the immune systems such as Langerhans cells in the
enhance the penetration of drug. The temperature and viable epidermis. Despite the enormous interest in
duration of the heating layer is predetermined ranging these areas, small molecules remain the mainstream
from 5 minutes to 24 hours to maintain the temperature candidates of transdermal delivery due to the barrier
of the skin and the underlying tissues to within a set tem- properties of the stratum corneum. Another development
perature range (from 31 C to 41 C) for that time period. in the transdermal industry that is worth mentioning is
transdermal analyte monitoring and the anticipated
development of automatic feedback systems such as the
Ultrasound
combination of transdermal blood glucose monitoring
and insulin delivery (artificial pancreas) in the manage-
Sonophoresis (ultrasound) enhances transdermal trans-
ment of diabetes. Along this development, a device
port through the mechanisms of pore induction
of transdermal iontophoretic glucose extraction and
(membrane alteration due to cavitation), convection,
monitoring was introduced in mid-2002 (GlucoWatch,
and stratum corneum lipid alteration. Potential pro-
Cygnus).
blems of the sonophoresis approach include the signifi-

Trace–Ultrasonic
New technologies are usually first tested with
cant increase in temperature at the site of application
approved small molecules in well-established thera-
during sonophoresis, the requirement of the coadminis-
peutic areas for the proof of concept. For example,
tration of chemical enhancers such as sodium dodecyl
the first FDA approved transdermal sonophoresis
sulfate (SDS) for effective transdermal transport in
product (SonoPrep by Sontra Medical) in Aug 2004
vivo, and unpredictable flux due to inter-subject and
topically delivers lidocaine to anesthetize skin before
intra-subject variability of the energy required to
patients undergo painful skin procedures such as nee-
achieve the same stratum corneum permeability state.
dle pricks and the insertion of catheters. Similarly,
Detailed information regarding the mechanisms and
transdermal iontophoretic delivery of lidocaine (with
the interplay of frequency and energy of sonophoresis
epinephrine) to provide local skin analgesia is the only
in transdermal transport can be found elsewhere.[66]
FDA approved indication for iontophoresis at this
The SonoPrep system by Sontra Medical operates
time (Iontocaine by Iomed and LidoSite by Vyteris
under the principle of sonophoresis. The company has
in Summer 1996 and Spring 2004, respectively). Lido-
recently completed several clinical studies with the
caine is also the drug of choice in the first NDA fillings
system. The SonoPrep system uses a short burst (about
of heat-assisted transdermal delivery (S-Caine Patch
15 seconds) of ultrasound (55 kHz) to alter the stratum
and S-Caine peel by Zars, NDA filed in Mar 2003)
corneum in a pretreatment process to enhance drug
when a systemic fentanyl CHADD system is still in
delivery. The first application of the SonoPrep system
clinical trials. Another iontophoresis product, which
is to locally deliver lidocaine that permits faster onset
received an approval letter from FDA in Jul 2004, is
time of pain relief prior to painful skin procedure.
the E-TRANS fentanyl HCl patient-controlled system
According to the company, the system provides skin
for acute pain management (IONSYS by Alza). The
anesthesia in approximately five minutes after SonoPrep
device is small and is applied to the patient’s upper
pretreatment vs. 60 minutes without the pretreatment.
outer arm or upper chest. It has a button for the patients
to activate the system and deliver fentanyl for break-
Therapeutics through pain. The system has been shown to provide
the same effect as patient-controlled intravenous fentanyl
Currently approved therapies of systemic drug delivery delivery but is needle-free and more convenient to
via the transdermal route include aprostadil for erectile patients. The device is currently targeted for in-hospital
dysfunction, clonidine for hypertension, estradiol and use only, according to the NDA filing.
 3852 Transdermal Delivery: Technologies

CONCLUSIONS 2. Flynn, G.L. Mechanism of percutaneous absorption from

physicochemical evidence. In Percutaneous Absorption;
Bronaugh, R.L., Maibach, H.I., Eds.; Marcel Dekker:
Technology in medicine is defined as the application New York, 1985; 17–52.
and evolution of basic science to clinical practice. It is 3. Ackermann, C.; Flynn, G.L.; Smith, W.M. Ether-water
important to understand that the advance in trans- partitioning and permeability through nude mouse skin in
vitro. II. Hydrocortisone 21-n-alkyl-esters, alkanols and
dermal technologies depends largely on our understand- hydrophilic compounds. Int. J. Pharm. 1987, 36, 67–71.
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transport and of the methods to enhance it. After trans- Y. Effect of vehicle on the skin permeability of drugs:
polyethylene glycol 400-water and ethanol-water binary
dermal drug delivery was re-introduced into modern solvents. J Controlled Release 1993, 23, 247–260.
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1990s. In recent years, we have observed progress in group chain length and polar head group on chemical
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identification of more therapeutic areas suitable for the S.K. Mechanistic studies of the 1-alkyl-2-pyrrolidones
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can be long and difficult. Nevertheless, drug delivery diffusion of polar molecules through and effective pore
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mal membrane. Pharm. Res. 1994, 11, 1306–1314.
delivery technologies and more new transdermal pro- 11. Patel, D.C.; Chang, Y. Penetration Enhancement with Binary
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ducts are expected in the near future. This is a result System of Oleic Acid, Oleins, and Oleyl Alcohol with Lower
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12. Walters, K.A.; Hadgraft, J. Pharmaceutical Skin Pen-
etration Enhancement; Marcel Dekker: New York, 1993.
13. Karande, P.; Jain, A.; Ergun, K.; Kispersky, V.; Mitragotri,
S. Design principles of chemical penetration enhancers
ACKNOWLEDGMENTS for transdermal drug delivery. Proc. Natl. Acad. Sci. USA
2005, 102, 4688–4693.
The author thanks Dr. William I. Higuchi, David 14. Karande, P.; Mitragotri, S. High throughput screening of
transdermal formulations. Pharm. Res. 2002, 19, 655–660.
J. Miller, and Ned Weinshenker for their helpful 15. Ghafourian, T.; Zandasrar, P.; Hamishekar, H.; Nokhodchi,
discussions. A. The effect of penetration enhancers on drug delivery
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16. He, N.; Warner, K.S.; Chantasart, D.; Shaker, D.S.;
Higuchi, W.I.; Li, S.K. Mechanistic study of chemical skin
ARTICLES OF FURTHER INTEREST permeation enhancers with different polar and lipophilic
functional groups. J. Pharm. Sci. 2004, 93, 1415–1430.
17. Manosroi, A.; Kongkaneramit, L.; Manosroi, J. Stability
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Drug Delivery: Topical and Transdermal Hashem, F.M.; Higuchi, W.I. Mechanistic studies of the
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Polymers in Transdermal Delivery Systems, p. 2925. 19. Bouwstra, J.A.; Honeywell-Nguyen, P.L. Skin structure and
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20. Kirjavainen, M.; Monkkonen, J.; Saukkosaari, M.;
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iontophoresis. Adv. Drug Del. Rev. 1992, 9, 201–237. 55. Tamada, J. Method of Sampling Substances Using Alter-
31. Peck, K.D.; Srinivasan, V.; Li, S.K.; Higuchi, W.I.; nating Polarity of Iontophoretic Current. US Patent
Ghanem, A.-H. Quantitative description of the effect of 6,023,629,February 8, 2000.
molecular size upon electroosmotic flux enhancement 56. Zhu, H.; Li, S.K.; Peck, K.D.; Miller, D.J.; Higuchi, W.I.

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during iontophoresis for a synthetic membrane and human Improvement on conventional constant current DC ionto-
epidermal membrane. J. Pharm. Sci. 1996, 85, 781–788. phoresis: a study using constant conductance AC ionto-
32. Peck, K.D.; Hsu, J.; Li, S.K.; Higuchi, W.I.; Ghanem, A-H. phoresis. J. Control Release. 2002, 82, 249–261.
Flux enhancement effects of ionic surfactants upon passive 57. Li, S.K.; Higuchi, W.I.; Kochambilli, R.P.; Zhu, H. Mech-
and electroosmotic transdermal transport. J. Pharm. Sci. anistic studies of flux variability of neutral and ionic
1998, 87, 1161–1169. permeants during constant current DC iontophoresis
33. Hirvonen, J.; Guy, R.H. Iontophoretic delivery across the with human epidermal membrane. Int. J. Pharm. 2004,
skin: electroosmosis and its modulation by drug substances. 273, 9–22.
Pharm. Res. 1997, 14, 1258–1263. 58. Li, S.K.; Higuchi, W.I.; Zhu, H.; Kern, S.E.; Miller, D.J.;
34. Hirvonen, J.; Guy, R.H. Transdermal iontophoresis: modu- Hastings, M.S. In vitro and in vivo comparisons of
lation of electroosmosis by polypeptide. J. Control Release constant resistance AC iontophoresis and DC ionto-
1998, 50, 283–289. phoresis. J. Control Release 2003, 91, 327–343.
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Galichenko, S.V.; Weaver, J.C.; Potts, R.O. Electrical Quantitative study of electrophoretic and electroosmotic
properties of skin at moderate voltages: contribution of enhancement during alternating current iontophoresis
appendageal macropores. Biophys. J. 1998, 74, 843–856. across synthetic membranes. J. Pharm. Sci. 2004, 93,
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effects of applied voltage and duration on human epidermal 60. Mollee, T.R.; Anissimov, Y.G.; Roberts, M.S. Periodic
membrane alteration/recovery and the resultant effects electric field enhanced transport through membranes.
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Enhancers; Smith, E.W., Maibach, H.I., Eds.; CRC Press: electroporation: comparisons and contrasts. Int. J. Pharm.
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Drug Deliv. Rev. 1992, 9, 137–176. transdermal and topical delivery. Adv. Drug Deliv. Rev.
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Drug Delivery. US Patent 5,415,628, May 16, 1995. desmopressin using a coated microneedle array patch sys-
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 Ultrasonic Nebulizers
Kevin M.G. Taylor
School of Pharmacy, University of London, London, U.K.

Orla McCallion
Vandsons Research, London, U.K.

INTRODUCTION to produce bronchodilator responses comparable with

those of air-jet nebulizers and pMDIs.[8]
Ultrasonic nebulizers use ultrasonic energy to convert
liquid, usually an aqueous solution, into an aerosol
for inhalation. They are used to deliver b2-agonists,
MECHANISM OF AEROSOL GENERATION
corticosteroids, antiallergics, anticholinergics, and
antiviral and mucolytic agents to the respiratory
The energy required to atomize a liquid is produced by
tract.[1] Recent innovations have increased the popu-
a piezoelectric crystal transducer, usually a synthetic
larity of nebulizers, both ultrasonic and air-jet, and
ceramic material, vibrating at a high frequency
new devices with improved portability, compared with
(1–3 MHz). When an alternating electric current is
traditional models, are capable of generating aerosols
applied, the crystal shrinks and expands and the resul-
with high respirable fractions (deep lung delivery) with
tant vibrations are transmitted to the nebulized fluid,
a high drug output. Nebulized drugs may be inhaled
either directly or via a coupling liquid, usually water.
during normal tidal breathing through a mouthpiece
A fountain of liquid is produce at the liquid surface,
or face mask, permitting their use for patients, such
with large droplets being emitted from its apex and a
Trace–Ultrasonic

as the hospitalized, the elderly, children, and patients

‘‘fog’’ of small droplets being produced from the lower
with arthritis, who experience difficulties with other
part.
devices. Nebulizers represent ideal delivery systems
Fig. 1 illustrates the two mechanisms proposed for
for drugs that cannot be conveniently formulated into
the processes of liquid disintegration and aerosol gen-
pressurized metered-dose inhalers (pMDIs) or dry
eration within ultrasonic nebulizers.[9] The capillary-
powder inhalers (DPIs) or when the therapeutic dose
wave theory relates to the production of capillary
is too large for delivery with these systems.
waves in the bulk liquid. These waves constructively
Ultrasonic nebulizers were first developed in the
interfere to form peaks and a central geyser. When
1960s and were initially used for air humidification in
the amplitude of the applied energy is sufficiently high,
respiratory care units.[2] Generally, ultrasonic nebulizers
the crests of the capillary waves break off, and droplets
have higher mass outputs than do air-jet nebulizers, in
are formed. The rate of generation of capillary waves is
which compressed gas is used as a means of generating
dependent on both the physicochemical properties of
the aerosol. However, this is achieved at the expense
the nebulized fluid and the intensity of the ultrasonic
of a large aerosol droplet size.[3–7] Consequently, the
vibration. Mercer[3] used Eq. (1) to calculate the
majority of ultrasonically generated aerosols have a
threshold amplitude for the generation of capillary
mean droplet size considered unsuitable for efficient
waves:
targeting of a drug to the alveolar region of the lung.
However, the size distribution of droplets within aero-
v
sols produced by ultrasonic nebulization is generally A ¼ 4 ð1Þ
less than that from air-jet nebulizers. Nevertheless, fl
the generated aerosols are polydisperse in nature, and
as with air-jet nebulizers, they require baffles to remove where A is the threshold amplitude, v is the kinematic
the larger droplets from the emitted aerosol. Because viscosity of the liquid, f is the acoustic frequency, and
ultrasonic nebulizers have a high mass output, with l the capillary wavelength. When the amplitude
increased droplet concentration, which is independent exceeds the threshold value by a factor of approxi-
of the airflow, the duration of nebulization is shorter mately four, droplets are formed. Lang[10] noted that
than with air-jet nebulizers. This, together with their the mean droplet size generated from thin liquid layers
quieter mode of operation, may make them particularly was proportional to thecapillary wavelength on the
attractive to patients. Ultrasonic nebulizers are reported liquid surface. Using an experimentally determined
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-100000444
3854 Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved.
Ultrasonic Nebulizers 3855

A B NEBULIZER DESIGN

Ultrasonic nebulizers exist in a number of basic designs

that differ in the configuration of the piezoelectric crys-
tal transducer, nebulizer chamber, baffles, and auxil-
iary airflow systems (Fig.2).[9] Once the aerosol cloud
is generated from the nebulizer fluid, it is transferred
from the chamber and made available to the patient.
Ultrasonic nebulizers produce a large number of drop-
lets per unit volume, which tend to aggregate and settle
in the absence of air circulating through the device.
Larger droplets impact on the baffles or internal surfaces
to return to the reservoir surface for recirculation under
the influence of gravity. Smaller droplets leave the device
aided by an internal fan (e.g., Medix ElectronicÕ,
EasimistÕ) or by entrainment into the inspiratory flow
Fig. 1 Proposed ultrasonic nebulizer aerosolization mechan- of the patient (e.g., DeVilbiss PulmosonicÕ). Air velocity
isms. (A) Cavitation bubble formation at low frequency; (B) over the reservoir surface may be modified by fan speed
capillary-wave formation at high frequency. (From Ref.[9].) (and flow constrictors), thereby influencing both droplet
size and aerosol output rate. For instance, changing the
factor of 0.34, the droplet diameter D is given by fan speed setting of the Sonix 2000Õ (Medix) ultrasonic
Eq. (2); nebulizer can vary flow output between 1 ml per min and
1 ml per 6 min. The important design features influencing

D ¼ 0:34l ð2Þ

where D is the number median diameter and l is the

Trace–Ultrasonic
A
capillary wavelength. Lobdell[11] concurred with these
findings and calculated a theoretical value of 0.36 for
the proportionality constant. The capillary wavelength
can be calculated from Kelvin’s equation, as in Eq. (3):

l ¼ ð8py=rf 2 Þ1=3 ð3Þ

B

where g is the surface tension, r is the density, and f the

acoustic frequency. When y is in mN/m (dyne/cm), r
C
is in g/cm3, and f is in megacycles/s, then D is given
in micrometers. Good correlation exists between D
calculated and experimentally derived values.[12]
The second mechanism proposed for aerosol gener-
ation is based on the piezoelectric crystal operating at
low frequency and imparting vibrations to the bulk
liquid. This results in the formation of cavitation bub-
bles, which move to the air–liquid interface.[13] The
internal pressure within the bubbles equilibrates with
that of the atmosphere, causing their implosion. When
this occurs at the liquid surface, portions of the liquid F
break free from the turbulent bulk liquid, resulting in
droplet formation. The dependence of atomization on
cavitation phenomena has been demonstrated for fre-
quencies between 0.5 and 2.0 MHz.[14,15] Boguslavskii E
and Eknadiosyants[16] combined these theories with- Fig. 2 Schematic diagram of a typical ultrasonic nebulizer.
their proposal that droplet formation resulted from (A) Face mask or mouthpiece; (B) baffles; (C) geyser of res-
capillary waves initiated and driven by cavitation piratory solution or suspension; (D) piezoelectric crystal; (E)
bubbles. internal fan; (F) battery or electrical source. (From Ref.[9].)
 3856 Ultrasonic Nebulizers

mass output and the particle size of the generated aero- Liquid Housing
sols are summarized in Table 1.
Whereas air-jet nebulizers are usually disposable or
sterilizable, ultrasonic nebulizers are too expensive to
be produced as disposable units and are thus used
repeatedly,[17] running the risk of bacterial contami-
nation.[18] Cleaning nebulizers and connecting tubing
is difficult, and the transfer of Gram-negative bacteria
between nebulization equipment and patients has been
reported[19] Mesh
Plezo electric element
Although most ultrasonic devices share a basic
design, some novel devices have been developed. The
Droplets
RespimatÕ (Boehringer Ingelheim) offered direct deliv-
ery of a metered dose from a valve similar to that used
in pMDIs to the surface of a vibrating crystal
(2.5 MHz). Although the mass median aerodynamic
diameter (MMAD) of th eaerosol generated was closed Fig. 3 Schematic diagram of the sprayhead of the Bespak
to 10 mm, generally considered too large for effective Piezo Electric Actuator. (Reproduced courtesy of Bespak
inhalation therapy, equivalence to an equivalent pMDI plc, UK.)
formulation was claimed.[20]
The Bespak Piezo Electric ActuatorÕ is a novel
aerosol delivery system based on a piezoelectric crystal
combined with an electroformed mesh (Fig.3). It pro- suspensions without requiring these to be formulated
duces droplets of ‘‘adjustable’’ size from a single with small suspended particles. Furthermore, they
metered drop or fluid reservoir.[21] The mesh hole can produce small droplets from large mesh holes.
dimension (as small as 3 mm) determines the size of For example, TouchSpray devices can deliver 5 mm
the droplets produced, whereas the size and density droplets of suspensions containing normally micro-
Trace–Ultrasonic

of the holes control the rate of fluid delivery. nized drug suspensions, which typically include parti-
These can be varied according to the formulation. cles of 3 mm or larger. These droplets can be
Although solutions are more readily nebulized, sus- produced by devices having mesh hole sizes of 15 mm.
pensions can be aerosolized if the particle size of the TouchSpray allows small droplets to be produced from
suspended particles is two to three times smaller than solutions. A review of the development of perforated
the mesh size. mesh devices is given in Humberstone, Sant, and
More recently, The Technology Partnership has cre- van Rensburg.[23] An inhalation droplet spray plume
ated several new forms of a perforated mesh atomizer produced by the TouchSpray is shown in Fig.4.
known as TouchSprayÕ.[22] TouchSpray devices deliver Future devices are likely to be hybrids of ultrasonic
nebulizer technology and pMDI or DPI technology.
Vibrating transducer or grids, incorporated into
pMDIs, could break down large propellant droplets
Table 1 Design features of ultrasonic nebulizers that would otherwise be wasted into a more respirable
determining particle size distribution and mass output size range. Alternatively, the dependence of DPIs
Design features Nebulizer characteristics on patient-generated turbulent airstreams could be
Piezoelectric crystal minimized by deaggregating powder by vibrational
Frequency of vibration elements within the inhalation device or by generating
Amplitude of vibration aerosols electrostatically (SpirosÕ, Dura).
Surface morphology
(flat or curved)
Coupling between AEROSOL DROPLET SIZE
crystal and fluid
Fluid reservoir
The efficacy of a therapeutic inhalation aerosol
Size depends onits ability to penetrate the respiratory tract.
Shape
This is primarily dependent on the particle size of the
Baffles
particles or droplets. To penetrate to the peripheral
Auxiliary air flow (respiratory) airways, aerosols generally require a size
Velocity less than approximately 5–6 mm, with a size less than
(Modified from Ref.[7].) 2 mm being optimal for alveolar deposition.[24,25]
Ultrasonic Nebulizers 3857

generating droplets of sizes comparable to arrow

air-jet devices. However, devices with low frequencies
(<0.5 MHz) tend to generate droplets outside the res-
pirable range.[4,32] Most commercially available ultra-
sonic nebulizers have an operating frequency between
1 and 2 MHz. Although these produce aerosol droplets,
which are significantly larger than those produced by
air-jet nebulizers,[3,4] the size distributions are generally
less polydisperse.[3,33]

NEBULIZATION TIME, DRUG OUTPUT,

AND RESIDUAL VOLUME

Patient compliance with prescribed nebulization

regimes is primarily determined by the duration of
the therapy. Nebulizer fluids may be atomized for a
set period, or more usually, a measured volume of
therapeutic liquid is nebulized to ‘‘dryness.’’ The time
taken to achieve this is directly related to the volume to
be delivered. However, not all the fluid in the nebulizer
can be atomized, and some fluid remains associated
with the baffles, internal structures, and walls of
Fig. 4 Inhalation droplet spray plume produced by Touch-
the nebulizer as the ‘‘dead’’ or ‘‘residual’’ volume.[34]
Spray device. (Reproduced courtesy of The Technology Part-
nership plc, UK.)
The proportion of fluid remaining as the residual
volume and thus unavailable to patients is higher for

Trace–Ultrasonic
smaller fill volumes.
Ultrasonic devices typically produce a much larger
fluid output per unit of time than do jet nebulizers.
Clinical performance is often predicted from in vitro Although Sterk et al.[4] suggested that high airflow
measurements of the droplet size of an aerosol. Nebu- through these devices resulted in lower droplet concen-
lized aerosols are usually assessed by a multistage trations per unit volume than for air-jet nebulizers,
liquid impinger (MSLI), cascade impactor, or laser dif- more recent findings indicate that the reduced volume
fraction methods. The MSLI and cascade impactors of dilution air in modern ultrasonic devices gives a
comprise a series of progressively finer jets and col- more concentrated aerosol cloud. Ultrasonic nebuli-
lection plates, permitting fractionation of aerosols zers may retain a higher residual volume than do com-
according to their MMADs.[26] Problems associated parable air-jet nebulizers,[35] but they show less
with the cascade impactors for the assessment of neb- tendency to increase the solute concentration within
ulized aerosols are that the high flow rates involved the nebulizer chamber during operation.[27,29]
(typically 28–90 L/min) give rise to rapid solvent evap- In comparing DeVilbiss ultrasonic and jet devices,
oration and droplet entrainment. Additionally, these Newman, Pellow, and Clarke[29] reported shorter neb-
inertial impaction techniques are invasive, laborious, ulization times for the ultrasonic devices with higher
and time-consuming. Alternatively, nebulized aerosols fluid outputs. Furthermore, the air-jet devices tended
may be sized by passing the spray through a beam of to increase the concentration of drug in the nebulizer
a laser diffraction analyzer. The volume or mass chamber more than did the ultrasonic counterparts.
median diameter is then calculated from the generated The output from nebulizers comprises droplets of
diffraction pattern. Provided the liquid density is drug solutions and suspensions and solvent vapor,
known, the MMAD can be calculated. which saturates the outgoing air, causing solute con-
Numerous investigators have compared the aerosol centration to increase during nebulization. Because
droplet size of nebulized aerosols from ultrasonic ultrasonic nebulizers generally have higher fluid out-
and air-jet devices.[3,4,6,27–31] Because droplet size is puts and larger droplet sizes than do air-jet nebulizers,
inversely proportional to the acoustic frequency, there is more solvent available within the dense aerosol
smaller droplets are generated from ultrasonic devices cloud to saturate the outgoing air. Thus, the changes in
with higher frequencies. Ultrasonic nebulizers with solute concentration within the dead volume are much
high operating frequencies (2–3 MHz) are capable of smaller than for air-jet devices.
 3858 Ultrasonic Nebulizers

THERMAL EFFECTS properties (density, surface tension, viscosity) in

conjunction with the nebulizer design and operating
Excess energy within the nebulizer is converted to heat, conditions. Empirical and semiempirical formulas pre-
causing the temperature of the liquid in the reservoir to dict that the droplet size of the aerosols is proportional
increase until the input energy balances the energy to surface tension and inversely related to viscosity,
removed by evaporation and by conduction to the sur- whereas the effect of density, over the concentration
roundings and circulating air.[12] The temperature of a range normally encountered, is negligible. The filtering
liquid in an ultrasonic nebulizer may thus increase by effects of the baffles and solvent evaporation may
as much as 20
C above ambient temperature during modify the secondary aerosol produced, but stu-
use.[6,36] This results in changes in the properties of dies[32,37,45,46] have found viscosity to be a major
the fluid, including surface tension and viscosity. determinant of aerosol size and output characteristics.
Although such changes may affect the aerosol particle High-viscosity fluids offer greater resistance to the
size characteristics and overall drug output, little vari- integral fountain-disintegration process, thereby pro-
ation was seen during prolonged operation of Medix ducing not only lower output but also larger droplets.
Electronic or Easimist nebulizers[37] or when the tem- Gershenzon and Eknadiosyants[14] noted higher
perature in a DeVilbiss Aerosonic nebulizer was droplet outputs with lower viscosity fluids, whereas
increased using a heating coil.[38] Boucher and Kreuter[32] reported that it was difficult
Warming may have a beneficial effect. For instance, to ultrasonically aerosolize fluids with viscosities
the temperature of fluids atomized in air-jet nebulizers exceeding 10 cP. Gershenzon and Eknadiosyants[14]
decreases by approximately 10–15
C during use,[34,39] stated that the atomization rate for a wide range of
resulting in bronchoconstriction in some asthma suf- fluids (except water) is given by the proportionality
fers.[40] Bronchoconstriction, which is most marked at of A2 to pr/Zs, where A is the atomization rate, r is
5
C, disappears at 37
C and thus may be minimized the liquid vapor pressure, Z is the viscosity, and s is
by using an ultrasonic device. Furthermore, when the surface tension.
solutions of drugs with low solubility are to be nebu- Il’in and Eknadiosyants[15] suggested that the
lized, ultrasonic nebulizers, which warm the solutions, dynamic viscosity coefficient was the most important
may be preferable[36] to air-jet devices, which cool them property determining the nebulization rate. The rate
Trace–Ultrasonic

and may cause precipitation.[39] However, the heat of nebulization for solutions of tyloxapol or N-
generated may harm heat-labile materials such as acetyl-L-cysteine decreased progressively as the solution
diethylenetriaminepentoacetic acid (99mTc-/DTPA),[41] viscosity increased. When certain oily and viscous liquids
proteins,[31] and some antibiotic solutions.[42] Thus, (e.g., LipiodalÕ; a radio-opaque diagnostic agent) were
ultrasonic nebulizers are specifically prohibited for aero- nebulized, a fountain of liquid was generated, although
solization of recombinant human deoxyribonuclease this did not disintegrate to produce an aerosol. Similar
(rhDNase).[1] results were reported by McCallion et al.[37], who com-
pared a range of different model fluids, finding that
ultrasonic nebulizers could not efficiently atomize the
FORMULATION OF NEBULIZER FLUIDS more viscous liquids and tended to produce poor total
fluid outputs. Furthermore, when the fluid viscosity
Respiratory solutions are the mainstay of nebulized increased, larger droplets were generated.
inhalation therapy. These typically contain drug dis- Surface tension is also important because it repre-
solved in aqueous, isotonic solvent systems, with the sents the force resisting the formation of new surfaces.
inclusion of cosolvents (e.g., ethanol) if necessary. Sus- These forces tend to impair atomization by opposing
pensions are less common for nebulization, although any distortion or irregularity on the liquid surface,
corticosteroid preparations such as budesonide are thereby delaying the onset of fountain formation. In
available. Antioxidant and antimicrobial preservatives suspension formulations, surfactants may be present
may be included in the formulations; however, some as suspending agents. Reduction in liquid surface ten-
(e.g., sodium metabisulfate, benzalkonium chloride, sion, through the addition of these agents, may
EDTA) may have paradoxical effects and cause cough- decrease nebulization rate. This may be attributed to
ing and bronchoconstriction.[43] To prevent this, reduction in capillary wavelength, causing an increase
‘‘preservative-free’’ unit dose products are marketed. in threshold amplitude[45] or through their influence
Isotonicity is generally achieved using sodium chloride. on the diffusion of gas into cavitation bubbles.[47]
Although iso-osmotic solutions of pH 3–8.5 are usually When a range of pharmaceutically relevant surfactant
employed, the osmolarity and pH may change during systems was nebulized, an inverse relationship was
use, resulting in bronchospasm.[44] found between droplet size and surface tension over
The size and output characteristics of aerosols gener- the entire concentration range investigated or to a peak
ated from such liquids depend on their physicochemical value.[33] There was no relationship between this peak
Ultrasonic Nebulizers 3859

value and a specific surface tension or the critical ultrasonic nebulizers for the delivery of recombinant
micelle concentration. In most cases, the total fluid consensus a-interferon. The extent of protein aggre-
output was unchanged. gation was governed by the type of nebulizer used
Most nebulizer formulations are solutions, but a few with, the Easimist causing the least and Microstat the
corticosteroid suspension formulations have been mar- most aggregation. This was related to the increase in
keted. In general, ultrasonic nebulizers are less efficient temperature and could be minimized by cooling the
and more variable in delivering suspensions than are nebulizer solution during use. The AerosonicÕ nebuli-
air-jet nebulizers. Although soluble radiopharmaceuti- zer completely inactivated lactate dehydrogenase after
cals may be more appropriate for delivery from ultra- 20 min.[38] The profile of inactivation differed from that
sonic nebulizers,[48] Lin et al.[49] successfully used an of an air-jet nebulizer and was associated both with the
ultrasonic nebulizer to deliver radiolabeled sulfur and temperature increase of the nebulizer fluid and the
tin colloids for lung imaging. McCallion et al.[50] neb- aerosol production. The activity of the enzyme was
ulized a range of latex sphere suspensions in air-jet almost completely retained if Tween 80 or PEG 8000
and ultrasonic nebulizers. No correlation was found was included in the nebulizer fluid.
between the size of the suspended spheres and the size
distribution of the nebulized droplets. Higher outputs
of smaller spheres were reported, with a concentrating GUIDELINES AND STANDARDIZATION
effect occurring in the residual volume. The ultrasonic FOR THE USE OF NEBULIZERS
nebulizer studied was less efficient than the jet nebuli-
zers, degrading some of the larger spheres and The British Thoracic Society Nebulizer Project Group
being unable to atomize suspended spheres of a specific published a set of guidelines for nebulizer treatment.[18]
size range. Their findings relating to ultrasonic devices are sum-
Nebulizers, particularly air-jet devices, have been marized as follows:
studied extensively for the delivery of liposomes to
the lung (e.g., Refs.[51,52]). Because in ultrasonic nebu- 1. The aim of treatment is to deliver a therapeutic
lizers, thetemperature of fluid in the reservoir is raised dose of the drug as an aerosol in the form of res-
during use, they have generally been avoided for deli- pirable particles within 5–10 min.

Trace–Ultrasonic
vering liposomes, which exhibit temperature-depen- 2. Nebulizers are useful when large doses of drug
dent drug release. Barber and Shek[53] reported that are to be inhaled by patients too ill to use alter-
egg phosphatidylcholine (egg PC) liposomes with a native devices and when drugs are not available
mean size of 281 nm or smaller were stable to nebuliza- for delivery from pMDIs and DPIs.
tion in a DeVilbiss Ultra-Neb 99Õ ultrasonic nebulizer. 3. Nebulizers are generally used to treat acute
However, dipalmitoylphosphatidylcholine liposomes exacerbations of asthma or chronic obstructive
of 499 nm increased in size within the nebulizer reser- pulmonary disease. Other indications include
voir, suggesting fusion of vesicles, which could result long-term bronchodilator treatment of chronic
in loss of entrapped hydrophilic materials. A later airflow obstruction; prophylactic treatment for
study[54] showed that the size of large multilamellar asthma; antimicrobial drugs for cystic fibrosis,
egg PC liposomes remaining in a Medix Electronic bronchiectasis, and HIV/AIDS; and sympto-
nebulizer decreased markedly during nebulization, matic relief in palliative care.
suggesting vesicle disruption, which was reduced by 4. When drugs other than bronchodilators are
including cholesterol in the formulations. being nebulized, equipment known to provide
Ultrasonic nebulizers are less suitable than jet nebu- a suitable output should be used, and specific
lizers for delivery of proteins to the airways because of instructions should be given to patients. Such
their thermal sensitivity. In a comparison of eight air- treatment should be supervised by hospital
jet and two ultrasonic nebulizers, all air-jet nebulizers specialists.
maintained the enzymatic activity of rhDNase in both 5. Nebulization times for bronchodilators should
the collected aerosol and the residual volume.[31] With be less than 10 min. Nebulization times and cor-
the ultrasonic nebulizers, some thermal denaturation rect operation of devices should be familiar to
of the enzyme was evident toward the end of the neb- the patients.
ulization period when the liquid volume was minimal
and its temperature highest. The maximum tempera- Convenience and patient preference determine
ture of the rhDNase solution was 58
C, which was whether a mouthpiece or face mask is used.
near the thermal transition temperature (approxi- Currently, there are no U.S. Pharmacopeia or
mately 65
C) of the enzymes.[31] European Pharmacopoeia standard tests for charac-
Ip et al.[55] investigated DeVilbiss AerosonicÕ, terizing the output of respiratory solutions or suspen-
Mountain Medical MicrostatÕ, and Medix ElectronicÕ sions generated by air-jet or ultrasonic nebulizers.
 3860 Ultrasonic Nebulizers

If nebulizers are to truly compete with pMDIs and 22. Humberstone, V.C.; Newcombe, G.C.F.; Sant, A.J.;
DPIs, standardization of methods to measure dead Palmer, M.R. Fluid droplet production apparatus and
method. EP-615-470. 1994.
volume, quantity of aerosol emitted from the device, 23. Humberstone, V.C.; Sant, A.J.; van Rensburg, R.J. Piezo-
nebulization times, and the size distribution of nebu- electric Aerosol Inhalers: Technology to Product, Confer-
lized clouds is a logical and fundamental requirement. ence Proceedings of New Horizons in Pulmonary Drug
Delivery, October, 1–2, 1996.
24. Stahlhofen, W.; Gebhart, J.; Heyder, J. Experimental deter-
mination of the regional deposition of aerosol particles in
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Trace–Ultrasonic
 Unit Processes in Pharmacy: Fundamentals
Anthony J. Hickey
Department of Pharmaceutics, The University of North Carolina at Chapel Hill,
Chapel Hill, North Carolina, U.S.A.

David Ganderton
Devon, U.K.

INTRODUCTION the body is m and its velocity u, the kinetic energy is

1/2 mu2, and for unit mass of fluid the kinetic energy
Pharmaceutical manufacturing can be divided into a is u2/2.
number of unit process on the basis of a few funda- The total mechanical energy of unit mass of fluid is
mental principles. The following monograph describes therefore
briefly fluid flow and heat and mass transfer.
u2 P
þ þ zg
2 r
FLUID FLOW
and proved no energy is lost or gained by the system,
Fluids (liquids and gases) are a form of matter that the mechanical energy at two points A and B is the
cannot achieve equilibrium under an applied shear same as that expressed by Eq. (1).
stress but deform continuously, or flow, as long as
u2A PA u2 PB
shear stress is applied. þ þ zA g ¼ B þ þ zB g ð1Þ
Streamlines are hypothetical lines without width 2 r 2 r
drawn parallel to all points to the motion of the fluid. This relationship neglects the frictional degradation
As velocity increases, pressure decreases. Pressure field of mechanical energy that occurs in real systems. A
around an object is the reverse of velocity field. This fraction of the total energy is dissipated in overcoming
may appear to contradict common experience. How- shear stresses induced by velocity gradients in the fluid.
ever, it follows from the principle of conservation of Energy E lost during flow between A and B must be
energy and finds expression in Bernoulli’s theorem. considered. The dimensions of each component are
Unit–Validation

L2 T
2, Length2 Time
2. In practice, each term is

divided by g (LT
2) to give the dimension of length.
Bernoulli’s Theorem
The terms are then referred to as velocity head, press-
ure head, potential head, and friction head, the sum
At any point in system through which a fluid is flow-
giving the total head of the fluid as shown in Eq. (2).
ing, the total mechanical energy can be expressed in
terms of potential energy, pressure energy, and kinetic u2A PA u2 PB E
energy. þ ¼ B þ þ zB þ ð2Þ
2g rg þ zA 2g rg g
Potential energy of a body is its capacity to do work
by reason of its position relative to some center of The evaluation of the kinetic energy term requires
attraction. For unit mass of fluid at a height z above consideration of the variation in velocity found in the
some reference level, the potential energy is zg. direction normal to flow. The kinetic energy, given
Pressure energy or flow energy is an energy form by the term u2mean/2, differs from the true kinetic
peculiar to the flow of fluids. The work done and the energy found by summation across the flow direction.
energy acquired in transferring the fluid is the product The former can be retained, however, if a correction
of the pressure P and the volume. factor a is introduced. Then
Volume of unit mass of the fluid is the reciprocal of
the density r. For an incompressible fluid, the density u2mean
velocity head ¼
is not dependent on the pressure, so for unit mass of 2ga
fluid the pressure energy is P/r.
Kinetic energy is a form of energy possessed by a where a has a value of 0.5 in laminar flow and
body by reason of its movement. If the mass of approaches unity in fully turbulent flow. Mechanical
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-100001925
3862 Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved.
Unit Processes in Pharmacy: Fundamentals 3863

energy is added to the system at some point by means known as the coefficient of discharge, as shown in
of a pump. The work W done, in absolute units, on a Eq. (6).
unit mass of fluid at A is given by Eq. (3).
vffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
W u2 PA u2 PB E u 2ðP
P Þ
u 1 2
þ A þ þ zA ¼ B þ þ Q ¼ CD a2 t   ð6Þ
g 2g pg 2g rg þ zB g p 1
a2 =a21 2
ð3Þ
The power required through a system at a certain The value of CD depends on conditions of flow and
rate may be calculated using Eq. (3) to drive a liquid. shape of the constriction. For a well-shaped constriction
The sum of heads, DH, being the total head against (notably circular cross section), it would vary between
which the pump must work is therefore 0.95 and 0.99 for turbulent flow. The value is much lower
in laminar flow because the kinetic energy correction is
W
þ DH larger. The return of the fluid to the original velocity
g by means of a diverging section forms a flow-measuring
device known as a Venturi meter.
If the work performed and energy acquired by unit The Venturi meter is shown in Fig. 1A. The con-
mass of fluid is DHg, the power required to transfer verging cone leads to the narrowest cross section
mass m in time t is given by known as the throat. The change in pressure is mea-
sured across this part of the meter and the volumetric
DHgm
Power ¼ flow rate found by substitution into Eq. (6). Values of
t the coefficient of discharge are given previously. The
diverging section or diffuser is designed to induce a
As the volume flowing in unit time Q is m/rt, the
gradual return to the original velocity. This minimizes
power is given by Eq. (4).
eddy formation in the diffuser and permits the recovery
of a large proportion of the increased kinetic energy as
Power ¼ QDHgr ð4Þ
pressure energy. The permanent loss of head due to
friction in both converging and diverging sections is
small. The meter is therefore efficient.
Flow Measurement When the minimization of energy degradation is less
important, the gradual economic return to the original
The Bernoulli theorem can also be applied to the velocity may be abandoned, compensation for loss of
measurement of flow rate. The passage of an incom- efficiency being found in a device that is simpler,
pressible fluid through a constriction results in an cheaper, and more adaptable than the Venturi meter.

Unit–Validation
increase in velocity from u1 to u2, which is associated The orifice meter, to which this statement applies, con-
with a decrease in pressure from P1 to P2, which can sists simply of a plate with an orifice. A representation
be measured directly. The volumetric flow rate Q ¼ of flow through the meter is depicted in Fig. 1B,
u1a1 ¼ u2a2 by algebraic rearrangement (which is not indicating convergence of the fluid stream after passage
shown). The final linear velocity u2 can be described through the orifice to give across section of minimum
by Eq. (5). area called the vena contracta. The downstream
pressure tapping is made at this cross section. The
u22 u2 P1
P2 volumetric flow rate is given by Eq. (6) in which a2 is

1 ¼ ð5Þ
2 2 p the jet area at the vena contracta. The measurement
of this dimension is inconvenient. It is therefore related
The volumetric flow rate can be described by to the area of the orifice, a0, which can be accurately
Eq. (6). measured by the coefficient of contraction Cc by the
vffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi relation Cc ¼ a2/a0.
u 2ðP
P Þ The coefficient of contraction, frictional losses
u 1 2
Q ¼ a2 t   between the tapping points, and kinetic energy cor-
p 1
a2 =a21 2
rections are absorbed in the coefficient of discharge.
The volumetric flow rate is then given by Eq. (7).
This derivation neglects the correction of kinetic
energy loss due to nonuniformity of flow in both cross vffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
u 2ðP
P Þ
sections and the frictional degradation of energy dur- u 1 2
Q ¼ CD a0 t   ð7Þ
ing passage through the constriction. This is corrected p 1
a20 =a21
by the introduction of a numerical coefficient, CD,
 3864 Unit Processes in Pharmacy: Fundamentals

A B

To manometers Pressure Pressure

tapping tapping

Vena
contracta
Onfince
C D

Us B

Pa Pb

Fig. 1 (A) Venturi meter; (B) orifice meter; (C) Pitot tube; and (D) rotameter.

If the orifice is small compared with the pipe cross As the velocity at B is zero, Eq. (10) reduces to
section, the term (1
(a20 /a21)) approaches unity. As
P1
P2 ¼ Dhrg, Dh being the difference in head u2a Pb
Pa
developed by the orifice, Eq. (7) reduces to Eq. (8). ¼
2 r

Q ¼ CD a0 ðDhrgÞ1=2 ð8Þ and u2 can be calculated. As only a local velocity is

measured, variation of velocity in a section can be
Unit–Validation

studied by altering the position of the tube. This pro-

The value of CD for the orifice meter is about 0.6, cedure must be used if the flow rate in a pipe is to be
varying with construction, the ratio a0/a1, and flow measured. The mean velocity is derived from velocities
conditions within the meter. Due to its complexity, it measured at different distances from the wall. This
cannot be calculated. derivation and the low pressure differential developed
After passage through the orifice, flow disturbance render the Pitot tube less accurate than either the ven-
during retardation causes the dissipation of most of turi tube or the orifice meter for flow measurement.
the excess kinetic energy as heat. The permanent loss Hence, the tube is small in comparison with the pipe
of head is therefore high, increasing as the ratio of diameter and, therefore, produces no appreciable loss
a0/a1 falls, ultimately reaching the differential head of head.
produced within the meter. The rotameter (a variable area meter), shown in Fig.
The Bernoulli theorem may be used to determine 1D, is commonly used, giving a direct flow rate reading
the change in pressure caused by retardation of fluid by the position of a small float in a vertical, calibrated
at the upstream side of a body immersed in a fluid glass tube through which the fluid is flowing. The tube
stream. This principle is applied in the Pitot tube, is internally tapered toward the lower end so that the
shown in Fig. 1C. The fluid velocity is reduced annulus between float and wall varies with the position
from ua, the velocity of the fluid filament in alignment of the float. Acceleration of the fluid through the annu-
with the tube, to zero at B, an position known as the lus produces a pressure differential across the position
stagnation point. The pressure, Pb, is measured at this of the float and an upward force upon it. At the equi-
point by the method shown in Fig.1 C. The undis- librium position, which may be stabilized by a slow
turbed pressure, Pa, is measured in this example with rotation of the float, this upward force is balanced by
a tapping point in the wall connected to a manometer. the weight force acting on the float. If the equilibrium
Unit Processes in Pharmacy: Fundamentals 3865

is disturbed by increasing the flow rate, the balance of and assesses the additional effects of pipe roughness,
weight force and the pressure differential are produced changes in diameter, bends, exists, and entrances.
by movement of the float upward to a position at When the total pressure drop due to friction is known
which the area of the annulus is bigger. For accurate for the system, the equivalent head can be derived and
measurement, the rotameter is calibrated with the fluid the power requirement for driving a liquid through the
to be metered. Its use is, however, restricted to that system can be calculated from Eq. (9).
fluid.
Streamline flow in a tube
Laminar and Turbulent Flow
The mathematical analysis of streamline flow in a sim-
The nature of flow may be examined by introducing a ple tube results in the expression known as Poiseuille’s
dye into the axis of the tube. At low speeds, the dye law, one form of which is Eq. (9).
forms a coherent thread, which grows very little in
thickness with distance down the tube. However, with
progressive increase in speed, the line of dye begin to DPpd4
Q ¼ ð9Þ
waver and then break up. Secondary motions crossing 128Zl
and recrossing the general flow direction occur.
Finally, at very high speeds, no filament of dye is where Q is the volumetric flow rate or discharge, DP is
detected and mixing is instantaneous. In this experi- the pressure drop across the tube, d and l are the diam-
ment, flow changes from laminar to turbulent, the eter and length of the tube, respectively, and Z is the
change occurring at a critical speed. Generalizing, in viscosity of the fluid.
laminar flow, the instantaneous velocity at a point is Where flow in the tube is streamline or turbulent, an
always the same as the mean velocity in both magni- infinitesimally thin stationary layer is found at the wall.
tude and direction. In turbulent flow, order is lost The velocity increases from zero at this point to a
and irregular motions are imposed upon the main maximum at the axis of the tube. The velocity profile
steady motion of the fluid. At any instant of time, of streamline flow is shown in Fig. 9A. The velocity
the fluid velocity at a point varies in both magnitude gradient du/dr varies from a maximum at the wall to
and direction, having components perpendicular as zero at the axis. In flow through a tube, the rate of
well as parallel to the direction of net flow. Over a per- shear is equal to the velocity gradient, and Eq. (1) dic-
iod of time, these fluctuations even out to give the net tates the same variation of shear stress.
velocity in the direction of flow. To derive Poiseuille’s law, the form of the velocity
In turbulent flow, rapidly fluctuating velocities profile must first be established. For a fluid contained
produce high velocity gradients within the fluid. within a radius r flowing in a tube of radius R, the
Proportionately large shear stresses are developed, pressure drop across length l is DP; therefore, the

Unit–Validation
and to overcome them, mechanical energy is degraded pressure force driving this section is DPpr2. If the flow
and dissipated in the form of heat. The degradation of is steady, this force can only be balanced by opposing
energy in laminar flow is much smaller. viscous forces acting on the ‘‘wall’’ of the section. This
The turbulent mechanism that carries motion, heat, force is
or matter from one part of the fluid to another is
absent in laminar flow. The agency of momentum
transfer is the shear stress arising from the variations DPr
t ¼
in velocity, that is, the viscosity. Similarly, heat and 2l
matter can only be transferred across streamlines on
a molecular scale, heat by conduction and matter by
Substituting for Eq. (1) gives
diffusion. These mechanisms that are present but less
important in turbulent flow are comparatively slow.
Velocity, temperature, and concentration gradients du DPr

¼
are, therefore, much higher than in turbulent flow. dr 2Zl

The Flow of Liquids in Pipes The velocity gradient is negative because u decreases as
r increases. If r ¼ R, then u ¼ 0. Integration gives
The many pharmaceutical processes that involve the
transfer of a liquid confer great importance on the Z u Z r
study of flow in pipes. This study permits the evalu- DP
du ¼ r dr
ation of pressure loss due to friction in a simple pipe 0 2Zl R
 3866 Unit Processes in Pharmacy: Fundamentals

Therefore, Eq. (10) can be written as 0.1

 
DP R2
r 2
u ¼ ð10Þ Transition Turbulent flow
2Zl 2 region

R/pu2
0.01 Rough pipe (e/a=0.05)

This relation shows that the velocity distribution Rough pipe (e/a=0.005)
Laminar
across the tube is parabolic. For such a distribution, flow Smoo
th pip
the maximum velocity is twice the mean velocity. e
The volumetric flow rate across an annular section 0.001
102 103 104 105 106
between r and (r þ dr) is given by
Re=(pud)/η
Q ¼ 2pr dr r: Fig. 2 Pipe friction chart R/ru2 vs. Re.

Substituting for u for Eq. (10) gives

pipe area completes the data required for calculating
DPp 2 Re. If the pipe roughness factor is known, the equiva-
Q ¼ ðR r
r 2 Þr dr lent value of R/ru2 can be read from Fig. 2, and the
2Zl
shear stress at the pipe wall calculated. The total fric-
The total volumetric flow rate is the integral between tional force opposing motion is the product of R and
the limits r ¼ R and r ¼ 0: the surface area of the pipe, pdl, where l is the length
of the pipe. If the unknown pressure drop across the
Z R pipe is DP, the force driving through the pipe is
DPp
Q ¼ ðR2 r
r 3 Þdr DPpd2/4. Equating pressure force and frictional force,
2Zl 0 !R
r2 4
DPp 2
r
¼ R 2 pd2
2Zl 4 DP ¼ ¼ Rpdl
0 4

Therefore Eq. (9) is valid: Therefore, Eq. (12) follows.

DPpR4 DPpd4 4Rl2

Q ¼ ¼ DP ¼ ð12Þ
8Zl 128Zl d

where d is the diameter of the tube. Division by rg gives the pressure loss as a friction
As Q ¼ umeanp(d2/4), substitution and rearrange- head.
Unit–Validation

ment gives Eq. (11).

32umean Zl
DP ¼ ð11Þ Flow in Tubes of Non-Circular Cross Section
d2

The foregoing arguments may be applied to turbulent

flow in noncircular ducts by introducing a dimension
The Significance of Reynolds Number Re equivalent to the diameter of a circular pipe. This is
known as the mean hydraulic diameter, dm, which is
The Reynolds number describes the ratio of the inertia defined as four times the cross-sectional area divided
and viscous or frictional forces. Higher the Reynolds by the wetted perimeter. The following examples are
number, greater is the relative contribution of inertial given:
effects. At very low Re, viscous effects predominate For a square channel of side b,
and the contribution of inertial forces can be ignored.
4b2
dm ¼ ¼ b
4b
Calculation of the Pressure Drop
in a Pipe Due to Friction
For an annulus of outer radius r1 and inner radius r2,
If the volumetric flow rate of a liquid of density r and
viscosity Z through a pipe of diameter d is Q, the deri- 4ðpr21
pr22 Þ
¼ 2ðr1
r2 Þ
vation of the mean velocity u from the flow rate and 2pr1 þ 2pr2
Unit Processes in Pharmacy: Fundamentals 3867

This simple modification does not apply to laminar

flow in noncircular ducts.
103

Frictional Losses at Pipe Fittings

R′/pu2
In addition to the friction losses at the wall of a 101
straight pipe, losses occur at the various fittings and
valves used in practical systems. In general, these losses
are derived from sudden changes in the magnitude or
direction of flow induced by changes in geometry. They
10-1
can be classified as losses due to a sudden contraction
of enlargement, losses at entrance or exit, and losses
due to pipe curvature. Losses can be conveniently
10-3 10-1 101 103
expressed as a length of straight pipe offering the same
Re′= (pud)/η
resistance. This is usually in the form of a number of
pipe diameters. For example, the loss at a right-angled Fig. 3 R0 /ru2 vs. Re0 number for a smooth sphere.
elbow is equivalent to the length of a straight pipe
equal to 40 diameters. The sum of the equivalent
lengths of all fittings and vales is then added to the Therefore,
actual pipe length and the total frictional loss
estimated by Eq. (20). pd2 12 pd2
Total drag force ¼ R ¼ ru2 ¼ 3pZdu
4 Re 4
ð14Þ
Motion of Bodies in a Fluid
This is the normal form of Stokes law.
Viscous and inertial forces operate to determine the At larger Re0 values, the experimental curve pro-
flow pattern and drag force on a body moving relative gressively diverges from this relation, ultimately
to a fluid. The Reynolds number, which expresses their becoming independent of Re0 and giving a value of
ratio, is used as a parameter to predict flow behavior. R/ru2 equal to about 0.22. As Re0 increases, the form
The relation between the drag force and its controlling drag increases, ultimately becoming solely responsible
variables is presented in a manner similar to that for the force opposing motion.
employed for flow in a pipe. Considering a sphere For nonspherical particles, the analysis employs the
diameter of a sphere of equivalent volume. A correc-

Unit–Validation
moving relative to a fluid, the projected area normal
to flow is pd2/4, where d is the diameter of the sphere. tion factor, which depends upon the shape of the body
The drag force acting on the unit projected area R0 is and its orientation in the fluid, must be applied.
determined by the velocity u, the viscosity Z, and the An important application of this analysis is the esti-
density r of the fluid and the diameter of the sphere mation of the speed at which particles settle in a fluid.
d. Dimensional analysis yields Eq. (13). Under the action of gravity, the particle accelerates
until the weight force mg is exactly balanced by the
  opposing drag. The body then falls at a constant ter-
R udr
¼ fðReÞ ¼ f ð13Þ minal velocity u. Equating weight and drag forces gives
ru2 Z
Eq. (15).
This form of Reynolds number, Re0 , employs the
diameter of the sphere as the linear dimensions. p 3 pd2
mg ¼ d ðrs
rÞg ¼ R ð15Þ
With the exception of an analysis at very low Re. 6 4
values, the form of this function is established by
experiment. Results are presented on logarithmic where r is the density of the particle.
coordinates in Fig. 3. When Re0 is less than about For a sphere falling under streamline conditions
0.2, viscous forces are solely responsible for drag on (Re0 0.2), R0 ¼ ru2 (12/Re0 ). Substituting in Eq. (15)
the sphere and Eq. (13) becomes gives Eq. (16).

R 12 d2 ðrs
rÞg
2
¼ u ¼ ð16Þ
ru Re 18Z
 3868 Unit Processes in Pharmacy: Fundamentals

This expression follows more simply from the equation through a bed of the same material with a porosity of
mg
3pdZu. 25%. It would also flow more quickly through a bed
of coarse particles than through a bed of fine particles
packed to the void fraction or porosity. The latter effect
Flow of Fluids through Packed Beds can be expressed in terms of the surface area offered to
the fluid by the bed. This property is inversely pro-
The analysis of the flow of fluids through a permeable portional to the size of the particles forming the bed.
bed of solids is widely applied in filtration, leaching, Permeability increases as the porosity increases and
and several other processes. A first approach may be the total surface of the bed decreases and these factors
made by assuming that the interstices of the bed corre- may be combined to give the hydraulic diameter d0 of
spond to a large number of discrete, parallel capillaries. an equivalent channel. This is defined by:
If the flow is streamline, the volumetric flow rate Q is
given for a single capillary by Eq. (14): Volume of voids
d ¼
Total surface of material forming bed
DPpd4
Q ¼
128Zl
The volume of voids is the porosity, and the volume
where l is the length of the capillary and d its diameter, of solids is (1
e). If the specific surface area, that is,
DP is the pressure drop across the capillary, and Z is the surface area of unit volume of solids, is S0, the total
the viscosity of the fluid. The length of the capillary surface presented by unit volume of the bed is S0(1
e).
exceeds the depth of the bed by a value that depends Therefore, Eq. (18) applies.
upon its tortuosity. The depth of bed, L, is however,
proportional to the capillary length l so that e
d ¼ ð18Þ
S0 ð1
eÞ
DPd4
Q ¼ ;
kZL Under streamline conditions, the rate at which a
fluid flows through this equivalent channel is given
where k is a constant for a particular bed. If the area by Poiseuille’s Eq. (14).
of the bed is A and it contains n capillaries per unit The velocity u’ in the channel is derived by dividing
area, the total flow rate is given by the volumetric flow rate by the area of the channel,
k0 d0 2. Combining the constants,
DPd4 nA
Q ¼
kZL
Q DPd2
Unit–Validation

u ¼ 2
¼
Both n and d are not normally known. However, they kd kZL
have certain values for a given bed, expressed by
Eq. (17).
This velocity, when averaged over the entire area of the
bed, solids, and voids, gives the lower value u. These
DP
Q ¼ KA ð17Þ velocities are related by
ZL

where K ¼ d4n/k. This constant is a permeability coef- u ¼ ue

ficient and its reciprocal, 1/K, is the specific resistance.
Its value characterizes a particular bed.
Therefore:
The postulate of discrete capillaries precludes valid
comment on the factors that determine the permea-
bility coefficient. Channels are not discrete but are u DPd2
interconnected in a random manner. Nevertheless, the ¼
e kZL
resistance to the passage of fluid must depend on
the number and dimensions of the channels. These
can be expressed in terms of the fraction of the bed Substituting for d0 by means of Eq. (18) gives
which is void, that is, the porosity, and the manner in
which the void fraction is distributed. Illustrating this
u DP e2
reference to a specific example, water would flow more ¼
easily through a bed with a porosity of 40% than e kZL ð1
eÞ2 S02
Unit Processes in Pharmacy: Fundamentals 3869

and Eq. (19) pharmacy. Heat energy can only be transferred from a
region of higher to a region of lower temperature.
DP e3 Understanding heat transfer requires the study of the
u ¼ ð19Þ mechanism and rate of this process.
kZL ð1
eÞ2 S02
Heat is transferred by three mechanisms: conduc-
tion, convection, and radiation. It is unusual for the
In this equation, known after its originator as Kozeny’s transfer to take place by one mechanism only.
equation, the constant k00 has a value of 5  0.5. As Q ¼ Conduction is the most widely understood
uA, where A is the area of the bed, Eq. (19) can be mechanism of heat transfer and the main method in
transformed to Eq. (20). solids. The flow of heat depends upon the transfer of
vibrational energy from one molecule to another and,
DPA e3 in the case of metals, the movement of free electrons.
Q ¼  ð20Þ Radiation is rare in solids but examples are found
ZL 5ð1
eÞ2 S02
among glasses and plastics. Convection by definition,
is not possible under these conditions. Conduction in
This analysis shows that permeability is a complex func- the bulk of fluids is normally overshadowed by
tion of porosity and surface area, the latter being convection, but it assumes great importance at fluid
determined by the size distribution and shape of the boundaries.
particles. The appearance of specific surface in Eq. (20) Heat is transferred from or to a region by the
offers a method for its measurement and provides the motion of fluids and the phenomenon of convection.
basis of fluid permeation methods of size analysis. This In natural convection, the movement is caused by
equation also applies in the studies of filtration. buoyancy forces induced by variations in the density
of the fluid; these variations are caused by differences
in temperature. In forced convection, movement is
Pumps
created by an external agency such as a pump.
All bodies with a temperature above absolute zero
Eqs. (8) and (9) examine the power requirement for
radiate heat in the form of electromagnetic waves.
driving a liquid through a system against an opposing
The radiation may be transmitted, reflected, or
head. This energy is normally generated by a pump. In
absorbed by matter, the fraction absorbed being trans-
different processes, the quantities to be delivered, the
formed into heat. Radiation is of great importance at
opposing head, and the nature of the fluid vary widely,
very high or very low temperatures and under circum-
and many pumps are made to meet these differing stances in which the other modes of heat transmission
requirements. Basically, however, pumps can be div-
are suppressed. Although the heat losses can, in some
ided into positive-displacement pumps, which may be
cases, equal the losses by natural convection, the mech-
reciprocating or rotary, and impeller pumps. Positive-

Unit–Validation
anism is, from the standpoint of pharmaceutical pro-
displacement pumps seek to displace a fixed volume
cessing, least important and needs only brief
of fluid with each stroke or revolution; impeller pumps,
consideration.
on the other hand, impart high kinetic energy to the
Heat transfer in many systems occurs as a steady-
fluid which is subsequently converted to pressure
state process, and the temperature at any point in the
energy. The volume discharged depends upon the system does not vary with time. In other important
opposing head.
processes, temperatures in the system do vary with
The equipment for pumping gases and liquids is
time. This situation, which is common among the
essentially similar. Machines delivering gases are
small-scale, batch-operated processes of the pharma-
commonly called compressors or blowers; compressors
ceutical and fine chemical industry, is known as
discharge at relatively high pressures and blowers at
unsteady heat transfer and, as warming or cooling
relatively low pressures. The lower density and
occurs, the thermal capacity (i.e., the size and specific
viscosity of gases results in higher operating speeds
heat), of the system becomes important. Analysis of
and, to minimize leakage, smaller clearance between unsteady heat transfer is complex.
moving parts.

Heat Transfer through a Wall

HEAT TRANSFER
Heat transfer by conduction through walls follows the
Heat transfer in process vessels has been described pre- basic relation given by Fourier’s equation [Eq. (21)],
viously in this encyclopedia.[1] The following is a more which states that the rate of heat flow, Q, is
complete review of heat transfer as a unit operation in proportional to the temperature gradient, dT/dx, and
 3870 Unit Processes in Pharmacy: Fundamentals

to the area normal to the heat flow, A Conversely, the heat flow promoted by a given tem-
perature difference is reduced if the thermal resistance
dT is increased. This is the principle of insulation by lag-
Q ¼
kA ð21Þ
dx ging, and it is illustrated by a composite wall, as shown
in Fig. 4B. If steady-state heat transfer exists, the rate of
As the distance x increases, the temperature T heat transfer is the same for both materials. Therefore,
decreases. Hence, measuring in the x direction, the
temperature gradient, dT/dx, is algebraically negative.
k1 AðT1
T2 Þ k2 AðT2
T3 Þ
The proportionality constant k is the thermal conduc- Q ¼ ¼
tivity. Its numerical value depends on the material of x1 x2
which the body is made and on its temperature.
Metals exhibit high conductivity, although values The major temperature drop occurs across the dis-
vary widely. Nonmetallic solids normally have lower tance x2, indicating that this material provides the
conductivities than do metals. For the porous materi- major thermal resistance. (In the case of heavily lagged,
als of this group, the overall conductivity lies between thin metal walls, the temperature drop and thermal
that of the homogeneous solid and the air that perme- resistance of the metal are so small that they can be
ates the structure. Low resultant values lead to wide ignored). Rearrangement of this equation and the elim-
use as heat insulators. Carbon is an exception among ination of the junction temperature gives Eq. (24).
nonmetals. Its relatively high conductivity and chemi-
cal inertness permit its wide use in heat exchangers. ðT1
T2 Þ
Q ¼ A ð24Þ
Steady nondirectional heat transfer through a plane ðx1 =k1 þ x2 =k2 Þ
wall of thickness x and area A is represented in Fig. 4A.
Assuming that thermal conductivity does not change
Equations of this form can be applied to any number
with temperature, the temperature gradient is linear
of layers.
and equal to (T1
T2)/x, where T1 is the temperature
of the hot face and T2 the temperature of the cool face.
Eq. (21) then becomes Eq. (22)
Heat Transfer in Pipes and Tubes
T1
T2
Q ¼ kA ð22Þ
x Pipes and tubes are common as barriers over which
This may be rearranged to Eq. (23) heat exchange takes place. Conduction is complicated
in this case by the changes in the area over which heat
T1
T2 is transferred. If Eq. (21) is to be retained, some value
Q ¼ A ð23Þ
x=k of A must be derived from the length of the pipe, l,
and the internal and external radii, r1 and r2, respect-
Unit–Validation

where x/k is the thermal resistance. Thus, for a given

ively. If the pipe is thin-walled and the ratio r2/r1 is less
heat flow, a large temperature drop must be created
than approximately 1.5, the heat transfer area can be
if the wall or layer has a high thermal resistance.
based on an arithmetic mean of the two radii, and
Eq. (21) becomes Eq. (25).
A B
r2 þ r1 T 1
T 2
Q ¼ k2 p l ð25Þ
2 r2
r1
T1
T2 This equation is inaccurate for thick-walled pipes,
T1
where the heat transfer area must be based on a logar-
ithmic mean radius rm. Heat transfer is then expressed
by Eq. (26)
T2
T1
T2
T3 Q ¼ k2 prml ð26Þ
Q r2
r1
Q

x x1 where
x2
r2
r1
Fig. 4 Conduction of heat through (A) a plane wall and rm
(B) a composite wall.
logrer2 1
Unit Processes in Pharmacy: Fundamentals 3871

Heat Exchange between a Fluid at a boundary. The approximate evaluation of these

and a Solid Boundary coefficients is discussed later.
The film coefficient h2 may be used to characterize
Conduction and convection contribute to the transfer heat transfer from the barrier to the colder fluid.
of heat from a fluid to a boundary. The distribution
of temperatures at a plane barrier separating two fluids Q Qxw
may be considered. If the fluids are turbulent motion, T1
T1 wall ¼ T1 wall
T2 wall ¼
h1 A hw A
temperature gradients are confined to a relatively
narrow region adjacent to the wall. Outside this region,
where kw is the thermal conductivity of the wall, and
turbulent mixing, the mechanism of which has been
explained earlier, is very effective in the transfer of
heat. Temperature gradients are quickly destroyed Q
T2 wall
T2 ¼
and equalization at values T1 and T2 occurs. Within h2 A
the region, there exists a laminar sublayer across which
heat is transferred by conduction only. The tempera- Addition and rearrangement of these equations gives
ture gradients produced by a given heat flow are corre- Eq. (28).
spondingly high. Outside the laminar layer, eddies
contribute to the transfer of heat by moving fluid from A
the turbulent bulk to the edge of the sublayer, where Q ¼ ðT1
T2 Þ ð28Þ
ð1=h1 þ xw =kw þ 1=h2 Þ
heat can be lost or gained, and by corresponding
movements in the opposite direction. The temperature
The quantity
gradients in this region, where both convection and
conduction contribute to heat transfer, are smaller 1
than in the sublayer.
ð1=h1 þ xw =kw þ 1=h2 Þ
The major resistance to the flow of heat resides in
the laminar sublayer. Its thickness, therefore, is of criti-
cal importance in determining the rate of heat transfer is called the overall heat-transfer coefficient U. A
from the fluid to the boundary. It depends on the general expression of the rate of heat transfer then
physical properties of the fluid, the flow conditions, becomes Eq. (29).
and the nature of the surface. Increase in flow velocity,
for example, decreases the thickness of the layer and, Q ¼ UADT ð29Þ
therefore, its resistance to heat flow.
To evaluate the rate of heat transfer at a boundary, or
a film, transmitting heat only by conduction is postu- 3  
ql l DTagr2 Cp Z q

Unit–Validation
lated. This fictitious film presents the same resistance ¼ constant ð30Þ
to heat transfer as the complex turbulent and laminar DTk Z2 k
regions near the wall. If on the hot side of the wall
the fictitious layer had a thickness x1, the equation of Heat transfer by free convection can thus be pre-
heat transfer to the wall would be sented as a relation between three dimensionless
groups: CpZ/k is known as the Prandtl number, the
T1
T1 wall combination l3DTagr2/Z2 as the Grashof number,
Q ¼ kA
x1 and ql/DT, the Nusselt number may also be written
hl/k.
where k is the thermal conductivity of the fluid. A simi- The specific relation in which these groups stand is
lar equation applies to heat transfer at the cold side of established for a particular system experimentally.
the wall. The thickness of the layer is determined by the The fluid properties, Cp, k, Z, and r are themselves
same factors that control the extent of the laminar sub- temperature dependent. In establishing a correlation,
layer. In general the layer thickness is not known and the temperature at which these properties are to be
the above equation is rewritten as Eq. (27). measured must be chosen. This is usually the tempera-
ture of the main body of the fluid or the mean of this
Q ¼ h1 AðT1
T1 wall Þ ð27Þ temperature and the temperature of the surface.
The exponents r and q are usually found experimen-
where h1 is the heat-transfer coefficient for the film tally to be equal to 0.25 in streamline flow and 0.33 in
under discussion. It corresponds to the ratio k/x1 turbulent flow. The constant varies with the configur-
and is the Btu/ft2  h   F. It is a convenient numerical ation. As an example, the heat transfer to gases and
expression of heat flow by conduction and convection liquids from a large horizontal pipe by free convection
 3872 Unit Processes in Pharmacy: Fundamentals

is described by Eq. (31) is derived only from heating effects. In other systems,
the boiling liquid may be driven through or over
 3  
qd d DTagr2 Cp Z 0:25 heaters, a process referred to as boiling with forced
¼ 0:47 ð31Þ circulation.
kDT Z2 k

The linear dimension in this correlation is the pipe Pool boiling

diameter d. The fluid properties are to be measured
at the mean of the wall and bulk fluid temperatures. If a horizontal heating surface is in contact with a
In forced convection, the fluid is moved over boiling liquid, a sequence of events occurs as the tem-
the surface by a pump or blower neglecting natural perature difference between the surface and the liquid
convection are usually neglected. The study of forced increases.
convection is of great practical importance and vast When DT is small, the degree of superheating of the
amount of data have been amassed for streamline liquid layers adjacent to the surface is low and bubble
and turbulent flow in pipes, across and parallel to formation (growth and disengagement), if present, is
tubes, across plane surfaces, and in other important slow. Liquid disturbance is small and heat transfer
configurations such as jackets and coils. can be estimated from expressions for natural convec-
In forced convection, the heat transferred per unit tion given, for example, in Eq. (29).
area per unit time q, is determined by a linear dimen- Vapor formation becomes more vigorous and bub-
sion which characterizes the surface l, the temperature ble chains rise from points that progressively increase
difference between the surface and the fluid, DT, the in number and finally merge. This movement increases
viscosity Z, the density r, and the velocity u, of the liquid circulation. This phase is called nucleate boiling
fluid, its conductivity k, and its specific heat Cp. and is the practically important regime. A peak flux
Dimensional analysis yields Eq. (32) occurs and a maximum heat-transfer coefficient is
obtained. At this point, DT is known as the critical
   
qd Cp Z x ulr y temperature drop. For water, the value lies between
¼ constant ð32Þ 40 and 50 F (4–10 C). The critical temperature drop
kDT k Z
for organic liquids is somewhat higher. Beyond the
where ql/kT is the Nusselt number, Nu, CpZ/k is the critical temperature, vapor formation is so rapid that
Prandtl number, Pr, and ulr/Z is the Reynolds num- escape is inadequate and a progressively larger fraction
ber, Re, a parameter discussed previously. The values of the heating surface becomes covered with a vapor
of the indices, x and y and of the constant are estab- film. This represents a transition from nucleate boiling
lished for a particular system experimentally. In the to film boiling. When this transition is complete, the
case of turbulent flow in pipes, the correlation for vapor entirely covers the surface, film boiling is fully
fluids of low viscosity is given by Eq. (33). established, and the heat flux rises again.
Unit–Validation

The low heat-transfer coefficient renders film boil-

Nu ¼ 0:023 Pr x Re0:8 ð33Þ ing undesirable and equipment is designed for and
operated at temperature differences which are less than
where x has the value 0.4 for heating and 0.3 for cool- the critical temperature drop. If a constant tempera-
ing. The linear dimension used to calculate Re or Nu is ture heat source such as steam or hot liquid is
the pipe diameter, and the physical properties of the employed, exceeding the critical temperature drop
fluid are to be measured at the bulk fluid temperature. results simply in a drop in heat flux and process
This relation shows that in a given system, the film efficiency. If, however, a constant heat-input source is
coefficient varies as the fluid velocity0.8. If the flow used, as in electrical heating, decreasing heat flux as
velocity is doubled, the film coefficient increases by a the transition region is entered causes a sudden and
factor of 1.7. possibly damaging increase in the temperature of the
heating element; such damage is known as boiling
burn-out.
Heat Transfer to Boiling Liquids Boiling heat-transfer coefficients depend upon both
the physical character of the liquid and the nature of
Heat transfer to boiling liquids occurs in a number of the heating surface. Through the agencies of wetting,
operations, for example, distillation and evaporation. roughness, and contamination, the latter greatly influ-
Heat is transferred by both conduction and convection ences the formation, growth, and disengagement of
in a process further complicated by the phase change bubbles in the nucleate boiling regime. There is, at
that occurs at the heating boundary. When boiling is present, no reliable method of estimating the boiling
induced by a heater in contact with a pool of liquid, coefficients of heat transfer from the physical proper-
the process is known as pool boiling. Liquid movement ties of the system.
Unit Processes in Pharmacy: Fundamentals 3873

Boiling inside a vertical tube high velocity vapor (Fig. 5F). In long tubes, the main
heat transfer takes place in this region by either forced
Heat transfer to liquids boiling in vertical tubes is com- convection or nucleate boiling. At low temperature
mon in evaporators. If a long tube of suitable diameter differences between wall and film, heat transfer occurs
in which liquid lies at a low level is heated, the pattern quietly as in forced convection. This is the normal
of boiling is established Fig. 5. At low levels, boiling regime in a climbing film evaporator and heat flux
may be suppressed by the imposed head (Fig. 5A). can be calculated from correlations of the type given
Higher in the tube, bubbles are produced, which rise in Eq. (40). At high temperatures differences, nucleate
and coalesce (Fig. 5B). Slug formation due to bubble boiling takes place in the film and the vigorous move-
coagulation occurs (Figs. 5C and D). The slugs finally ment leads to an increase in heat transfer coefficient.
break down (Fig. 5E). Escape is hindered and both
liquid and vapor move upward at an increasing speed. Boiling with forced circulation
Draining leads to separation of the phases, giving an
annular film of liquid dragged upward by a core of In many systems, movements other than those caused
by boiling are imposed. For example, boiling in agi-
tated vessels is common in many batch processes.
The boiling heat-transfer coefficients depend upon
A B C
the properties of the liquid, the nature of the surface,
and the agitation. The coefficients obtained are slightly
higher than those of pool boiling. Inside the tubes, the
pattern of forced circulation boiling is similar to that
described in the previous section. Coefficients, how-
ever, are higher because the velocities attained are
higher.

Heat Transfer from Condensing Vapors

When a saturated vapor is brought into contact with a

cool surface, heat is transferred to the surface and a
liquid condenses. The vapor may consists of a single
substance or a mixture, some components of which
may be non-condensable.
The process is described by the following sequence.
The vapor diffuses to the boundary where actual

Unit–Validation
D E F
condensation takes place. In most cases, the conden-
sate forms a continuous layer over the cooling surface,
draining under the influence of gravity. This is known
as film condensation. The latent heat liberated is trans-
ferred through the film to the surface by conduction.
Although this film offers considerable resistance to
heat flow, film coefficients are usually high.

Dropwise condensation

Under some surface conditions, the condensate does

not form a continuous film. Droplets are formed which
grow, coalesce, and then run from the surface. As a
fraction of the surface is always directly exposed to
the vapor, film resistance is absent, and heat-transfer
coefficients, which may be ten times those of film
Fig. 5 Boiling in a narrow vertical tube. (A) Boiling condensation, are obtained. This process is known as
suppressed by head, natural convection is shown; (B) bubble dropwise condensation. Although highly desirable, its
formation; (C) slug formation due to bubble coagulation; (D) occurrence, which depends upon the wettability of
fully developed slug flow; (E) breakdown of slugs at high the surface, is not predictable and cannot be used as
vapor rates; (F) annular-flow-climbing film. a basis for design.
 3874 Unit Processes in Pharmacy: Fundamentals

Condensation of a pure vapor that of a body which absorbs all and reflects none of
the incident radiation, is called a black body.
For film condensation, a theoretical analysis of the
laminar flow of a liquid film down an inclined surface
Exchange of radiation
and the progressive increase in thickness due to
condensation yields Eq. (34) for the mean-transfer
The exchange of radiation is based upon two laws. The
coefficient hm
first, known as Kirchoff’s law, states that the ratio of
 0:25 the emissive power to the absorptivity is same for all
r2 k3 lg bodies in thermal equilibrium. The emissive power of
hm ¼ constant ð34Þ
DTZx a body, E, is the radiant energy emitted from unit area
in unit time. A body of area A1 and emissivity E1,
where l is the latent heat of vaporization; r, k, and Z therefore, emits energy at a rate E1A1. If the radiation
are the density, thermal conductivity, and viscosity of falling on unit area of the body is Eb, the rate of energy
the liquid, respectively; DT is the difference in tem- absorption is Eba1A1, where a1 is the absorptivity. At
perature between the surface and vapor. Experimen- thermal equilibrium, Eba1A1. For another body in the
tally determined coefficients confirm the validity of same environment, Eba1A2 ¼ E2A2, leading to Eq. (36)
Eq. (34). In practice, however, the coefficients are
somewhat higher due to disturbance of the film arising
E1 E2
from a number of actors. As the condensation rate Eb ¼ ¼ ð36Þ
rises, the thickness of the condensate layer increases a1 a2
and the film coefficient falls. However, a point may
be reached in long vertical tubes at which flow in the For a black body, a ¼ 1. The emissive power is
layer becomes turbulent. Under these conditions, the therefore Eb. The black body is a perfect radiator
coefficient rises again and Eq. (34) is not valid. Coeffi- and is used as the comparative standard for other sur-
cients may also be increased if high vapor velocities faces. The emissivity e of a surface is defined as the
induce ripples in the film. ratio of the emissive power E of the surface to
the emissive power of a black body at the same tem-
perature Eb, as shown by Eq. (37).
Condensation of mixed vapors

If a mixture of condensable and noncondensable gases E

e ¼ ð37Þ
is cooled below its dew point at a surface, the former Eb
condenses, leaving the adjacent layers richer in the
latter, thus creating an added thermal resistance. The Emissivity is numerically equal to absorptivity. As
condensable fraction must diffuse through this layer
Unit–Validation

emissive power varies with wavelength, the ratio

to reach the film of condensate and heat-transfer should be quoted at a particular wavelength for many
coefficients are normally very much lower than the materials. However, the emissive power is a constant
corresponding value for the pure vapor. For example, fraction of the black body radiation, that is, the emiss-
the presence of 0.5% of air has been found to reduce ivity is constant. These materials are known as gray
the heat transfer by condensation of steam by as much bodies.
as 50%. The second fundamental law of radiation, known as
the Stefan–Boltzmann law, states that the rate of
energy emission from a black body is proportional to
Heat Transfer by Radiation the fourth power of the absolute temperature T, as
shown by Eq. (38)
Of the radiation that falls on a body, a fraction a is
absorbed, a fraction r is reflected, and a fraction t is
E ¼ sT 4 ð38Þ
transmitted. These fractions are called absorptivity,
reflectivity, and transitivity, respectively. Most indus-
trial solids are opaque such that the transmissivity is where E is the total emissive power and s is the Stefan–
zero and Eq. (35) holds. Boltzmann constant. It is sufficiently accurate to say
that the heat emitted in unit time Q from a black body
a þ r ¼ 1 ð35Þ of area A is given by
Reflectivity, and therefore, absorptivity depend
greatly on the nature of the surface. The limiting case, Q ¼ sAT 4
Unit Processes in Pharmacy: Fundamentals 3875

and for a body that is not perfectly black by Eq. (39) one region to another causes mixing of the components
of the gas. This is called eddy diffusion. Eddy diffusion
Q ¼ seAT 4 ð39Þ is a more rapid process and although molecular dif-
fusion is still present, its contribution to the overall
movement of material is small. In still air, eddy dif-
where e is the emissivity. fusion is virtually absent and evaporation occurs by
The net energy gained or lost by a body can be esti- molecular diffusion.
mated by these laws. The simplest case is that of a gray
body in black surroundings. These conditions, in which
none of the energy emitted by the body is reflected
back, are approximately those of a body radiating to Molecular Diffusion of Gases
atmosphere. If the absolute temperature of the body
is T1, the rate of heat loss is seAT14 [Eq. (40)], where Transport of material in stagnant fluids or across
A is the area of the body and e its emissivity. Surround- streamlines of a fluid in laminar flow occurs by molecu-
ings at a temperature T2 emit radiation proportional to lar diffusion. Two adjacent compartments, separated
sT24, and a fraction, determined by area and absorp- by a partition containing pure gases A or B may be
tivity a, is absorbed by the body; this heat is saAT24, envisaged. Random movement of all molecules occurs
and as absorptivity and emissivity are equal, Eq. (40) so that after a period tome, molecules are found quite
is valid. remote from their original positions. If the partition is
removed, some molecules of A move toward the region
Net heat-transfer rate ¼ seAðT14
T24 Þ ð40Þ occupied by B, their number depending on the number
of molecules at the point considered. Concurrently,
If part of the energy emitted by a surface is reflected molecules of B diffuse toward regimes formerly occu-
back by another surface, the calculation of radiation pied by pure A.
exchange is more complex. Equations for various sur- Ultimately, complete mixing occurs. Before this
face configurations take the following general form: point in time, a gradual variation in the concentration
of A occurs along an axis, designated x, which joins
Q ¼ F1 F2 sAðTA4
TB4 Þ the original compartments. This variation, expressed
mathematically, is
dCA/dx, where CA is the concen-
where F1 and F2 are factors determined by the con- tration of A. The negative sign arises because the con-
figuration and emissivity of surfaces at temperatures centration of A decreases as the distance x increases.
TA and TB. Similarly, the variation in the concentration of gas B
is
dCB/dx. The rate of diffusion of A, NA, depends
on the concentration gradient and the average velocity
with which the molecules of A move in the x direction.

Unit–Validation
MASS TRANSFER
This relationship is expressed by Fick’s law, given by
Eq. (41)
Mass transfer in unit operations has been described in
detail previously in this encyclopedia.[2] The following
is a brief review to complete the overview of unit pro- d=CA
cesses in pharmacy. NA ¼
DAB ð41Þ
dx
In mass transfer operations, two immiscible phases
are normally present, one or both of which are fluid.
In general, these phases are in relative movement and where D is the diffusivity of A in B, a property
the rate at which a component is transferred from proportional to the average molecular velocity and,
one phase to the other is greatly influenced by the bulk therefore, dependent on the temperature and pressure
movement imposed upon the fluids. In most drying of the gases. The rate of diffusion NA is usually
processes, for example, water vapor diffuses from a expressed as the number of moles diffusing across unit
saturated layer in contact with the drying surface into area in unit time. As with the basic equations of heat
a turbulent air stream. The boundary layer, as transfer, Eq. (41) indicates that the rate of a process
described earlier, consists of a sublayer in which flow is directly proportional to a driving force, which, in
is laminar and an outer region in which flow is tur- this context, is a concentration gradient.
bulent. The mechanism of diffusion differs in these This basic equation can be applied to a number
regimes. In the laminar layer, movement of water of situations. Restricting discussion exclusively to
vapor molecules across streamlines can occur only by steady-state conditions, in which neither dCA/dx or
molecular diffusion. In the turbulent region, the move- dCB/dx change with time, equimolecular counter
ment of relatively large units of gas, called eddies, from diffusion is considered first.
 3876 Unit Processes in Pharmacy: Fundamentals

Equimolecular counterdiffusion movement must be balanced by bulk flow away from

the surface. The total flow of A is, therefore, the diffu-
If no bulk flow occurs in an element of length dx, the sional flow of A plus the transfer of A associated with
rates of diffusion of the two gases A and B must be this bulk movement.
equal and opposite, that is, NA ¼
NB.
The partial pressure of A changes by dPA over the
Molecular Diffusion in Liquids
distance dx. Similarly, the partial pressure of B changes
dPB. As there is no difference in total pressure across
Equations describing molecular diffusion in liquids are
the element (no bulk flow), dPA/dx must equal
similar to those applied to gases. The rate of diffusion

dPB/dx. for an ideal gas, the partial pressure is
of material A in a liquid is given by Eq. (40).
related to the molar concentration by the relation
dCA
PA V ¼ nA RT NA ¼ D
dx
where nA is the number of moles of gas A in a volume Fick’s law for steady-state equimolal counterdiffusion
V. As the molar concentration CA is equal to nA/V, is then expression by Eq. (44)
therefore
C A 2
CA 1
PA ¼ CA RT NA ¼
D ð44Þ
x2
x1

Consequently, for gas A, Eq. (40) can be written as in where CA2 and CA1 are the molar concentrations at
Eq. (42), points x2 and x1 respectively.
Equations for diffusion through a layer of stagnant
DAB dPA liquid can also be developed. The applicability of these
NA ¼
ð42Þ
RT dx equations is, however, limited because diffusivity in a
liquid varies with concentration. In addition, unless
where DAB is the diffusivity of A in B. Similarly, the solutions are very dilute, the total molar concen-
DBA dPB DAB dPA tration varies from point to point. These complications
NB ¼
¼ do not arise with diffusion in gases.
RT dx RT dx
Diffusivities in liquids are very much lower than dif-
It, therefore, allows that DAB ¼ DBA ¼ D. If the partial fusivities in gases, commonly by a factor of 104.
pressure of A at x1 is PA1 and at x2 is PA2, integration of
Eq. (42) gives Eq. (43). Mass Transfer in Turbulent and Laminar Flow
 
Unit–Validation

D P A2
P A1
NA ¼
ð43Þ As already explained, movement of molecules across
RT x2
x1 the streamlines of a fluid in laminar flow can occur
only by molecular diffusion. If the concentration of a
A similar equation may be derived for the counter-
component A varies in a direction normal to the
diffusion of gas B.
streamlines, the molar rate of diffusion is given by
Eq. (44).
Diffusion through a stationary, non-diffusing gas When a fluid flows over a surface, the surface
retards the adjacent fluid region, forming a boundary
An important practical case arises when a gas A dif- layer, If the flow throughout the fluid is laminar, the
fuses through a gas B, there being no overall transport equation for molecular diffusion may be used to evalu-
of gas B. It arises, for example, when a vapor formed at ate the mass transferred across the boundary layer. In
a drying surface diffuses into a surrounding gas. At the most important cases, however, the flow in the bulk of
liquid surface, the partial pressure of A is dictated by the fluid is turbulent. The boundary layer is then con-
the temperature. For water, it would be 12.8 mm Hg sidered to consist of three distinct flow regimes. In the
(1.7 kPa) at 15 C. Some distance away the partial region of the boundary layer most distant from the sur-
pressure is lower, and the concentration gradient face, flow is turbulent, and mass transfer is the result of
causes diffusion of A away from the surface. Similarly, the interchange of large portions of the fluid. Mass
a concentration gradient for B must exist, the concen- interchange is rapid and concentration gradients are
tration being lowest at the surface. Diffusion of this low. As the surface is approached, a transition from
component takes place toward the surface. There is, turbulent to laminar flow occurs in the transition or
however, no overall transport B so that diffusional buffer region. In this region, mass transfer by eddy
Unit Processes in Pharmacy: Fundamentals 3877

diffusion and molecular diffusion are of comparable However, x0 is not known, and Eq. (45) may be written
magnitude. In a fluid layer at the surface, a fraction as
of a millimeter thick, laminar flow conditions persist.
This laminar sublayer, in which transfer occurs by mol- kg
ecular diffusion only, offers the main resistance to NA ¼
ðPAi
PAg Þ ð45Þ
RT
mass transfer as shown in Fig. 6A. As flow becomes
more turbulent, the thickness of the laminar sublayer where kg, is a mass transfer coefficient, the units of
and its resistance to mass transfer decrease. which are cm sec
1. As CA ¼ PA/RT, it can also be
An approach to the evaluation of the rate of written as
mass transfer under these conditions lies in the postu-
lation of a film, the thickness of which offers the same NA ¼ kg ðCAi
CAg Þ
resistance to mass transfer as the combined laminar,
buffer, and turbulent regions. The analogy with where CAi and CAg are the gas concentrations at either
heat transfer by conduction and convection is exact side of the film. Similar equations describe the dif-
and quantitative relations between heat and mass fusion of B in the opposite direction.
transfer can be developed for some situations. This, Diffusion across a liquid film is described by Eq. (46)
however, is not attempted here. The postulate of an
effective film is explained by Fig. 6B. As a gas flows NA ¼ k1 ðCAi
CA1 Þ ð46Þ
over a surface, equimolecular counterdiffusion of com-
ponents A and B occurs, A away from the surface and where CAi is the concentration of component A at the
B toward the surface. The variation in partial pressure interface and CA1 is its concentration in the bulk of the
of A with distance from the surface is shown in Fig. 6A. phase.
At the surface, the value of PAi, a linear fall to PAb In all cases, the mass-transfer coefficient depends
occurs over the laminar sublayer. Beyond this, the upon the diffusivity of the transferred material and
partial pressure falls less steeply to the value PA at the thickness of the effective film. The latter is largely
the edge of the boundary layer. A value slightly higher determined by the Reynolds number of the moving
than this is PAg, the average partial pressure of A fluid, that is, its average velocity, density, and viscosity,
in the entire system. In general, the gas content of and some linear dimension of the system. Dimensional
the laminar layer is so small that PA and PAg are analysis gives the following relation:
virtually equal. If the molecular diffusion were solely
responsible for diffusion, the partial pressure PAg  r
kd Z
would be reached at some fictitious distance x0 from ¼ constðReÞ q

the surface, over which the concentration gradient D rD

(PAi
PAg)/x0 exists. The molar rate of mass transfer
where Re is the Reynolds number, k the mass transfer

Unit–Validation
would then be
coefficient, D the diffusivity, and d a dimension charac-
D PAi
PAg terizing the geometry of the system.
NA ¼ 
RT x This relation is analogous to the expression for the
heat transfer by forced convection given earlier. The
dimensionless group kd/D corresponds to the Nusselt
group in heat transfer. The parameter Z/rD is known
A B as the Schmidt number and is the mass-transfer
Gas Liquid
counterpart of the Prandtl number. For example, the
evaporation of a thin liquid film at the wall of a pipe
Laminar sublayer

Interface into a turbulent gas is described by the equation

PAi
Partial pressure of A

CAi
Concentration

PAg kd
¼ 0:023 Re0:8 Sc0:33
D
PAi
PAb
CAL
where Sc is the Schmidt number. Although this equa-
tion expresses experimental data, comparison with
PA Eq. (41) again demonstrates the fundamental relation
Direction of diffusion
x'
of heat and mass transfer.
PAg
Distance from surface Distance
Similar relations have been developed empirically
for other situations. The flow of gases normal to
Fig. 6 Mass transfer at (A) a boundary and (B) an interface. and parallel to liquid surfaces can be applied to drying
 3878 Unit Processes in Pharmacy: Fundamentals

processes, and the agitation of solids in liquids can pro- concentration. The rate of solution is determined by
vide information for crystallization or dissolution. the difference between the interfacial concentration
and the concentration in the bulk solution, and the
mass-transfer coefficient.
Interfacial Mass Transfer

On a macroscopic scale, the interface can be regarded

as a discrete boundary. On the molecular scale, how- REFERENCES
ever, the change from one place to another takes place
over several molecular diameters. Due to movement of 1. Markovitz, R.E. Heat transfer in process vessels. In Encyclo-
molecules, this region is in a state of violent change, the pedia of Pharmaceutical Technology, 2nd Ed.; Swarbrick,
J., Boylan, J.C., Eds.; Marcel Dekker, Inc.: New York, 2001.
whole surface layer changing many times a second. 2. Graham Nairn, J. Mass transfer in unit operations. In
Transfer of molecules at the actual interface is, there- Encyclopedia of Pharmaceutical Technology, 2nd Ed.;
fore, virtually instantaneous and the two phases are, Swarbrick, J., Boylan, J.C., Eds.; Marcel Dekker, Inc.: New
York, 2001.
at this point in equilibrium.
As the interface offers no resistance, mass transfer
between phases can be regarded as the transfer of a
component from one bulk phase to another through BIBLIOGRAPHY
two films in contact, each characterized by a mass-
transfer coefficient. This is the two-film theory and Beard, J. Dynamics of Fluids in Porous Media; Dover Publica-
tions, Inc.: Mineola, NY, 1972.
the simplest of the theories of interfacial mass transfer. Bird, R.B.; Stewart, W.E.; Lightfoot, E.N. Transport Phenom-
For the transfer of a component from a gas to a liquid, ena; John Wiley & Sons: New York, 1960.
the theory is described in Fig. 6B. Across the gas film, Chhabra, R.P. Bubbles, Drops and Particles in Non-Newtonian
Fluids; CRC Press: Boca Raton, FL, 1993.
the concentration, expressed as partial pressure, falls Cheremisinoff, N.P. Practical Fluid Mechanics for Engineers
from a bulk concentration PAg to an interfacial con- and Scientists; Technomic Publishing Co., Inc.: Lancaster,
centration PAi. In the liquid, the concentration falls PA, 1990.
Cho, Y.I.; Hartnett, J.P. Non-newtonian fluids in circular pipe
from an interfacial value CAi to bulk value CA1. flow. Adv. Heat Transfer 1982, 15, 59–141.
At the interface, equilibrium conditions exist. The Coulson, J.M.; Richardson, J.R. Chemical Engineering, 3rd Ed.;
break in the curve is due to the different affinity of Pergamon Press: New York, 1977; 1.
Crank, J. Ed.; The Mathematics of Diffusion, 2nd Ed.;
component A for the two phases and the different units Crank, J., Ed.; Oxford University Clarendon Press: Oxford,
expressing concentration. The bulk phases are not, 1975, reprinted 1992.
of course, at equilibrium and it is the degree of dis- Cussler, E.L. Diffusion Mass Transfer in Fluid Systems; Cam-
bridge University Press: New York, 1984.
placement from equilibrium conditions that provides Fayed, M.E.; Otten, L. Handbook of Powder Science and Tech-
the driving force for mass transfer. If these conditions nology; Van Nostrand Reinhold, Co.: New York, 1984.
are known, an overall mass-transfer coefficient can be Hartnett, J.P.; Kostic, M. Heat transfer to newtonian and non-
Unit–Validation

Newtonian fluids in rectangular ducts. Adv. Heat Transfer

calculated and used to estimate the mass-transfer rate. 1989, 19, 24–356.
Transfer of a component from one mixed phase to McCabe, W.L.; Smith, J.C.; Harriott, P. Unit Operations of
another, as described above, occurs in several pro- Chemical Engineering, 5th Ed.; McGraw Hill, Inc.: New
York, 1993.
cesses. Liquid–liquid extraction, leaching, gas absorp- Meyer, R.E. Introduction to Mathematical Fluids Dynamics;
tion, and distillation are examples. In other processes Dover Publications, Inc.: Mineola, NY, 1982.
such as drying, crystallization, and dissolution, one Neumann, B.S. The flow properties. In Advances in Pharmaceu-
tical Sciences; Bean, H.S., Beckett, A.H., Carless, J.E., Eds.;
phase may consist of only one component. Concen- Academic Press: New York, 1967; 2, 181–221.
tration gradients are set up in one phase only, with Orr, C. Particulate Technology; Macmillan: New York, 1966.
the concentration at the interface given by the relevant Ozisik, M.N. Boundary Value Problems of Heat Conduction;
Dover Publications, Inc.: Mineola, NY, 1968.
equilibrium conditions. In drying, for example, a layer Prandtl, L.; Tietjens, O.G. Fundamentals of Hydro- an Aero-
of air in equilibrium (i.e., saturated) with the liquid is mechanics; Dover Publications, Inc.: Mineola, NY, 1957.
postulated at the liquid surface and mass transfer to Perry, R.H.; Chilton, C.H. Chemical Engineer’s Handbook,
5th Ed.; McGraw Hill: New York, 1973.
a turbulent air stream is described by Eq. (44), Taylor, R.; Krishna, R. Multicomponent Mass Transfer; John
the interfacial concentration being the saturation Wiley & Sons: New York, 1993.
Unit Processes in Pharmacy: Operations
Anthony J. Hickey
Department of Pharmaceutics, The University of North Carolina at Chapel Hill,
Chapel Hill, North Carolina, U.S.A.

David Ganderton
Devon, U.K.

INTRODUCTION heating fluid, the temperature difference across the

wall of an evaporator is determined by the boiling tem-
The fundamentals of unit processes have been perature, a variable controlled by the external pressure
described elsewhere.[1,2] The following is a summary and the concentration of the solute in the solution. The
of the major operations. Although it is broad in scope, boiling temperature and the solute concentration both
the abbreviated form of this article has resulted in influence the viscosity of the solution, a factor which
selected omissions. References are given, wherever greatly affects the heat transfer coefficient. The boiling
possible, to make up for this deficiency. temperature also determines the solubility of dissolved
constituents and the degree of concentration which can
be carried out without separation of solids.
EVAPORATION AND DISTILLATION
The Effect of Heat on the Active Constituents of a
Evaporation Solution. The thermal stability of components of a
solution may determine the type of evaporator to be
The term evaporation, in the pharmaceutical industry, used and the conditions of its operation. If a simple
is primarily associated with the removal of water and solution contains a hydrolyzable material and the rate
other solvents by boiling in batch processes. of its degradation during evaporation depends on its
concentration at any time, an exponential relation
Heat transfer to boiling liquids in an evaporator between the remaining fraction, F, and the time, t,
characteristic of a first-order reaction, is obtained, as
The heat required to boil a liquid in an evaporator is shown in Eq. (2).
usually transferred from a heating fluid, such as steam

Unit–Validation
or hot water, across the wall of a jacket or tube in or F ¼ e
kt ð2Þ
around which the liquid boils. A qualitative discussion
of the methods used to secure high rates of heat flow The dependence of the reaction velocity constant, k,
can be used on Eq. (1), on the absolute temperature, T, is expressed by Eq. (3),

Q ¼ UA DT ð1Þ k ¼ Ae
B=T ð3Þ

where Q is the rate of heat flow, U is the overall heat where A and B are constants characteristic of the reac-
transfer coefficient, A is the area over which heat is tion. Thus, at temperatures T1, T2, and T3, where
transferred, and DT is the difference in temperature T1 > T2 > T3, the relation between the remaining
between the fluids. fraction and the time of heating becomes clear, as
Other factors described by Eq. (1) are the area of the shown in Fig. 1. This indicates the importance of
heat transfer surfaces, which should be as large as pos- the temperature and time of heating. If the latter can
sible, and the temperature difference between the heat- be shortened, the temperature of evaporation can be
ing surface and the boiling liquid. As long as the critical increased greatly without increasing the fraction which
heat flux is not exceeded, the latter also should be large. is degraded. If, therefore, the effect of temperature on
the rate of evaporation is known, it is possible to define
The physical properties of solution and liquids the conditions of time and temperature at which
decomposition is minimum.
A number of physical factors, which are inter-related, In practice, the kinetics of degradation and the
are relevant to the study of evaporation. For a given relation of evaporation rate and temperature are
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-100001926
Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved. 3879
 3880 Unit Processes in Pharmacy: Operations

1.0 escaping to atmosphere or into a vented hood. Larger

pan evaporators are closed, the vapor escaping
through a pipe. Small jacketed pans are efficient and
easy to clean and may be fitted for the vacuum evapora-
Fraction Remaining

tion of thermolabile materials. Their size, however, is

limited because the ratio of heating area to volume
0.5
decreases as the capacity increases. Larger vessels must
employ a heating coil which increases their evaporating
T3 capacity but it also makes cleaning more difficult.

T2
Forced-circulation evaporators

T1 On the smallest scale, forced-circulation evaporators

0.0 are similar to the pan evaporators described priviously,
Time modified only by the inclusion of an agitator. Vigorous
Fig. 1 The effect of time and temperature of degradation.
agitation increases the boiling film coefficient, the
degree of which depends on the type and speed of
the agitator. An agitator should be used for the evap-
usually not known. This is particularly true when the oration of viscous materials to prevent degradation
criteria by which the product is judged are color, taste, of material at the heated surfaces. Some large-scale
or smell. In addition, this analysis neglects the tem- continuous units are similar to the natural circulation
perature variation in the evaporating liquid and the evaporators already described.
degradation in boundary films where temperatures
are higher. Therefore, experiments are often necessary Film evaporators
to determine the suitability of an evaporation process.
In batch processes, the time of exposure to heat is In the short tubes of the calandria, an intimate mixture
well defined. This is also true of continuous processes of vapor and liquid is discharged at the top as shown in
in which the liquid to be evaporated is passed only Fig. 2A. If the length of the tube is greatly increased,
once through the heater. In continuous processes, is progressive phase separation occurs until a high veloc-
where the liquid is recirculated through the heater, ity core of vapor is formed which propels an annular
the average residence time, a, is given by the ratio. film of liquid along the tube. This phenomenon, which
The volumetric discharge is only an indication of is one stage of flow when a liquid and a gas pass in the
the damage that may be caused by prolonged heating. same direction along a tube, is employed in film evap-
If perfect mixing occurs in the evaporator, the fraction, orators. The turbulence of the film gives very high heat
Unit–Validation

f, which is in the unit for time t or less, is given by transfer coefficients, and the bubbles and vapor
Eq. (4). evolved are rapidly swept into the vapor stream.
Although recirculation may be adopted, it is possible
f ¼ 1
e
t=a ð4Þ with the high evaporation is found in long tubes to
sufficiently concentrate the liquid in a single pass.
This relation shows that an evaporator, for Because a very short residence time is obtained, very
example, with an average residence time of 1 h holds
13.5% of active principles for 2 h and about 2% for 4 h.

A B
Vapor Vapor to
Evaporators condenser

It is convenient to classify evaporators into natural cir-

culation evaporators, forced-circulation evaporators,
Feed Steam
and film evaporators. Steam Cyclone
separator
Natural circulation evaporators Concentrate
Condensate
Condensate
Small-scale evaporators consist of a simple pan heated Concentrate
Feed
by a jacket or a coil, or both. Admission of the heating
fluid to the jacket induces the liquid in the vessel to Fig. 2 (A) Evaporator with calandria; (B) climbing film
boil. Very small evaporators may be open, the vapor evaporator.
Unit Processes in Pharmacy: Operations 3881

thermolabile materials may be concentrated at rela- of vapor from the surface is described by Eq. (5),
tively high temperatures. Film evaporators are also
suitable for materials which foam strongly. Various NA ¼ kg RTðPAi
PAg Þ ð5Þ
types have been developed, but all are essentially con-
tinuous in operation and their capacity starts from a where NA is the number of moles evaporating from
few liters per hour upward. unit area in unit time, kg is the mass transfer coefficient
The climbing film evaporator, which is the most across the boundary layer, R is the gas constant, T is
common film evaporator, consists of tubes 15 to 30 feet the absolute temperature, PAi is vapor pressure of the
(4.5–13.6 m) long and 1 to 2 in. (2.5–5 cm) in diameter liquid, and PAg is the partial pressure of the vapor in
mounted in a steam chest (Fig. 2B). The feed liquid the gas stream; kg is proportional to the gas velocity.
enters the bottom of the tubes and flows upward for
a short distance before boiling begins. The length of
this section, which is characterized by low heat transfer Distillation
coefficients, may be minimized by preheating the feed
to its boiling point. The pattern of boiling and phase Distillation is a process in which a liquid mixture is
separation follows and a mixture of liquid and vapor separated into its component parts by vaporization.
emerges from the top of the tube to be separated by The vapor evolved from a boiling liquid mixture is nor-
baffles or by a cyclone separator. Climbing film evap- mally richer in the more volatile components than the
orators are not suitable for the evaporation of viscous liquid with which it is in equilibrium. Distillation rests
liquids. upon this fact. Although multicomponent mixtures are
In the falling film evaporator, the liquid is fed to the most common in distillation processes, an understand-
top of a number of long heated tubes. Because the ing of the operation can be based on the vapor pressure
gravity assists the flow down the tube, this arrange- characteristics of two-component or binary mixtures.
ment is better suited to the evaporation of moderately
viscous liquids. The vapor evolved is usually carried Binary mixtures of immiscible liquids:
downward and the mixture of liquid and vapor steam distillation
emerges from the bottom for separation. Even distri-
bution of liquid must be secured during feeding. If the components of a binary mixture are immiscible,
The tendency to channel in some tubes leads to drying the vapor pressure of the mixture is the sum of the
in others. vapor pressures of the two components, each exerted
The rising-falling film evaporator concentrates a independently and not as a function of their relative
liquid in a climbing film section and leads the emerging concentrations in the liquid. This property is employed
liquid and vapor into a second tube section which in steam distillation, a process particularly applicable
forms a falling film evaporator. Good distribution of to the separation of high boiling substances from

Unit–Validation
liquid is claimed in the falling film section and this type non-volatile impurities. The steam forms a cheap and
is particularly suitable for liquids which greatly inert carrier. The principles of the process also apply
increase in viscosity during evaporation. to other immiscible systems.
In mechanically aided film evaporators, a thin The composition of the distillate is calculated in the
film of material is maintained on the heat transfer sur- following way. For two components, A and B, the total
face irrespective of the viscosity. This is usually vapor pressure, P, is the sum of the vapor pressures of
achieved by means of a rotor, concentric with the tube, the components, PA and PB. Since the partial pressure
which carries blades that either scrape the tube or of a component in a gaseous mixture is proportional to
ride with low clearance in the film. Mechanical agi- its molar concentration, the composition of the vapor
tation permits the evaporation of highly viscous mate- is given by Eq. (6),
rials or those that have a low thermal conductivity.
Because the temperature variations in the film are nA PA
¼ ð6Þ
reduced and residence times are shortened, the vacuum nB PB
evaporation of viscous thermolabile materials becomes
possible. where nA and nB are the number of moles of A and B
in the vapor, respectively. If WA and WB are the weights
Evaporation without boiling of A and B in the vapor, Eq. (7) holds,
WA MB PA
During heating, some evaporation takes place at the ¼ ¼ ð7Þ
MA WB PB
surface of a batch of liquid before boiling begins. Simi-
larly, liquids that are very viscous or excessively froth where MA and MB are the respective molecular weights.
may be concentrated without boiling. The diffusion The distillate obtained from the vapor is WA þ WB,
 3882 Unit Processes in Pharmacy: Operations

and the percentage of A in the distillate is expressed by similarly,

Eq. (8).
xB PB0
yB ¼ ð13Þ
WA PA M A P
 100 ¼  100 ð8Þ
MA þ W B P A M A þ P B WB
If A is the more volatile component, P0A is greater than
The ratio of immiscible organic liquid to water in P. Therefore yA is greater than xA, that is, the vapor is
the distillate is increased if the former has a high richer in the more volatile component than the liquid
molecular weight or a high vapor pressure. Steam with which it is in equilibrium.
distillation under vacuum may be employed when the Systems that form minimum boiling mixtures are
thermal stability of the material prohibits temperatures common. Ethyl alcohol and water provide an example,
of approximately 100 C. the azeotrope containing 4.5% by weight of water. The
boiling point at atmospheric pressure is 78.15 C that is
The Relation of Vapor Pressure and Mixture Compo- 0.25 C lower than the boiling point of pure alcohol.
sition. In a binary mixture of two completely miscible Maximum boiling mixtures are less common. The most
components, the vapor pressure is a function of the familiar example is hydrochloric acid which forms an
mixture composition as well as of the vapor pressures azeotrope boiling at 108.6 C containing 20.2% by
of the two pure components. If the liquids are ideal, weight of hydrochloric acid. Mixtures that form azeo-
the relation of vapor pressure and composition is given tropes cannot be separated into the pure components
by Raoult’s law. At a constant temperature, the partial by normal distillation methods. However, separation
vapor pressure of a constituent of an ideal mixture is into the azeotrope and one pure component is possible.
proportional to its mole fraction in the liquid. Thus,
for a mixture of A and B, the partial vapor pressure Simple or differential distillation
of A is given by Eq. (9),
In simple or differential distillation, the vapor evolved
PA ¼ PA0  xA ð9Þ from the boiling mixture is immediately removed and
condensed. Unless the boiling points of the two pure
where P0A is the vapor pressure of pure A and xA is its components differ widely, a reasonable degree of sep-
mole fraction. Similarly, the partial vapor pressure of B aration is not possible. This method may be used to
is expressed by Eq. (10). remove low boiling solvents from aqueous solutions.

PB ¼ PB0  xB ð10Þ Rectification or fractionation

The total pressure of the system, P, is simply PA þ PB. In simple distillation, vapor enrichment is small. In
Unit–Validation

Very few liquid mixtures rigidly obey Raoult’s law. fractionation, a term synonymous with rectification,
Consequently, the vapor pressure data must be determ- the vapor leaving the boiling liquid is led up a column
ined experimentally. Mixtures that deviate positively to meet a liquid stream or reflux which originates
from this law give a total vapor pressure curve higher in the column as part of the condensate. In a
which lies above the theoretical straight line. Negative series of partial condensations and vaporizations, the
deviations fall below the line. In extreme cases, devia- rising vapor becomes richer in the more volatile
tions are so large that a range of mixtures exhibits a component at the expense of the falling liquid and high
higher or lower vapor pressure than either of the pure degrees of separation become possible. The columns,
components. called fractionating columns, are of two basic types:
Returning to ideal systems, the partial pressure of a packed columns and plate columns.
component in the vapor is proportional to its mole
fraction, as shown in Eq. (11) for component A, Packed Columns. These are used for laboratory and
small-scale industrial distillation and are usually oper-
PA ¼ yA P ð11Þ ated as a batch process. The column consists of a
vertical, hollow, cylindrical shell containing a packing
where PA is the partial pressure of A in the vapor and designed to offer a large interfacial contact area
yA is its mole fraction. Because, PA ¼ P0A  xA, Eqs. (12) between liquid and vapor. The form of the packing
and (13) can be written as: varies but Raschig rings, which consist of small met-
allic or ceramic cylinders, are the most commonly used.
xA PB0 In general, packed columns operate under widely vary-
yA ¼ ð12Þ
P ing conditions without serious loss of efficiency.
Unit Processes in Pharmacy: Operations 3883

Plate Columns. A plate column consists of a series of industrial molecular still shown in Fig. 3, the feed is
plates or trays on which the liquid is retained for some introduced at the bottom of a heated conical rotor and
period during its movement down the column. The ris- flows upward as a thin liquid layer under the action of
ing vapor is bubbled through this liquid, providing centrifugal force. The residue is caught in a gutter at
intimate contact between the phases. Liquid in reflux the top. The vapor is condensed on a concentric, water-
moves downward between plates and is usually carried cooled condenser a short distance away and discharged.
by a downcomer. Contact between the vapor and
liquid takes place in stages.
Plate columns operate efficiently over a limited AIR CONDITIONING AND HUMIDIFICATION
range of conditions. They are mainly used in large-
scale continuous installations in which the conditions General principles of the supply of air in pharmaceuti-
of distillation can be closely maintained. cal processes are similar to conventional air condition-
ing. The control of its quality, however, may be more
Molecular distillation stringent. In areas where sterile materials are made
and handled, for example, the cleaning process must
Molecular distillation is carried out without boiling at remove bacteria. In other situations, it may be neces-
very low pressures of the order 0.001 mm Hg sary to remove water vapor. The flow of powders is a
(0.133 Pa). At these pressures, collision of molecules sensitive function of moisture content, and the equilib-
in the evolving vapor and reflection back to the liquid rium moisture content of a material is determined by
surface is greatly decreased and the mean free path of the humidity. Some tableting processes break down if
the molecules is of the same order as the distance the humidity is too high. In such processes, the scale
between the evaporating surface and a condenser placed of the air conditioning varies. It may be necessary to
a short distance away. It then becomes possible to distil supply a whole room with air of a certain quality.
liquids of very high boiling point although the degree Alternatively, conditioning may be restricted to a small
of separation cannot exceed one theoretical plate. The area surrounding a particular piece of equipment.
process therefore is used primarily to concentrate non-
volatile components in a high boiling medium. The
vitamins in cod liver oil can be concentrated this way. Vapor and Gas Mixtures
For the separation of liquids of comparable volatility,
several separate distillation stages are necessary. The study of the properties of the air–water vapor mix-
Because agitation due to boiling is absent, an alterna- ture is called psychometry, and data are presented in
tive method of maintaining the more volatile component various forms of psychrometric charts presenting vari-
at the evaporating surface must be adopted. In the ous data. In Fig. 4, humidity is plotted as ordinate and

Unit–Validation
Distillate Cooling water
outlet inlet
Outlet
Distilland
gutter Cooling water outlet
Boiler
Heaters

Condensation pump Thermal

insulation
Heat
To fore pump exchanger Radiant heaters
Louvre condenser
Cast aluminum rotor

Cooling water Bearing housing

inlet
Motor
Residue
Boiler feed pump
pump
Residue
outlet Feed pump

Fig. 3 Large-scale molecular still. (From Ref.[3].)

 3884 Unit Processes in Pharmacy: Operations

Equating Eqs. (14) and (15) gives Eq. (16).

100%

80%
0.12

60%

40%
0.11 h
Hi
H a ¼ ðTa
Twb Þ ð16Þ
0.10 rkg l

hum tive
y
idit
0.09

a
Rel
0.08 Both the heat and mass transfer coefficients are
functions of air velocity. However, at air speeds greater
Humidity, H

0.07

%
45°

20
than about 15 ft/s (4.5 m/s), the ratio h : kg is approxi-
0.06 mately constant. The wet-bulb depression is directly
0.05 proportional to the difference between the humidity
%
10 at the surface and the humidity in the bulk of the air.
0.04 35°
In the wet- and dry-bulb hygrometer, the wet-bulb
0.03
25° depression is measured by two thermometers, one of
0.02 15° B which is fitted with a fabric sleeve wetted with water.
A These thermometers are mounted side by side and
0.01 5°
shielded from radiation, an effect neglected in the deri-
0.00
0 10 20 30 40 50 60 70 80 vation above. Air is drawn over the thermometers by
Temperature, °C means of a small fan. The derivation of the humidity
from the wet-bulb depression and a psychrometric
Fig. 4 A psychrometric chart.
chart are discussed later.
Many wet- and dry-bulb hygrometers operate with-
temperatures as abscissa. Percentage of relative out any form of induced air velocity at the wet bulb.
humidity is plotted as curves running across the chart. This may be explained by examining another air–water
The use of this simplified chart is demonstrated later. system. If a limited quantity of air and water is allowed
to equilibrate under conditions in which heat is neither
gained nor lost by the system, the air becomes satu-
Hygrometry, the Measurement of Humidity rated and the latent heat required for evaporation is
drawn from both fluids which cool to the same tem-
The accurate determination of the humidity of air is perature. This temperature is the adiabatic saturation
carried out gravimetrically. The water vapor present temperature, T1. It is a peculiarity of the air–water sys-
in a known volume of air is chemically absorbed with tem that the adiabatic saturation temperature and the
a suitable reagent and weighed. In less laborious meth- wet-bulb temperature are the same. If water at this
ods, the humidity is derived from the dew point or the temperature is recycled in a system through which air
wet-bulb depression of a water–vapor mixture. is passing, the incoming air is cooled till it reaches
Unit–Validation

The dew point is the temperature at which a vapor the adiabatic saturation temperature at which point it
condenses when cooled at constant pressure. If air of is saturated. The temperature of the water, on the other
the condition denoted by point A in Fig. 4 is cooled, hand, remains constant and all the latent heat required
the relative humidity increases until the mixture is fully for evaporation is drawn from the sensible heat of the
saturated. This condition is given by point B; the tem- air. Equilibrium is expressed by Eq. (17),
perature coordinator is the dew point, which can be
measured rapidly by evaporating ether in a silvered
ðTa
T1 ÞS ¼ ðH1
Ha Þl ð17Þ
bulb. The temperature at which dew deposits from
the surrounding air is noted and the humidity is read
directly from a psychrometric chart. where Ta is the temperature of the incoming air and S is
The derivation of the humidity from the wet-bulb its specific heat, Ha and H1 are the humidities of the
depression requires a preliminary study of the transfer incoming air and the saturated air, and l is the latent
of mass and heat at a boundary between air and water. heat of evaporation for water.
The difference between the air temperature and the When both wet- and dry-bulb temperatures have
wet-bulb temperature is the wet-bulb depression. If been found, the humidity is read from the psychro-
these temperatures are denoted by Ta and Twb, the rate metric chart in the following way. The point on the
of heat transfer, Q, is given by Eq. (14), saturation curve corresponding to the wet-bulb tem-
perature is found first. An adiabatic cooling line is then
Q ¼ hAðTa
Twb Þ ð14Þ interpolated and followed until the coordinate corre-
sponding to the dry-bulb temperature is reached. The
Q ¼ rkg AðHi
Ha Þ ð15Þ humidity is read from the other axis.
Unit Processes in Pharmacy: Operations 3885

Humidification and Dehumidification for a certain time at the given temperature and then
quickly raising the temperature to the metastable
Most commonly, air is humidified by passage through region where further nucleation is negligible but the
a spray of water. Small quantities of air are easily already formed nuclei can grow.
dehumidified by adsorbing the water vapor with alu- Spontaneous nucleation occurs when sufficient
mina or silica gel arranged in columns. These are molecules of low kinetic energy come together in such
mounted in pairs so that one can be regenerated while a way that the attraction between them is sufficient to
the other is in use. Alternatively, the air can be cooled overcome their momentum.
below the dew point. Excess water vapor condenses
and the cold saturated air is reheated.
Crystal Growth

CRYSTALLIZATION If nucleation and crystal growth are independent, the

latter can be studied by seeding a melt with small crys-
The term crystallization describes the production of a tals under conditions of little or no natural nucleation.
solid, single-component, crystalline phase from a mul- The rate of growth can then be measured. The form of
ticomponent fluid phase. The importance of crystalli- the crystal growth curve is again explained by the
zation lies primarily in the purification achieved kinetics of the molecules. At temperatures just below
during the process and in the physical properties of the melting point, molecules have too much energy to
the product. A crystalline powder is easily handled, remain in the crystal lattice. As the temperature falls,
stable, and often possesses good flow properties and more molecules are retained and the growth rate
an attractive appearance. increases. Ultimately, however, diffusion to and orien-
Crystallization from a vapor, which occurs nat- tation at the crystal surface is depressed.
urally, e.g., in the formation of hoar frost, is employed For crystal growth in a single component melt, the
in sublimation processes and for the condensation of molecules at the crystal surface must reach the correct
water vapor during freeze-drying. position at the lattice and become suitably orientated,
losing kinetic energy. These energy changes appear as
heat of crystallization, which must be transferred from
Crystallization in Melts
the surface to the bulk of the melt. The rate of crystal
growth is influenced by the rate of heat transfer and the
A melt may be defined as the liquid form of a single
changes taking place at the surface. Agitation of the
material or the homogeneous liquid form of two or
system increases heat transfer by reducing the thermal
more materials which solidify on cooling. Crystalliza-
resistance of the liquid layers adjacent to the crystal
tion in such a system passes through the following
until the changes at the crystal face become the
stages: supercooling, nuclei formation, and crystal

Unit–Validation
controlling effect.
growth.
In multicomponent melts and solutions, deposition
If a single-component liquid is cooled, some degree,
of material at the crystal face depletes the adjacent
often high, of supercooling must be established before
liquid layers and a concentration gradient is set up
crystal nuclei form and growth begins. A metastable
with saturation at the face and supersaturation in the
liquid region exists below the melting point which only
liquid. Diffusion of molecules to the crystal face is dis-
can be entered by cooling. In this metastable, super-
cussed in the following section.
cooled region, the absence of nucleation precludes
the formation and growth of crystals. If, however, a
crystal seed is added, growth occurs. The deliberate
Crystallization from Solutions
seeding of a metastable system is commonly employed
in industrial crystallization. With further cooling,
During crystal growth, a high degree of supersatura-
spontaneous nucleation usually takes place and the
tion promotes a high growth rate. A reaction at the
released heat of crystallization raises the temperature
surface, in which, solute molecules become correctly
of the melt to its true melting point.
orientated in the crystal lattice, provides a second
resistance to the growth of the crystal. Simultaneously,
Nucleation the heat of crystallization must be conducted away.
For given conditions of temperature and saturation,
In certain single-component systems, such as piperine, agitation modifies the rate of crystal growth. Initially,
nucleation and crystal growth are independent and can agitation quickly increases the growth rate by decreas-
be separately studied. The rate of nucleation as a func- ing the thickness of the boundary layer and the diffu-
tion of supercooling is studied by maintaining the melt sional resistance. However, as agitation is intensified,
 3886 Unit Processes in Pharmacy: Operations

a limiting value is reached which is determined by the Once a particle is trapped at the entrance to the pore,
kinetics of the surface reaction. the contribution of the latter to the overall flow of
As with melts, soluble impurities may increase or liquid is very much reduced. Coarse straining with a
reduce nucleation rate. Insoluble materials may act as wire mesh and membrane filter employ this mech-
nuclei and promote crystallization. Impurities may also anism. Sterilization of liquids by filtration could be
affect crystal form and, in some cases, are deliberately regarded as an extreme application of clarification in
added to secure a product with good appearance, which the complete removal of particles as small as
absence of caking, or suitable flow properties. 0.3 mm must be ensured.

Crystallizers Cake filtration

Although other methods may be adopted, crystallizers The most common industrial application is the fil-
can be classified conveniently in the same way, a tration of slurries containing a relatively large amount
solution is supersaturated. This leads to the self- of suspended solids, usually 3 to 20%. The septum acts
explanatory terms, cooling crystallizer and evaporate only as a support in this operation, the actual filtration
crystallizer. In vacuum crystallizers, evaporation and being carried out by the solids deposited as a cake. In
cooling both take place. such cases, solids may completely penetrate the septum
until the deposition of an effective cake occurs. Until
this time, cloudy filtrate may be recycled. The physical
FILTRATION properties of the cake largely determine the method
employed. Washing and partial drying or dewatering
Filtration may be defined as the removal of solids sus- are often integral parts of the process. Effective dis-
pended in a liquid or gas by passage through a pervi- charge of the cake completes the process. The solids,
ous medium on which the solids are retained. The the filtrate, or both may be wanted.
pervious medium or septum is normally supported
on a base and these, together with a suitable housing The Theories of Filtration
providing free access of fluid to and from the septum,
comprise the filter. Filtration theory has two important aspects. The first
describes the flow of fluids through porous media
Methods and is applicable to both clarification and cake fil-
tration. The second, which is of primary importance
Clarification only in clarification, is the retention of particles on a
depth filter.
Unit–Validation

Clarification of parenteral solutions eliminates

unwanted solids normally present in very small con- Flow of fluids through porous media
centrations. This may be carried out with the help of
thick media, which allow the penetration and arrest The concept of a channel with a hydraulic diameter
of particles by entrapment, impingement, and electro- equivalent to the complex interstitial network which
static effects. This procedure leads to the concept of exists in a powder bed leads to Eq. (18),
depth filtration in which particles, perhaps a hundred
times smaller than the dimensions of the passages KA DP
Q ¼ ð18Þ
through the medium, are removed. Such filters are ZL
not absolute and must be designed with sufficient
depth so that the probability of the smallest particle where Q is the volumetric flow rate, A is the area of the
under consideration passing right through the filter is bed and L its thickness, DP is the pressure difference,
extremely small. and Z is the viscosity of the fluid. The permeability
Depth filtration fundamentally differs from the use coefficient, K, is given by
of media in which pore size determines the size of par-
ticle retained. Such filters may be said to be ‘‘absolute’’ e3
at a particle diameter closely related to the size of the 5ð1
eÞ2 S02
pore, so that there is a relatively sharp division between
particles which pass the filter and those that are where e is the porosity of the bed and S0 its specific
retained. An analogy with sieving may be drawn for surface area (cm2/cm3).
this mechanism. The life of such filters depends on In clarification, high permeability and filtration rate
the number of pores available for the passage of fluid. oppose good particle retention. In the formation of
Unit Processes in Pharmacy: Operations 3887

clarifying media from sintered or loose articles, accurate by particle size, size distribution, and shape in a man-
control of particle size, specific surface and porosity is ner described earlier. These factors may be varied for
possible, and, a medium can be designed which offers different filtering requirements.
the best compromise between permeability and particle Fixed media vary from perforated metals used for
retention. The analysis of permeability given above can coarse straining for the removal of very fine particles
be accurately applied to these systems. Because of with a sintered aggregate of metal, ceramic, plastic,
extremes of shape, this is not so with the fibrous media or glass powder. The size, size distribution, and shape
used for clarification. Here it is possible to develop a of the powder particles together with the sintering con-
material of high permeability and high retentive ditions control the size and distribution of the pores in
capacity. Such a material is, however, intrinsically weak the final product. The permeability may be expressed
and must be adequately supported. in terms of the coefficient given in Eq. (18). Alterna-
A mathematical account of the theories of clarifi- tively, the medium may be characterized by air
cation with depth filters is found in the work of Ives[4,5] permeability. The maximum pore size, which is impor-
and Maroudas and Eisenklam.[6] tant in the selection of filters for sterilization, may be
determined by measuring the pressure difference
required to blow a bubble of air through the medium
Filters while it supports a column of liquid with a known
surface tension.
The method by which the filtrate is driven through the
filter medium and cake, if present may be used to clas- Flexible media
sify filters into:
Flexible media may be woven or unwoven. Filter
 Gravity filters media, woven from cotton, wool, synthetic and regen-
 Vacuum filters erated fibers, and glass and metal fibers, are used as
 Pressure filters septa in cake filtration. Cotton is the most widely used
natural fiber, nylon is predominant among synthetic
Each group may be further subdivided into filters fibers. Terylene is a useful medium for acid filtration.
employed in continuous or batch processes although, Penetration and cake discharge are influenced by twist-
due to technical difficulties, continuous pressure filters ing and plying of fibers and by the adoption of various
are uncommon and expensive. Centrifugation is weaves such as duck and twill. The choice of a parti-
another means of removing filtrate. Extensive surveys cular cloth often depends on the chemical nature of
can be found in the literature.[7,8] the slurry.
Many small-scale filters simply consist of a fixed, Non-woven media in the form of felts and com-
rigid medium, robust enough to withstand limited pressed cellulose pulps, are used for clarification by
pressures, mounted in a suitable housing. These filters,

Unit–Validation
depth filtration. Unless carefully prepared, they have
which are also vacuum operated, are used to clarify by the disadvantage of losing fibrous material from the
depth filtration. Media are composed of sintered downstream side of the filter. The application of sheet
metals, ceramics, plastics, or glass. Filters prepared media has been discussed earlier. High wet strength
from closely graded and sintered chemical powders is conferred on paper sheets by resin impregnation.
are suitable for the sterilization of solutions by An alternative technique employs asbestos fibers
filtration on a manufacturing scale. supported in a cellulose framework.

Mechanism of air filtration

Filter Media
A theoretical foundation for the filtration of air by
In cake filtration, the medium must oppose excessive passage through fibrous media was laid in the early
penetration and promote the formation of a junction 1930s by studies of the flow of suspended particles
with the cake, to high permeability. The medium around various obstacles. In studies of the filtration
should also give free discharge of cake after washing of smokes[9,10] it has been shown that the following fac-
and dewatering. tors operate simultaneously in the arrest of a particle
during its passage through a filter, although their rela-
Rigid media tive importance varies with the type of filter and the
conditions under which it is operated.
Rigid media may be loose or fixed. The former is exem-
plified by the deposition of a filter aid on a suitable  Diffusion effects due to Brownian movement.
support. Filtration characteristics are governed mainly  Electrostatic attraction between particles and fibers.
 3888 Unit Processes in Pharmacy: Operations

 Direct interception of a particle by a fiber. filter thickness is capable of retaining 90% of the enter-
 Interception as a result of inertial effects acting on a ing particles and 106 particles enter, 105 penetrate. If
particle and causing it to collide with a fiber. six thicknesses are used, Eq. (21) predicts that only
 Settling and gravitational effects. one particle penetrates. The log-penetration effect has
been confirmed for fibrous filters[13] and granular
Air filters operate under conditions of streamline beds.[18]
flow as indicated by the streamlines drawn around a
cylindrical fiber. It was assumed that capture of a par-
ticle takes place if any contact is made during its move- Centrifugal Operations
ment around the fiber. Once captured, the particle is
not re-entrained in the air stream and deposited deeper An object moving in a circular path is subjected to an
in the bed. Support for this assumption has been found outward centrifugal force which balances the centrip-
by using an atomized suspension of Staphylococcus etal force moving the object toward the center of
albus and spores of Bacillus subtilis.[11] Nevertheless, rotation. This principle is used in the mechanical
some fiber filters are treated with viscous oils, presum- separations called centrifugal filtration and centrifugal
ably to make capture more positive and to reduce sedimentation. In the former, a material is placed in a
re-entrainment. rotating perforated basket which is lined by a filter
Deviation of particles from streamlines can occur cloth used to separate a solid, which is retained at
ina number of ways.[10,12] The chance of capture the cloth, from a liquid. It is essentially a filtration pro-
increases if Brownian movement causes appreciable cess in which the driving force is of centrifugal origin.
migration across streamlines, an effect only important This does not depend upon a difference in the density
for small particles (less than 0.5 mm) and low airspeeds, of the two phases.
when the time span spent in the vicinity of a fiber is In centrifugal sedimentation, the separation is due
relatively long. These conditions also apply to capture to the difference in the density of two or more phases.
which is the result of electrostatic attraction. This is the more important process, where both solid–
Sampling efficiency has been demonstrated for bac- liquid mixtures and liquid–liquid mixtures can be com-
terial aerosols[13] in a study of the efficiency with which pletely: separated. If, however, the separation is incom-
a glass fiber mat collected B. subtilis spores atomized plete, there is a gradient in the size of the dispersed
as particles just over a micrometer in radius. A theo- phase within the centrifuge due to the faster radial
retical approach to the removal of industrial dusts has velocity of the larger particles. Operated in this way,
been developed.[14–16] the centrifuge becomes a classifier.

Design, operation, and testing of air filters Centrifugal sedimentation

Unit–Validation

Granular beds, fibrous media, and ‘‘absolute filters’’ The motion of a particle in a liquid is described by
prepared from cellulose and asbestos are used for Stokes’ equation. If its diameter is d, the rate u at
high-efficiency air filtration. With fibrous and granular which it settles by gravity in a liquid of viscosity Z
filters, the fractional reduction in particle content is and density r is given by Eq. (21)
assumed to be the same through successive incremental
1 2 rs
r
thicknesses of the filter, expressed by Eq. (19), u d g ð21Þ
18 Z
dC
¼
kC ð19Þ where g is the acceleration due to gravity, and rs is the
dx
density of the particle. In the centrifuge, the gravi-
where C represents the number of particles entering a tational force causing separation is replaced by a cen-
section of thickness dx. The constant, k, is a measure trifugal force. If the particle has a mass m and moves
of the filter’s ability to retain a particle. It is a complex at an angular velocity o in a circle of radius r, the cen-
function of fiber diameter, interfiber distance, and the trifugal force is o2r  (m
m1), where m1, is the mass
operational air velocity. Integration between inlet and of the displaced ligand. The expression
outlet conditions gives Eq. (20). o2 r
Cout g
log ¼
kC ð20Þ
Cin
is, therefore, the ratio of the centrifugal and gravi-
The use of this log penetration effect in filter tational forces in the example described previously.
design has been described elsewhere.[17] If a certain Its value can exceed 10,000. The separation is quicker,
Unit Processes in Pharmacy: Operations 3889

more complete, and effective in systems containing humidity, wet-bulb temperature, and adiabatic cooling
very fine particles which do not settle by gravity line. Other terms may be defined as:
because of Brownian movement.
Expressing the mass of the particle in terms of its  Moisture Content. It is usually expressed as weight
volume and effective density, the centrifugal force per unit weight of dry solids.
can be written as in Eq. (22).  Equilibrium Moisture Content. If a material is
exposed to air at a given temperature and humidity,
p 2 it will gain or lose moisture until equilibrium is
d ðrs
rÞo2 r ð22Þ
6 reached. The moisture present at this point is
defined as the equilibrium moisture content for
In streamline conditions, the opposing viscous force
the given exposure conditions. At a given tempera-
is 3p dZu, where u is the terminal velocity of the par-
ture, it will vary with the partial pressure of the
ticle. Equating these expressions gives Eq. (23).
water vapor in the surrounding atmosphere.
1 2 ðrs
rÞ 2
u ¼ d o r ð23Þ Equilibrium moisture content curves vary greatly
18 Z
with the type of material examined. Insoluble, non-
The rate of sedimentation is proportional to the porous materials, such as talc or zinc oxide, have equi-
radius of the basket and the square of the speed librium moisture contents of almost zero over a wide
at which it rotates. Centrifugal sedimentors can be humidity range. A moisture content between 10 and
divided into a number of types. For operations at very 15% may be expected for cotton fabrics under; normal
high speeds, the centrifuge bowl is tubular with a atmospheric conditions. Drying below the equilibrium
length/diameter ratio from 4 to 8. The solids are moisture content for room conditions may be deliber-
periodically discharged by scraping the walls of the ately undertaken, particularly if the material is
centrifuge tube. Uses include the cleaning of fats and unstable in the presence of moisture; subsequent sto-
waxes, the fractionation of blood, and the recovery rage becomes important.
of viruses. The effects of storage after drying also may be
assessed from the equilibrium moisture content curves.
Storage conditions are not critical for the lactose
DRYING granulation.[19,20] If the antacid formulation is stored
at a relative humidity of only 65% it would, given
Drying may be defined as the vaporization and sufficient time, absorb moisture until the content was
removal of water or other liquid from a solution, sus- 91%. This could be associated with poor flow character-
pension, or other solid–liquid mixture to form a dry istics and its attendant difficulties during compression.
solid. The change of phase from liquid to vapor distin-

Unit–Validation
guishes drying from the mechanical methods of sepa-
rating solids from liquids such as filtration. The latter Evaporation of Water into an Air Stream
often precede drying because they offer a cheaper
method for removing a large part of the liquid, where The evaporation of moisture into a warm air stream,
applicable. with the latter providing the latent heat of evaporation,
Adjustment and control of moisture levels by dry- is a common drying mechanism although it is not
ing, is important in the manufacture and development easily adapted to the recovery of the liquid. In the
of pharmaceutical products. Apart from the obvious evaporation from a liquid surface which, with the
requirement of dry solids for many operations, drying passage of air, falls to the wet bulb temperature corre-
may be carried out in order to: sponding to the temperature and humidity of the air,
the rate at which water vapor is transferred from the
 Improve handling characteristics, as in bulk powder saturated layer at the surface to the drying stream is
filling and other operations involving powder flow, described by Eq. (15),
and
 Stabilize moisture-sensitive materials, such as kg
N ¼ ðPwi
Pwa Þ
aspirin and ascorbic acid. RT

where Pwi is the partial pressure of the water vapor at

Theory the surface, Pwa is the partial pressure of water vapor in
the air, kg is a mass transfer coefficient, and N the num-
The following terms are employed in discussing drying: ber of moles vapor transferred from unit area in unit
humidity and humidity of saturated air, relative time. Rewriting this in terms of the total mass, W,
 3890 Unit Processes in Pharmacy: Operations

transferred in unit time from the entire drying surface A B

A gives Eq. (6), A B A

Moisture content

Rate of drying
Mw A
W ¼ kg ðPwi
Pwa Þ
RT C
B
C D
where Mw is the molecular weight of water vapor, R is D
the gas constant, and T the absolute temperature. The
mass transfer coefficient, kg, is a function of the tem- Time Moisture content
perature, the air velocity, and the angle of air inci- Fig. 5 (A) Relation of moisture content and time of drying;
dence. A high velocity or angle of incidence (B) rate of drying and moisture content.
diminishes the thickness of the stationary air layer in
contact with the liquid surface and therefore lowers
the diffusional resistance. The initial heating period during which equilibrium is
The rate of evaporation may also be expressed in established is short and has been omitted from both
terms of the heat transferred across the laminar film figures. Assuming that sufficient moisture is initially
from the drying gases to the surface, as shown in present, the drying-rate curve exhibits three sections
Eq. (9), limited by the points A, B, C, and D. In section A-B,
called the constant-rate period, moisture is evaporating
Q ¼ hAðTa
Ts Þ from a saturated surface at a rate governed by the
stationary air film in contact with it. An analogy with
where Q is the rate of heat transfer, A is the area of the evaporation from a plain water surface can therefore
surface, Ta and Ts, are the temperatures of the drying be drawn and Eqs. (11) and (13) apply. The rate of dry-
air and the surface, respectively, and h is the heat trans- ing during this period depends upon the air tempera-
fer coefficient. The last is also a function of air velocity ture, humidity, and speed, which in turn determine
and the angle of impingement. If the latent heat of the temperature of the saturated surface. Assuming
evaporation is l, this affords a mass transfer rate, W, that these are constant, all variables in the drying equa-
which is given by Eq. (24). tions given earlier are fixed, and a constant rate of dry-
ing is established which is largely independent of the
hA material being dried. The drying rate is somewhat
W ¼ ðTa
Ts Þ ð24Þ
l lower than for a free-water surface and to some extent
depends on the particle size of the solids. During the
When these conditions pertain to drying, the surface
constant-rate period, liquid must be transported to
temperature, Ts, which is the wet bulb temperature, is
the surface at a rate sufficiently high to maintain satu-
normally much lower than the temperature of the dry-
Unit–Validation

ration. The mechanism of transport is discussed later.

ing gases. This is of great importance in the drying of
At the end of the constant-rate period B, a break in
thermolabile materials. If solids are present in the sur-
the drying curve occurs. This point is called the critical
face, the rate of evaporation is modified, the overall
moisture content, and a linear fall in the drying rate
effect depending on the structure of the solids and
occurs with further drying. Because section, B-C, is
the moisture content.
called the first falling-rate period. At and below the
critical moisture content, the movement of moisture
Static Beds of Non-porous Solids from the interior is no longer sufficient to saturate
the surface. As drying proceeds, moisture reaches the
The drying of wet granular beds containing non- surface at a decreasing rate and the mechanism which
porous particles, which are insoluble in the wetting controls its transfer influences the rate of drying.
liquid, has been extensively studied. The operation is Because the surface is no longer saturated, it tends to
presented as the relation of moisture content and time rise above the wet bulb temperature.
of drying in Fig. 5A. It should be noted that the For any material, the critical moisture content
equilibrium moisture content is approached slowly. A decreases as the particle size decreases. Eventually,
protracted period may be required for the removal of moisture ceases to reach the surface which becomes
water just above the equilibrium value. This is not dry. The plane of evaporation recedes into the solid,
justified if a small amount of water can be tolerated the vapor reaching the surface by diffusion through
in further procession establishing realistic drying the pores of the bed. This section is called the second
requirements. falling-rate period and is controlled by vapor diffusion,
The data have been converted to a curve relating a factor which is largely independent of the conditions
the rate of drying to moisture content in Fig. 5B. outside the bed but markedly affected by the particle
Unit Processes in Pharmacy: Operations 3891

size due to its influence on the dimensions of pores and Through Air-Circulation Drying
channels. During this period, the surface temperature
approaches the temperature of the drying air. If the particles are in a suitable granular form, it is
Considerable migration of liquid occurs during the often possible to pass the air stream downward
constant-rate and first falling-rate periods. Associated through the bed of solids. Drying follows the pattern
with the liquid is any soluble constituent which forms described previously, except that each particle or
a concentrating solution in the surface layers as agglomerate behaves as a drying bed. The surface area
drying proceeds. Deposition of these materials takes exposed to the drying gases is greatly increased, and
place when the surface dries. Considerable segregation the drying rates are 10 to 20 times higher than those
of soluble elements in the cake can occur, therefore, encountered when air is passed over a free surface.
during drying. These effects have been fully investi-
gated.[21]
Methods Involving Movement of the Solid

As an extension of drying by passing the air stream

The Internal Mechanism of Drying through a static bed of solids, it is possible to project
air upward through the bed at a velocity high enough
Capillary forces offer a coherent explanation for the to fluidize the particles. Alternatively, the material may
drying periods of many materials. If a tapered capillary be mechanically subdivided and introduced into the
is filled with water and exposed to a current of air, the drying stream. Both methods give high drying rates
meniscus at the smaller end remains stationary while due to high interfacial contact between the drying sur-
the tube empties from the wider end. A similar situa- faces and the air stream. Fluidized bed driers and spray
tion exists in a wet particulate bed and the phenom- driers, respectively, are based on these principles.
enon is explained by the concept of suction potential.
A negative pressure exists below the meniscus of a
curved liquid surface which is proportional to the sur- Solids Moving Over a Hot Surface
face tension, l, and inversely proportional to the radius
of curvature, r. (The meniscus is assumed to be a part Conditions in which the solids move over a heated
of a hemisphere.) This negative pressure or suction surface are employed in tumbling and agitated driers.
potential may be expressed as the height of liquid, Drying rates are higher than those obtained in static
expressed by Eq. (25), beds because fresh solids are continually exposed to
the hot surface. The heat treatment received by the
solid is more uniform.
2l
h ¼ ð25Þ
rgr

Unit–Validation
Batch Driers

where r is the density of the liquid. Hot air ovens

The suction potential, hx, acting at a depth x below
the meniscus is given by Eq. (26). Ovens operating by passing hot air over the surface of
a wet solid which is spread over trays arranged in
racks, provide the simplest and cheapest drier. In small
hx ¼ h
x ð26Þ
installations, the air is passed over electrically heated
elements and once through the oven. Larger units
The particles of the bed enclose spaces called pores may employ steam-heated, finned tubes, and thermal
connected by passages, the narrowest part of which is efficiency is improved by recirculating the air. This is
called the waist. The dimensions of the latter are controlled by manually set dampers, and a common
determined by the size of the surrounding particles operating position gives 90% recirculation and 10%
and the manner in which they are packed. In a ran- bleed-off. The heater blank is placed in such a position
domly packed bed, pores and waists of varying sizes that the solids do not receive radiant heat and
are found. Thus, the radius of a capillary running incoming air may be filtered.
through the bed varies continuously. The depletion The chief advantage of the hot air oven, apart from
of water in this network is controlled by the waists its low initial cost, is its versatility. With the exception
because the radii of curvature are smaller and the suc- of dusty solids, materials of almost any physical form
tion potentials are greater than for the pores. The may be dried. Thermostatically controlled air tempera-
application of this mechanism has been described fully tures between 40 and 120 C permit heat-sensitive
elsewhere.[22,23] materials to be dried. For small batches this may be
 3892 Unit Processes in Pharmacy: Operations

the equipment of choice. However, the following char- possibility of solvent recovery, and increased drying
acteristics have led to development of other small rates. Heat is supplied to the tumbling charge by con-
driers: tact with the heated shell and heat transfer through the
vapor.
 A large floor space is required for the oven and tray A normal charge would be about 60% of the total
loading facilities. volume and, for driers 2–7 ft (0.6–2 m) in diameter,
 Labor costs for loading and unloading the oven are drying times of 2–12 h may be expected. In studying
high. the application of tumbler driers to drying tablet gran-
 Long drying times, usually of the order of 24 h, are ules, periods of 2–3.5 h were sufficient instead of 18 h
necessary. required by hot air ovens.[25] The mixing and granulat-
 Solvents can be recovered from the air only with ing capacity of the tumbling action has suggested that
difficulty. these operations could precede drying in the same
 Unless carefully designed, non-uniform distribution apparatus.
of air over the trays results in variations in tempera-
ture and drying times within the oven. Variations of Fluid-bed driers
7 C in temperature have been found from
location to location during the drying of tablet The term ‘‘fluidization’’ is applied to processes in
granules.[19] Poor air circulation may permit local which a loose, porous bed of solids is converted to a
saturation and the cessation of drying. fluid system, having the properties of surface leveling,
flow, and pressure-depth relationships, by passing the
An extensive analysis of tray drying and the effect of fluid up through the bed.
operational variables has been given by Shepherd, Fluidized-bed techniques, employing air as the flui-
Hadlock, and Brewer.[24] dizing medium, have been successfully applied to the
If the material is of suitable granular form, drying drying of solids of the suitable physical form. The high
times may be reduced to 1 h or less by passing the interfacial contact between drying air and solids gives
air downward through the material laid on mesh drying rates 10 to 20 times higher than those obtained
trays. The oven in this form is called a batch through- during tray drying.
circulation drier. Fluidized-bed driers are particularly suitable for
granulated materials and are being increasingly used
Vacuum tray driers for tablet granulations, providing that product change-
over is not too frequent. Machines vary in size, handl-
Vacuum tray driers offer an alternative method for ing up to 250 kg. Drying times, maximum, minimum,
drying small quantities of material. When scaled up, and optimum air velocities, air temperature, and the
construction becomes massive to withstand the applied tendency to cake and channel are established exper-
Unit–Validation

vacuum and cost is further increased by the associated imentally as those cannot be predicted accurately at
vacuum equipment. Vacuum tray driers are, therefore, present.
only used when a definite advantage over the hot air
oven is secured, such as low temperature drying of
thermolabile materials or the recovery of solvents from Agitated batch driers
the bed. The exclusion of oxygen may also be advan-
tageous or necessary in some operations. Agitated batch driers consist of a jacketed cylindrical
Heat is usually supplied by passing steam or hot vessel with agitator blades designed to scrape the bot-
water through hollow shelves. Radiation from the shelf tom and walls. They may operate at atmospheric pres-
above may cause a significant increase in temperature sure or under vacuum. Pasty materials that could not
at the surface of the material if high drying tempera- be handled in tumbling or fluidized-bed driers, may be
tures are used. Drying times are long, usually of the successfully dried at rates higher than can be achieved
order of 12 to 48 h. in an oven.

Tumbling driers Freeze drying

The limitations of ovens, particularly with respect to Freeze drying is an extreme form of vacuum drying in
the long drying times, has, where possible, promoted which the solid is frozen and drying takes place by sub-
the design and application of other batch driers. The liming the solid phase[26–30] at low temperatures and
simplest of these is the tumbler drier. Its most common pressures. Establishing and maintaining these con-
shape is the double cone.[19] Operating under vacuum, ditions, together with the low drying rates obtained,
it provides controlled low temperature drying, the constitutes the most expensive method of drying which
Unit Processes in Pharmacy: Operations 3893

is only used on a large scale when other methods are the drying gas is cooler and the material is almost
inadequate. dry, a condition in which many materials are thermally
Freeze drying is extensively used when rapid less sensitive.
decomposition occurs during normal drying. Another Drying is considered to take place by simple evapor-
application concerns substances that can be dried at ation rather than by boiling and it has been observed
high temperatures but are thereby changed in some that a droplet reaches a terminal velocity within about
way. one foot of the atomizer. Beyond this, there is no rela-
Freeze drying is theoretically a simple technique. tive velocity between the droplet and the drying gas
Pure ice exhibits an equilibrium vapor pressure of unless the former is very large. The droplets may dry
4.6 mm Hg (611 Pa) at 0 C and 0.1 mm Hg (13.3 Pa) to form a solid, spherical particle. If, however, the
at
40 C. The vapor pressure of ice containing dis- emerging solids form a skin, internal pressure may
solved substances is, of course, lower. If, however, inflate the particle and the final dry form will consist
the pressure above the frozen solution is less than its of hollow spheres which may or may not have a blow
equilibrium vapor pressure, the ice sublimes, eventually hole. These xenospheres may also fragment, resulting
leaving the solute as a sponge-like residue equal in in a final product of agglomerates of finely divided
apparent volume to the original solid. solids.
The capital and running costs of spray driers are
high, but if the scale is sufficiently large, it may provide
Continuous Driers the cheapest method. When thermolabile materials are
dried on a small scale, costs will be 10 to 20 times
Although many types of continuous driers are avail- higher than for oven drying. Air used to dry fine che-
able, the scale of the operation for which they are micals or food products is heated indirectly, thus
designed is rarely appropriate to pharmaceutical reducing thermal efficiency and increasing costs. In
manufacture. As with most continuous operations, some other installations, hot gases from combustion
the cost is disproportionately high for small units. may be used directly.
Spray and drum driers provide an exception, because
residence times in the driers are short and thermal Drum driers
degradation is minimized. Under some conditions,
freeze drying may be the only practicable alternative. The drum drier consists of one or two slowly rotating,
steam-heated cylinders. These are coated with solution
Spray driers or slurry by means of a dip feed in which the lower por-
tion of the drum is immersed in an agitated trough of
The solution or suspension to be dried is sprayed into a feed material or, in the case of some double-drum
hot air stream and circulated through a chamber. The driers, by feeding the liquor into the gap between the

Unit–Validation
dried product may be carried out to cyclone or bag cylinders. Spray and splash feeds are also used. In
separators or may fall to the bottom of the drying dip feeding, the hot drum must not boil the liquid in
chamber and be expelled through a valve. The cham- the trough. Drying takes place by simple evaporation
bers are normally cylindrical with a conical bottom rather than by boiling. The dried material is scraped
although proportions vary widely. The process can from the drum by a knife at a suitable point.
be divided into four sections: atomization of the fluid, Drying capacity is influenced by the speed of the
mixing of the droplets, drying, and finally removal and drum and the temperature of the feed, which may be
collection of the dry particles. preheated. With the double-drum drier, the gap
In vertical spray driers, the flow of the drying gas between the cylinders determines the thickness of
may be concurrent or counter-current with respect of the film.
the movement of droplets. The movement of the Drum driers, like spray driers, are relatively
gas is, however, complex and highly turbulent. Good expensive in small sizes and their use in the pharmaceu-
mixing of droplets and gas occurs, and the heat and tical industry is largely confined to drying thermo-
mass transfer rates are high. In conjunction with the labile materials where the short contact time is
large interfacial area conferred by atomization, these advantageous. Drums are normally fabricated from
factors give very high evaporation rates. The residence stainless or chrome-plated steel to reduce contami-
time of a droplet in the drier is only a few seconds (5– nation. The heat treatment to which the solid is sub-
30 s). Since the material is at wet bulb temperature for jected is more intense than in spray drying and the
much of this time, high gas temperatures of 150–200 C physical form of the produce is often less attractive
may be used even with thermolabile materials. during drying, the liquid approaches its boiling point
Although the temperature of the material rises above and the dry solids attain the temperature of the drum
the wet-bulb temperature at the end of the process, surface.
 3894 Unit Processes in Pharmacy: Operations

SIZE REDUCTION AND CLASSIFICATION crushing, the application of stress continues on the
crushed bed of particles. Although further size reduction
The theoretical strength of crystalline materials can be occurs, the process is less efficient due to vitiation
calculated from interatomic attractive and repulsive of energy by the effects of interparticulate friction
forces. The strength of real materials, however, is and stress transmission via particles which do not
found to be many times smaller than the theoretical themselves fracture. This is easily demonstrated when a
value. The discrepancy is explained in terms of flaws crystalline material is ground in a pestle and mortar.
of various kinds, such as minute fissures or irregulari- The fine powder initially produced protects coarser
ties of lattice structure known as dislocations. These particles. If the material is sieved and oversize particles
have the capacity to concentrate the stress in the vicin- are returned, the operation may be completed with far
ity of the flaw. Failure may occur at a much lower less effort.
overall stress than is predicted from the theoretical Various hypotheses relate the net grinding energy
considerations. Failure occurs with the development applied to a process and the size reduction achieved.
of a crack tip which propagates rapidly through the The first, proposed by Karl von Rittinger in 1867,
material, penetrating other flaws which may, in turn, states in Eq. (27) that the energy necessary for size
produce secondary cracks. The strength of the material reduction is directly proportional to the increase in sur-
depends therefore on the random distribution of flaws face area,
and is a statistical quantity varying within fairly side
limits. This concept explains why a material becomes E ¼ kðSp
Sf Þ ð27Þ
progressively more difficult to grind. Since the prob-
ability of containing an effective flaw decreases as the
particle size decreases, the strength increases until, with where E is the energy consumed, and Sp and Sf are the
the achievement of faultless domains, the strength of surface area of the product and feed materials, respec-
the material equals the theoretical strength. This tively. The constant, k, depends on the grinding unit
position is not realized in practice due to complicating employed and represents the energy consumed in
factors such as aggregation. enlarging the surface area by one unit. The relation
The strength of most materials is greater in com- between surface area and particle size has already been
pression than in tension. It is therefore unfortunate derived, and Eq. (28) may therefore be written,
that technical difficulties prevent the direct application

1 1
of tensile stresses. The compressive stresses commonly E ¼ k
ð28Þ
used in comminution equipment do not cause failure dp df
directly but generate by distortion sufficient tensile or
shear stress to form a crack tip in a region away from where df and dp are the particle sizes of feed and pro-
the point of primary stress application. This is an inef- duct particles, respectively.
Unit–Validation

ficient but unavoidable mechanism. Impact and The hypothesis indicates that energy consumption
attrition are the other basic modes of stress appli- per unit area of new surface produced increases faster
cation. The distinction between impact and com- than the linear ratio of feed and product dimensions, a
pression is referred to later. Attrition, which is phenomenon already noted and explained. The pro-
commonly employed, is difficult to classify but is prob- portionality of net energy input and new surface pro-
ably primarily a shear mechanism. duced has been confirmed in some grinding operations.
The deformation and subsequent failure of a brittle Conversion of grinding energy to surface energy is
material is not only a function of stress but also of the neglected in Kick’s law, promulgated in 1885. It is
rate at which the stress is applied. Different results may based on the deformation and brittle failure of elastic
be obtained from slow compressive breaking and bodies and states that the energy required to produce
impact breaking at the same energy level. Particle analogous changes of configuration of geometrically
shape, size, and size distribution may be affected. In similar bodies is proportional to the weight or volume
impact breaking, the rate of stress application is so of those bodies. The energy requirements are inde-
high that the limiting strain energy may be exceeded pendent of the initial particle size and depend only
several times by the suddenness of the operation. The on the size reduction ratio. Kick’s law predicts lower
reason is that fracture is time dependent, a lag occur- energies than the relation proposed by Rittinger. The
ring between the application of maximum stress and theory, however, demands that the resistance to crush-
failure. ing does not change with particle size. The role of flaws
Stress application is further complicated by ‘‘free present in real materials is not considered, with the
crushing’’ and ‘‘packed crushing’’ mechanisms. In free result that the energy required for fine grinding, when
crushing, the stress is applied to an unconstrained the apparent strength may have greatly risen, is under-
particle and released when failure occurs. In packed estimated.
Unit Processes in Pharmacy: Operations 3895

A third theory of comminution gives results inter- which they were not designed. A limited size reduction
mediate between the predictions of the laws of Kick ratio is imposed upon a single operation, larger ratios
and Rittinger.[31] It rests upon three principles: the first being obtained by the adoption of several stages, each
states that any divided material must have a positive employing a suitable mill. The fluid energy mill,
energy register. This can only be zero when the particle which presents a size reduction ratio of up to 400, is
size becomes infinite. The input energy, E, for any size exceptional.
reduction process then equals the product energy A low retention time is inherent in free crushing
register minus the feed energy register. The energy machines. Little overgrinding takes place and the pro-
associated with a powder increases as the particle size duction of excessive undersize material or fines is
decreases, and it may be assumed that the energy regis- avoided. Protracted milling times are found with many
ter is inversely proportional to the particle size to an low-speed mills, with the result that considerable over-
exponent, n. Hence, Eq. (29) is valid. grinding takes place. Accumulation of product parti-
cles within the mill reduces the effectiveness of
K K breaking stresses and the efficiency of milling progress-
E ¼ Ep
Ef ¼
n ð29Þ ively decreases.
dnp df
Dry and wet grinding
The second principle of this theory[31] assigns to n a
value of i, stating that ‘‘the total work useful in break- Between the approximate limits of 5 and 50% moisture,
ing, which has been applied to a stated weight of an materials cake and do not flow. Both factors oppose
homogeneous material, is inversely proportional to effective grinding. Dry grinding is carried out at low
the square root of the diameter of the product parti- moisture contents, the upper limit depending on the
cles.’’ nature of the material. Although 5% or more moisture
The third principle states that breakage of the may be permissible for vegetable drugs, it would prove
material is determined by the flaw structure. This excessive during the milling of a coarse, impervious
aspect of size reduction has already been discussed. solid.
A modification to Kick’s law, sometimes known as Wet grinding is a common procedure when a fluid
the fourth law of comminution, has also been pro- suspension is required and drying, which would pro-
posed.[32] For its discussion, the reader is referred to vide a significant drawback, is unnecessary. An excel-
the original paper. lent dispersion can be produced simultaneously,
An empirical, but realistic approach to mill which in some operations provides the primary objec-
efficiency is gained through experiments in which the tive, size reduction being of secondary importance.
energy consumed and size reduction achieved are com- Wet grinding also may be adopted when the size
pared with values obtained in a laboratory test operat- reduction achieved during dry grinding is prematurely
ing under free crushing conditions. All energy supplied

Unit–Validation
linked by aggregation.
in the latter is available for crushing and the test is Certain general advantages are secured during wet
assumed to be 100% efficient. Both slow crushing grinding, including increased mill capacity, a lower
and impact tests are used. A large number of single energy consumption, the elimination of hazards from
particles may be simultaneously crushed and the work dust, and easier handling of materials. The principal
done is measured.[33] The latter is related to the size disadvantage, apart from the possible inclusion of a
reduction. Similar measurements can be made during drying stage, is the increased wear of the grinding
practical milling, expressing the efficiency of the pro- medium.
cess as a percentage of the free crushing value. On this
basis, the approximate efficiency of the roll crusher is Temperature sensitivity
80%, of the swing hammer mill 40%, of the ball mill
10%, and of the fluid energy mill only 1%. Care must be exercised during the milling of tempera-
ture-sensitive materials, especially for a very fine pro-
duct; caking results if the softening point is exceeded.
The Operation of Mills Materials may be chilled before grinding or facilities
provided for cooling the mill during grinding. Waxy
Heywood[34] has stated that any type of crushing or solids can be successfully ground with dry ice, the
grinding machine exhibits optimal comminution con- low temperatures conferring brittle characteristics on
ditions for which the ratio of the energy to new surface the material. Chemical degradation may occur at high
is minimal. If finer grinding is attempted in such a grinding temperatures. Oxidative changes can be pre-
machine, the; ratio is increased. Mills may thus become vented by grinding in an inert atmosphere such as
grossly inefficient if called upon to grind at a size for nitrogen.
 3896 Unit Processes in Pharmacy: Operations

Structural changes usually short and many undersize particles traverse it

without falling through. With an increase of sieving
Several examples of change of physical structure dur- area, the meshes become more fragile and the finest
ing very fine grinding have been reported, for example, meshes must be supported with a coarser wire. An
changes in the crystal form of calcium carbonate after example of a large-scale separator utilizes a circular
ball milling,[35] distortion of the kaolinite lattice,[36] screen, up to 5 ft (1.5 m) in diameter, vibrated in a hori-
and formation of various barbiturate polymorphs.[37] zontal plane, the gyratory movement being imparted
Changes such as these could affect solubility and other by an out-of-balance fly wheel connected to the
physical characteristics which, in turn, might influence assembly. In other machines, the mesh is rectangular
formulation and therapeutic value. and inclined at a shallow angle (5–30 ). A gyratory
movement is developed and the material to be classi-
Dust hazards fied is fed to the top. These machines may bear more
than one deck, thus allowing the separation of the
Hazards from dust may become acute during dry powder into several fractions at one time.
grinding. Extremely potent materials require dust
proofing of machines and dust-proof clothing and Elutriation and sedimentation
masks for operators. Danger may also arise from the
explosive nature of many dusts. The simplest classifier is a rising current of fluid in
which the particles are suspended. In this case, the
Grinding Equipment force opposing the upward drag is gravitational. If
the opposition develops a terminal velocity higher than
The following equipment are in regular use for dry- the current speed, the particle falls. This is the principle
grinding pharmaceutical materials: edge- and end-runner of elutriation; the particle size d at which the sepa-
mills, hammer mills, pin mills, ball mills, vibratory mills, ration is made follows from a rearrangement of
fluid energy mills, colloid mills, and roller mills. Eq. (29) for conditions in which Stokes’ law is valid;
it is given by Eq. (30),
sffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
Classification or Size Separation 18Zm
d ¼ ð30Þ
ðrs
rÞg
Although a number of particle properties can be used
to classify a powder, only two are important. The first
is based on the ability of a particle to pass through an where rs
r is the density difference between solid
aperture. This is sieving or screening. The second and fluid, Z is the viscosity of the fluid, and m is the
employs the drag forces on a particle moving through speed of the upward current.
In practice, fluctuations in flow conditions due to
Unit–Validation

a fluid. The term ‘‘classification’’ is sometimes restricted

to this method of separation but here the terms ‘‘elutria- natural convection and a violation of the conditions for
tion’’ and ‘‘sedimentation’’ are used. In general, screen- which Stokes’ law is valid, blur the point of separation.
ing is applied to the separation of coarse particles and The centrifuge is normally operated to completely
sedimentation to the separation of fine particles. separate two phases. If, however, the rate at which
the feed passes through does not allow all particles to
Sieving and screening settle, the action of a classifier is developed. This is
illustrated by a solid-bowl centrifuge which consists
Sieves and screens are widely used for the classification of a steel shell in the form of a frustum mounted
of relatively coarse materials. For very large particles horizontally. It contains a conveying screw at the wall
(>0.5 in.) a robust plate perforated with holes is used. which rotates at a slightly higher speed than the shell.
However, the pharmaceutical applications of screening Particles that settle at the wall are conveyed to the
are for much smaller particles and screens are in the narrow end of the shell and discharged. Fine particles
form of woven meshes. Unless special methods are are entrained with the overflow to the other end.
used to prevent clogging and powder aggregation, the Further details of this and other centrifugal classifiers
lower useful limit is in a cloth woven with 200 mesh/in. have been given by Treasure.[38]
(70–80 mm). Fine screens of this type are extremely fragile
and must be used with great care.
As the scale of the operation increases it becomes, in MIXING
general, less precise. For continuous screening, the feed
material is made to move across the screen to a point Mixing has been defined[39] as an operation in which
of discharge. The residence time on the screen is two or more ingredients in separate or roughly mixed
Unit Processes in Pharmacy: Operations 3897

condition are treated so that each particle of any one number of particles in a sample. If the particle size is
ingredient is adjacent to a particle of each of the other reduced to the extent that the same weight of sample
ingredients, as nearly as possible. The term ‘‘blending’’ contains four times as many particles, the standard
is synonymous and ‘‘segregation’’ or ‘‘demixing’’ is the deviation is halved.
opposite. In a critical examination of pharmaceutical mixing,
Mixing has been classified[40] as follows: Train[42] showed that samples of a random mixture of
equal parts A and B must contain at least 800 particles
 Positive mixing which applies to systems that, given if 997 out of every 1000 samples were to lie between
time, would spontaneously and completely mix. 10% of the stated composition, that is, the pro-
Examples are provided by two gases or two miscible portion, p, of A ¼ 0.5  0.05, where s ¼ 0.05/3. If
liquids; mixing apparatus is used on such systems to limits of 1% were substituted, 90,000 particles must
accelerate mixing. be present in each sample. The true standard deviation
 Negative mixing is demonstrated by suspensions of is given by s. The standard deviation estimated by the
solids in liquids. Any two-phase system in which the withdrawal of a number of samples is denoted as s.
phases differ in density separates unless continu- If, instead of equal parts A and B, the proportion of
ously agitated. an active ingredient, A, in the mixture was 0.1 (10%),
 Neutral mixing occurs when neither mixing nor imposition of limits of 10% (in 997 cases out of
demixing takes place unless the system is acted 1000) requires that each sample shall contain over
upon by a system of forces. Examples are found 8000 particles. If the proportion of active constituent
in the mixing of solids and of solids with liquids is 0.01, or 1%, a figure of 90,000 particles per sample
when the concentration of the former is high. is obtained, and if the limits are reduced to þ1%, the
active constituent is 0.01, or 1% a figure of 90,000
Theoretical knowledge is, however, insufficient to particles per sample is obtained, and if the limits are
predict the performance of mixers. More commonly, reduced to 1%, the figure is 9  106.
choice is based upon broad empirical principles which The theoretical derivation of these results is based
are supported by practical tests. on component particles which vary in size, shape,
and density. This condition is not encountered in the
practical mixing of solids and, as described later, any
Mixing of Solids of these factors may prevent the formation of a ran-
dom mixture. The value of the number of particles
The mixing of all systems of matter involves a relative per sample derived in any example must therefore be
displacement of the particles, whether they are mole- raised if the limits given are to be maintained.
cules, globules, or small crystals, until a state of As already shown, a series of samples drawn from a
maximum disorder is created and a completely random random mix exhibits a standard deviation of sr. An
arrangement is achieved. index of mixing, M, suggested by Lacey[43] is given by

Unit–Validation
In 1953, Lacey[41] showed that the variation in the Eq. (32),
composition of samples drawn from a random mixture
of two materials could be expressed by Eq. (31), sr
M ¼ ð32Þ
rffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi s
pð1
pÞ
s ¼ ð31Þ where s is the standard deviation of samples drawn
n
from the mixture under examination. This approaches
where s is the standard deviation of the samples, p is unity as mixing is completed. Eq. (33) has been sug-
the proportion of one component, and n is the number gested,
of particles in the sample. The relation requires that the s0
s
two components are alike in particle size, shape, and M ¼ ð33Þ
s20
sr
density and only can be distinguished by some neutral
property, such as color. If very many samples are with-
drawn from a mixture of equal parts of two materials, where s0 is the standard deviation of samples drawn
each containing a given number of particles, the results from the unmixed materials. It is equal to p (1
p),
of analysis can be presented in the form of a frequency where p is the proportion of the component in the
curve in which the samples are normally distributed mix. It has been modified[43] to Eq. (34), using the
around the mean content of the mixture, and 99.7% variance of the samples,
of the samples will fall within the limits p ¼
s2
s2r
0.5 þ 3s. The standard deviation of the samples is M ¼ ð34Þ
inversely proportional to the square root of the s20
s2r
 3898 Unit Processes in Pharmacy: Operations

This is a fundamental equation for expressing the state relative displacement of two regions. Shear mixing
of the mixture, the index M varying from zero to one. occurs, for example, in the rearrangement of shapes
The binomial and Poisson distributions have also as the main charge falls from end to end in a double
been used to examine the state of a mixture. If the pro- cone mixer. Train[42] has stressed the importance of
portion of black particles in a random mixture of black expansion or dilation of the material so that shear
and white particles is p, the probability, P(x), of forces may be effective. A practical corollary is that
obtaining x black particles in a sample of n particles efficiency will be reduced if the machine is overfilled.
is given by Eq. (35). As long as one type of particle is not preferentially
n caught, random mixing eventually occurs in the radial
PðxÞ ¼ px ð1
pÞn
x ð35Þ plane. If, however, one component is smaller, denser,
x or has certain shape characteristics, it is preferentially
trapped and moves into the lower layers of the mixing
If p is small (<0.15) and n is large, the Poisson distri- zone until it finally concentrates as a central core run-
bution can be used, applying Eq. (36), ning the length of the mixer. Similar effects occur in
mx axial mixing, and the final shape of the segregated zone
PðxÞ ¼ e
mxl ð36Þ
formed under the influence of axial and radial move-
ment depends on the flow properties of the material.
where m ¼ np, the mean number of black particles in Similar effects have been reported with a double-cone
the samples of n particles. This relation may be used in blender.[44] Segregation also occurs with materials
an assessment of dry mixing equipment.[44] If m is dumped from the mixer.
greater than 20 and more than 10 samples are taken,
then:
Mixing rate
 About 10 of the samples have the number
pffiffiffiffi of black
particle’s outside the limits m  1:7 m, Because mixing is a process of achieving uniform ran-
 About 5% of the samples have the number domness, the rate of mixing is proportional to the
pffiffiffiffi of black amount of mixing still to be done. If, at the start a par-
particles outside the limits m  2:0 m, and,
 About 1% of the samples has the number ticle changes its path of circulation, it is most likely to
pffiffiffiffi of black find itself in a different environment. The mixing rate is
particles outside the limits m  2:6 m.
therefore high. At the end of the process, the particle is
less likely to find a different environment, and such a
Mechanism of mixing and demixing change gives no useful mixing. Fewer mixing events
take place, and the mixing rate finally reaches zero. It
The randomization of particles by relative move- can be represented for any mixing mechanism by
ment, one to another, is achieved by the following Eq. (37),
Unit–Validation

mechanisms:
dM
 Convective mixing, where groups of adjacent parti- ¼ kð1
MÞ ð37Þ
dt
cles are transferred from one location in the mass to
another. where M, the index of mixing, has already been defined.
 Diffusive mixing, where the particles are distribu- Integration of Eq. (37) gives Eq. (38).
ted over a freshly developing surface, and
 Shear mixing, where slip planes are set up within M ¼ I
e
kt ð38Þ
the mass.
The rate constant, k, depends on the physical nature of
Convective mixing predominates in machines utiliz- the materials being mixed and on the geometry and
ing a mixing element moving in a stationary container, operation of the mixer.
for example, the horizontal ribbon mixer. Groups of
adjacent particles are moved from one position to
another, steadily decreasing the scale of segregation. Mixing Machines
Diffusive mixing predominates in tumbler mixers.
The material is tumbled as it is lifted past its angle of Trough and ribbon mixers
repose. Mixing occurs when a particle changes its path
of circulation through a collision or by being trapped A simple trough mixer consists of a semicircular
in voids presented by another layer of particles. trough in which an impeller, such as a number of pad-
Shear mixing occurs when forces acting on the par- dles mounted at diverse angles on a shaft running the
ticles induce the formation of a slip place, resulting in length of the trough, rotates, lifting and distributing
Unit Processes in Pharmacy: Operations 3899

the material in an irregular manner. Convective and radial flow gives rise to axial flow by reaction at the
shear mixing occurs, as well as some fine-scale diffusive wall of the tank. Tangential flow receives no such
mixing when the impeller lifts material clear of the modification. Its predominance as laminar flow circu-
main charge. lation supports stratification at various levels. Further-
The ribbon mixer employs a ribbon-like conveying more, a vortex is created at the surface of the liquid
scroll. The helix, which may be continuous or interrupted, which may penetrate to the impeller, causing air to
is rotated in a semicircular trough and mixing again occurs be dispersed in the liquid. In general, tangential flow
through convection and shear, giving rapid coarse-scale should be minimized by moving the impeller to an
dispersion. Two ribbons set to convey material in off-center position, thus destroying the symmetry of
opposite directions are frequently fitted to the shaft. the mixer, or by modification of the flow pattern by
Although little axial mixing in the vicinity of the shaft means of baffles. Tanks with vertical agitators may
occurs, mixtures with high homogeneity can be produced be baffled by one, two, or more strips mounted verti-
by prolonged mixing, even when components differ in cally on or just away from the vessel wall. These reduce
particle size, shape, or density or tend to aggregate. but do not eliminate tangential flow, whereas little
modification of radial and axial flow occurs. Baffles
Tumbler mixers produce additional turbulence.

Tumbler mixers operate primarily by a diffusive mech- Paddle mixers

anism; their use is confined to freeflowing and granular
materials. The mild forces are employed, which pre- For a simple paddle, with upper and lower blades, suit-
clude the mixing of materials that aggregate strongly, able for mixing miscible liquids of low viscosity a tan-
allow friable materials to be handled satisfactorily. gential flow pattern predominates with zones of
The more elaborate geometrical forms are most com- turbulence to the rear of the blades (10–100 rpm).
monly used because movement of material in all The gate paddle is suitable for mixing liquids of higher
planes, which is necessary for rapid overall mixing, is viscosity and the anchor paddle with low clearance
induced. Internal baffles and lifter blades may also be between pan and blade is useful for working across a
incorporated. For example, axial movement of heat transfer surface. Stationary paddles intermeshing
material along the length of a simple drum mixer is with the moving element suppress swirling in the mixer.
slow and can be enhanced by these methods. In other examples, baffles are also necessary. Unless
paddle blades are pitched, poor axial turnover of the
liquid occurs. Paddles are therefore not suitable for
Mixing of Liquids mixtures that separate.

Miscible liquids are most commonly mixed by impellers Propeller mixers

Unit–Validation
rotating in tanks, including paddles, propellers, and
turbines. All the material should pass through the Propellers are commonly used for mixing miscible and
impeller zone at frequent intervals of time, the design immiscible liquids of low viscosity. The marine propel-
of the mixer preventing the formation of ‘‘dead’’ zones. ler is typical of the group. High speed rotation (400–
The turbulent, high velocity flow of liquid from the 1500 rpm) of the relatively small element provides high
impeller causes mixing by projecting eddies into, and shear rates in the vicinity of the impeller and a flow
entraining liquid from, the neighboring zones. The pattern with mainly axial and tangential components.
thin ribbons of one component in another rapidly They may be used in unbaffied tanks when mounted
become diffuse and finally disappear through molecular in an off-center position or inclined from the vertical.
diffusion. In large-scale operations, horizontal mounting in the
The flow pattern may be analyzed in terms of its side of the vessel is frequently used.
three components of motion:
Turbines
 Radial flow, in a direction perpendicular to the
impeller shaft. Turbine designs are intermediate between paddles and
 Longitudinal or axial flow, in a direction parallel to propellers. Turbines are effective mixers over a wide
the shaft, and viscosity range and provide a very versatile mixing
 Tangential flow, in which the liquid follows a circu- tool. The ratio of radial to tangential flow, the predom-
lar path around the shaft. inating parameters with this impeller, increases as the
operating speed increases. Pitched-blade turbines are
A satisfactory flow pattern depends on the correct sometimes used to increase axial flow. Baffles must
balance of these components. In a cylindrical tank, be used to limit swirling unless the turbine is shrouded.
 3900 Unit Processes in Pharmacy: Operations

This impeller produces a discharge with no tangential Radiation

component.
Ultraviolet light is frequently employed to reduce air-
borne microbial contamination. Surface sterilization
is usually achieved by employing a mercury vapor
STERILIZATION lamp with an emitted light of 253.7 run.
Radiation sterilization includes the use of the ioniz-
Sterilization processes do not result in a product that ing radiation of x-rays and gamma-rays. The former
can be described as absolutely sterile or non-sterile are derived from bombardment of a heavy metal target
inasmuch as the process is a statistical phenomenon. with electrons. Gamma-rays are obtained from atomic
A variety of techniques are available,[45] including heat, nucleus decay from excited to ground state.
radiation, ethylene oxide sterilization, and sterile fil- The energy evolved from radiation can be equated
tration. to photon behavior where E hn and n ¼ C/l, (E
and n are the energy and frequency of a photon,
respectively), h is Planck’s constant, and C and l are
Thermal Sterilization the speed and wavelength of light, respectively. The
energy absorbed from the radiation sources equates
The amount of heat required to sterilize depends upon to the dose.
the magnitude (T), duration (t), and amount of mois-
ture present, t 1/T. For example, heat coagulates 1 rad ¼ 100 erg/g of material absorbing
protein in the living cell. The temperature required ¼ 6:24  1013 eV=g
for this phenomenon to occur is inversely proportional
to the moisture present. ¼ 2:4  10
6 cal=gð10  10
6 J=gÞ

There are a variety of radiation sources. 60Cobalt

Dry heat decays to 59Co in the core of a nuclear reactor to emit
two photons (1.17 and 1.33 MeV) and an electron
Relatively stable substances that resist degradation at (0.31 MeV). The half-time for decay is 5.3 years.
high temperatures (>140 C) are suitable candidates 137
Cesium decays emitting one photon (0.661 MeV).
for dry heat sterilization. A 2-h exposure at 180 C or Cesium has a 33-year half-life. An electron beam can
45 min at 260 C kills spores as well as vegetative forms be accelerated to an energy equivalent of 5–10 MeV.
of micro-organisms. These exposure periods do not At energies below 5 MeV, penetration is insufficient
include the lag time from loading of the oven until ster- for sterilization. Depth of penetration can be correlated
ilization temperature is reached. The lag time depends with energy levels; for example, materials with density
on the geometry and operating features of the oven equivalent to water (r ¼ 1 g/cm3) are penetrated
Unit–Validation

and the characteristics of the load. 0.5 cm/MeV. 60Cobalt gives rise to radiation that
Both natural and forced-convection oven types can penetrates 30 cm through water. Accelerating electrons
be employed; they have been described in the section have a high dose rate and exposure is only required
on drying. The forced-convection oven offers the for seconds. 60Cobalt has a lower dose rate, and an
advantages of uniformity of heat distribution and exposure for hours is required.
reduction in lag time in comparison with the natural- Ionizing radiation arises from the photoelectric
convection system. The dry-heat method is reserved effect, the Compton effect, or ion pair production.
almost exclusively for glass or metal as other materials Gamma radiation causes local and intense damage and
char (cellulose), oxidize (rubber), or melt (plastic) at may break chemical bonds. The primary target is the
these temperatures. deoxyribonucleic acid (DNA) of the micro-organism.
In addition, free radicals may be formed, such as
Moist heat peroxides that result in intracellular and extracellular
peroxides by a chain reaction that causes damage.
Moist heat offers the advantage of greater effectiveness
at low temperatures. The thermal capacity of steam is Resistance to damage
much greater than that of hot air. Spores and vegeta-
tive forms of bacteria may be effectively destroyed in Damage depends on the amount of energy absorbed
an autoclave employing steam (121 C) under pressure relative to the number and resistance of the micro-
(15 psig) for 20 min or (27 psig at 132 C), for 3 min. organisms being irradiated. Unicellular organisms
The lag time to complete exposure of the material to have greater resistance than multicellular ones.
be sterilized is important. Gram-positive bacteria have greater resistance than
Unit Processes in Pharmacy: Operations 3901

gram-negative bacteria. Finally, bacterial spores have with propellant (88 : 12) or carbon dioxide (90 : 10).
greater resistance than vegetative forms. Viruses are Ethylene oxide polymerizes in the liquid state in
more resistant than bacteria. The energy required to 90–120 days. In this form it may plug lines or deposit
reduce the population of viruses by 90% (D value), is polymerized sludge.
0.5 Mrg i d (5 mGy). Fungi are equivalent to bacterial Ethylene oxide inactivates all micro-organisms. The
spores in their resistance. sterilizing rate depends upon its concentration, the
In order to evaluate the dose, a number of param- temperature, the duration of exposure, and the water
eters must be known. What magnitude of source content of the micro-organism. Inactivation follows
(e.g., 60Co) is available? A typical source ranges from classical first-order kinetics and is irreversible. Relative
500,000 to 2  106 Curies (Ci) where 1 Ci is humidity is synergistic, at 30–60% the micro-organism
3.7  1010 disintegrations per second. The product hydrates. The water acts as a vehicle to transport the
geometry and the speed of the conveyor carrying it gas through polyethylene and polypropylene. Poly-
to the source must be known. The dose can be evalu- styrene traps ethylene oxide and dissipates it over years
ated by a variety of dosimetric techniques. In bulk or and thus is not appropriate for ethylene oxide steriliza-
ampoules containing liquids, ferric ammonium sulfate tion. Temperatures of 40–60 C are suitable for
and ceric sulfate can be used and the absorbance heat-sensitive articles. Cycle times are longer at low tem-
change evaluated by UV spectroplidtometry; however, peratures, relative humidities, or ethylene oxide concen-
this is only accurate for 60CO and 137CS. trations. Generally, concentrations of 350–700 mg/ml
Radiochromic solids can be utilized and evaluated are employed; cycle times vary from 4–12 h.
by visible spectrophotometry. Amber and red poly- Following sterilization the load is degassed by a
methyl methacrylate are used to evaluate 0.1–1.0 Mrad dynamic process wherein filtered air is passed over
and 0.5–5.0 Mrad, respectively. Nylon film is examined the product for 12–72 h. Degassing usually takes place
for opacity following exposure and may be used to in the treatment chamber but may be moved to a sterile
evaluate exposures of 0.1–5.0 Mrad. facility. The process is monitored using Bacillus subti-
Validation requires the determination of the biobur- lis var. niger as a biological indicator, commercially
den and the D value. These represent the dose required available as spore strips (106 spores per strip). In
to achieve sterilization and the estimated dose. If low addition, the load is probed with thermocouples during
D values are obtained, the dose may be regarded validation. The gaseous mixture is sampled at different
as overkill. Bocillus pumulis exhibits inherently high points in the sterilizer for gas chromatographic analysis.
resistance to gamma-ionization radiation (D values
0.15–0.22 Mrad). The FDA prefers a 12-log reduction
in microorganisms. The dose required is approximately Sterile Filtration
2.6 Mrad.
Several filter geometries are available for sterile fil-

Unit–Validation
tration. They consist of flat membranes in a stainless
Ethylene Oxide steel press (<293 mm), pleated membranes housed in
stainless steel cartridges, and stacked plates in the form
Ethylene oxide (bp, 10.8 C) is a gaseous alkylating of flat segments of membrane filters.
agent. It alkylates proteins and ribonucleic and deoxy- Matrix filters consist of fibers with pores having a
ribonucleic acid in micro-organisms. It replaces labile depth up to 120 mm. Cellulose nitrate may be dissolved
hydrogen with hydroxyethyl groups. Ethylene oxide in a highly volatile solvent, such as amyl acetate, ether,
is utilized as a surface sterilant. Bulk crystalline materi- or dioxane. A gel-forming solvent, acetone, ethanol, or
als can occlude vegetative bacterial cells or spores with propanol, may be added. The mixture is poured on a
crystals. Consequently, ethylene oxide does not reach flat plate and placed in a controlled-temperature
them. The final step prior to sterilization is an aseptic environment to dry. Pore size is dependent on the con-
recrystallization step. centration of the gel-forming solvent. A number of
Ethylene oxide is a colorless gas with an aromatic other substances may be used as filter material, includ-
odor. The threshold limit for the odor is 700 ppm. ing cellulose, acetate and butyrate, polyamides (nylon),
The OSHA specification for worker exposure is polysulfones, fluorocarbons (Durapore membranes),
10 ppm. The toxicity of ethylene oxide is similar to that polyvinylidene difluoride (hydrophobic), or surfaces
of ammonia. It causes conjunctival and respiratory modified with organic amides (hydrophilic), acrylic
irritation, dizziness, headaches, and vomiting. It is polymers, or polyvinyl chloride. To make some mem-
known to be mutagenic and may be carcinogenic. branes hydrophilic, surfactants may be added includ-
By-products include ethylene glycol (bp, 198.9 C) ing Tween 80, Triton X-100, hydroxypropyl cellulose,
and ethylene chlorhydrin (bp, 128.4 C). Pure ethylene or glycerol. Sieve filters are made of polycarbonate
oxide is flammable and explosive. It is generally mixed (Nucleopore, 10 mm thick). Collimated uranium fission
 3902 Unit Processes in Pharmacy: Operations

products form nucleation tracks in film. Exposure to EXTRACTION AND LEACHING

chemical etching determines the pore size.
Leaching or solid–liquid extraction are terms that
Adsorption and screening describe the extraction of soluble constituents from a
solid or semisolid by means of suitable solvents. The
Most membrane filters, when wetted, have a negative process, which is used whenever tea or coffee is made,
charge. Bacteria have a similar negative charge and do is an important stage in the production of many fine
not necessarily remain on the filter. Filters with other chemicals found naturally in animal and vegetable
characteristics can be selected under these circumstances. tissue. Examples are found in the extraction of fixed
Positively charged (AMF Zeta Plus Membrane) or oils from seeds, in the preparation of alkaloids, such
protein- and peptide-adsorbing (Pall Posidyne Nylon 66) as strychnine from Nux vomica beans or quinine from
filters can be selected. Cinchona bark; and in the isolation of enzymes, such
Ionic strength, pH, pressure, and flow rate affect as rennin, and hormones, such as insulin, from animal
particle adsorption. The flow rate through a filter is sources. In the past, a wider importance attended the
expressed by Eq. (39), process because the products of simple extraction pro-
cedures, known as galenicals, formed the major part of
Ci A P the ingredients used to fulfill a doctor’s prescription.
Q ¼ ð39Þ
V Whatever the scale of the extraction, leaching is per-
formed in one of two ways. In the first, the raw
where Ci is the inherent resistance of the filter to flow (a material is placed in a vessel, forming a permeable
function of void volumes), A is the surface area, P is bed through which the solvent or menstrum percolates.
the pressure, and V is the viscosity. Filters are rated The wanted constituents are dissolved, and the solu-
according to nominal pore size and absolute pore size tion issues from the bottom of the bed. This liquid
(the largest pore in the filter); this recognizes that a is sometimes called the miscella and the exhausted
pore size distribution exists. solids, the marc. The process is called leaching by
percolation. The second process employs immersion
Filter integrity and consists of immersing the solid in the solvent and
stirring. After a suitable period of time, solid and liquid
The filter integrity can be evaluated by a number of are separated.
techniques. The destructive test involves filtering a sus-
pension of bacterial cells (Pseudomonas diminuta, Percolation
0.3  1 mm, through a 0.2 mm-filter. If 6 L of suspen-
sion containing 1  107 organisms per mL are passed Coarsely ground material is placed in the body of the
through a 1-mm filter, there should be no micro- extractor which may be jacketed for control of the
Unit–Validation

organism and an 8-log reduction would have occurred. extraction temperature. The packing must be even or
The bubble-point test assumes that pores can be char- the solvent flows preferentially through a limited vol-
acterized as capillaries. When totally wetted, all the ume of the bed and leaching is inefficient. In large
capillaries should be full of water or solution. The pore extractors, channeling is prevented or reduced by
length is generally much greater than the diameter. horizontal, perforated plates placed at intervals in the
Pressure is applied to the wetted filter. The bubble- bed; these redistribute the percolating liquid.
point pressure, P, may be described by Eq. (40), Solvent inhibition swells dried materials and reduces
the permeability of the bed. This is most marked with
4g cos yY
P ¼ ð40Þ aqueous solvents. If swelling occurs, it is necessary to
D moisten the material with water or with the solvent
before it is packed into the extractor.
where g is the surface tension (72 dynes/cm2 or 7.2 Pa),
y is the contact angle, and D is the diameter of capil-
lary. The bubble-point test is performed before and Immersion
after sterile filtration.
A specified area of filter must be soaked in a speci- In pharmaceutical processes, leaching by immersion is
fied volume of product for a designated time. The carried out in simple tanks which may be agitated by a
accelerated stability of active ingredients at 40–60 C turbine or paddle. If the solids are adequately sus-
for 60 days must be established prior to the selection pended, intimate contact between the phases promotes
of a filter for a particular purpose. The extent of dam- efficient extraction. Incomplete extraction due to chan-
age, and the nature and quantity of extractables and neling is avoided and difficulties due to swelling do not
their potency have to be evaluated. arise. Problems arise, however, in the subsequent
Unit Processes in Pharmacy: Operations 3903

separation of the phases. The materials to which leach- at which leaching proceeds. In pharmaceutical leach-
ing by immersion is applied are normally either finely ing, however, the solid matrix is usually cellular, a
divided or coarse and compressible. When agitation structure which normally offers the highest diffusional
ceases, the solids settle and the leach liquid can be resistance. The complexity of such structures does not
siphoned or pumped off by lines suitably placed in permit a strict analysis of the processes of mass trans-
the tank. The sediment, however, contains a large fer. Nevertheless, the simple diffusional concepts
volume of the leach liquid which must be recovered expressed in Fick’s law suggest that the following
by resuspending the solids in fresh solvent, allowing factors influence the leaching rate:
the solids to sediment and decanting the supernatant
liquid. Cake filtration provides an alternative method  The size distribution of the leached particles,
of separation. The leach liquid remaining in the cake  The temperature of leaching,
is displaced by passing a wash liquid. In some  The physical properties of solvent, and
cases, a filter press may be used for both extraction  The relative movement imposed upon the solids
and separation. and the liquid.

Solvent
Size and Size Distribution of the
Solid Particles
The ideal solvent is cheap, non-toxic, and non-flammable.
It is highly selective, dissolving only the wanted con-
The particle size of the solids determines the distance
stituents of the solid. It should have a low viscosity,
which solvent and solute must diffuse within the solid
allowing easy movement through a bed of solids, and,
matrix. Since this distance offers the major diffusional
if the resulting solution is to be concentrated by
resistance, its reduction by comminution raises the rate
evaporation, have a high vapor pressure. Water and
of leaching, the concentration gradient being effec-
alcohol, and mixtures of the two, are widely used.
tively increased. In addition, the inverse relationship
Both, however, are non-selective, leaching varying
between particle size and surface area requires an
proportions of gums, mucilages, and other unwanted
increase in the area of contact between the matrix
components. Most of the tinctures and liquid extracts
and the surrounding liquid. Transfer of solute at this
used in pharmacy are simple, impure extracts made
boundary is therefore facilitated. In leaching by
with, water or mixtures of water and alcohol. Acidified
immersion, a further advantage conferred by size
or alkaline mixtures of water and alcohol are used to
reduction is the ease with which finer particles are sus-
extract insulin from comminuted pancreas. A more selec-
pended. Finally, extensive cell rupture occurs during
tive extraction is given by petroleum solvents, benzene,
grinding, allowing more direct contact between solvent
and related solvents. In the preparation of many pure
and solute and more rapid dissolution and diffusion.
alkaloids, the powdered material is moistened with an

Unit–Validation
Other factors, however, operate against size
alkaline solution, packed into a bed, and leached with
reduction. Leaching by percolation demands the for-
petroleum. Subsequent purification by fractional crystal-
mation of a permeable bed. Low permeability gives
lization is facilitated by the absence of gums. Acetone
low flow rates and low extraction rates. Permeability
and chlorinated hydrocarbons also find applications in
is a complex function of both particle size and
leaching. In some cases, specific properties of the wanted
porosity, the former determining how a given void
constituents may suggest a particular solvent. Eugenol,
space is to be disposed within the bed. The disposition
for example, can be readily extracted from cloves with
of the void space consists of few channels of relatively
a solution of potassium hydroxide.
large diameter, that is, a bed of high permeability, if the
particle size is large. In leaching by immersion, the dif-
Leaching Rate ficulties of separating solid and liquid increase as the
particle size decreases.
Whatever method is adopted, leaching consists of a The opposition of the factors suggests an optimum
number of consecutive diffusional or mass transfer particle size for any particular extraction. This is
processes. The solvent first penetrates the raw material determined to some extent by the physical nature of
and dissolves the soluble elements. These diffuse in the the solids. A dense, woody structure would be
opposite direction to the surface of the solid matrix extracted as a fine powder. An example is given by
and through the liquid layers at its surface to reach the root of Ipecacuanha. A leafy structure, on the other
the bulk solution. These processes are under the hand, would be more satisfactorily leached as a coarse
influence of an overall concentration gradient, the con- powder.
centration being lowest in the bulk solution. Any of Both porosity and permeability are influenced by
these processes may be responsible for limiting the rate the particle size distribution. A high porosity is secured
 3904 Unit Processes in Pharmacy: Operations

if the distribution is narrow. Small particles may other- APPLICATIONS

wise fill the interstices created by the contact of larger
particles. After grinding it is often necessary, therefore, The processes described here are integrated in order to
to classify the product and remove undersize material. facilitate the production of pharmaceutical dosage
This material would then be bulked with the fines from forms. The following examples are intended to illus-
other batches and extracted separately. A further trate the application of the processes in the dominant
advantage arising from a narrow size distribution is pharmaceutical settings. They have been selected to
even packing and the creation of a regular system of demonstrate the broad application of unit operations
pores and waists. This promotes even movement of sol- in pharmaceutical manufacturing. Unique processes
vent and solution through the bed. are associated with each dosage form. This is no less
the case for dermatologics, intranasal and inhalation
products, and the range of alternative pharmaceuticals
Temperature
than for the examples given. Nevertheless, the majority
of processes and their underlying principles[11] are simi-
Within the limits imposed by the thermal stability of
lar from one dosage form to the next.
the wanted constituents, a high extraction temperature
appears desirable. The solubility of most materials
increases with increasing temperature, allowing higher
solute concentrations and higher concentration gradi- Parenteral Products
ents. Both this and the increased diffusivity result in
higher extraction rates. In many cases, however, mate- Parenteral products are intended for injection into a
rials are susceptible to heat degradation and cold variety of subdermal and submucosal locations.[46]
extraction must be used. In addition, the selectivity Their manufacture can be defined as a sequence of
of a solvent may be impaired at high temperatures. operations intended to be performed in certain environ-
An example of the use of moderately high tempera- ments or under specific conditions. Fig. 6 illustrates the
tures is the extraction of Rauwolfia alkaloids with boil- sequence in which these processes may be combined.
ing methanol. This flow diagram shows the relationship of the unit
operations to the underlying physical-organic chemis-
try of compounding and the subsequent processes
The Relative Movement Imposed Upon the
pertaining to packaging.
Solids and the Liquid

The major and controlling resistance to the diffusion of

the solute to the bulk solution is normally found in the Solid Dosage Forms
cell matrix. Increase in the rate of movement of the
The majority of solid dosage forms are intended for
Unit–Validation

solution past the surface does not, therefore, greatly

affect the rate of extraction. This is in marked contrast oral ingestion. The drug released from the dosage form
to the processes of dissolution and crystallization. is available at the site of absorption or action within
Nevertheless, movement is imposed upon the solvent the gastrointestinal tract.
in both general methods described above. Additional processes are required for the pro-
In the percolation of a liquid through a bed of duction of tablets beyond those described previously.
solids, mass transfer of the solute from the surfaces As these processes are not ubiquitous in pharmaceuti-
of the solid to the liquid in the interstices of the bed cal manufacturing they are dealt with only briefly here.
takes place by molecular diffusion and by natural con-
vection arising from the density changes created by dis-
solution. Although these processes are slow, they are Supply/ Aseptic Clean
Clean environment
storage area area
much faster than mass transfer in the matrix under
Formulation Compounding/filtration
the same concentration differences. Concentration
ingredients [using cleaned and sterilized
gradients in the liquid outside the particles are, there- processing equipment]
fore, very low. At any point in the bed, the introduc-
Filling/ Packaging/
tion of dilute solution from above and the loss of sealing storage
concentrated solution to below decrease the interstitial Processing
equipment
concentration by dilution or displacement. This effect
Cleaning Sterilization
can be considered simply to reduce the solute concen- Containers
tration at the junction of solid and solution, thus
imposing a favorable concentration gradient within Fig. 6 Flow diagram illustrating the various processes in
the matrix. sterile parenterals production.
Unit Processes in Pharmacy: Operations 3905

Granulation tablet manufacture considers the effect of the applied

pressure on porosity of a compressed powder.[49] Data
Following particle size reduction and blending the for- may be plotted as the negative natural logarithm of
mulation may be granulated,[47] which provides hom- porosity against applied pressure in the form of a
ogeneity of drug distribution in the blend. In Heckel plot.[50] The slope of this plot is proportional
addition, it may help flow properties and compression to the yield value (f, elastic limit) with a value of 1/3f).
characteristics of the powder. Large granules can be
prepared from primary particles by drying from a
slurry (with techniques described above) or spraying Bioprocessing
with granulating solution. Fig. 7 shows a top-spray
granulator. An alternative method (Fig. 7) employs The use of biotechnology in the manufacture of phar-
an auger to force the blend between rollers, thereby maceuticals is of increasing interest Consequently these
forming a compressed solid which disintegrates into techniques require attention in the planning of unit
large aggregates.[48] processes. Bioprocessing can be considered in terms
The steps involved in granulation begin with trans- of small-scale bioreactors, or fermenters, and the trans-
ferring powders to a mixer and blending the product. lation of such processes into large-scale economically
The granulation solution can be added and coarse viable production operations.[51,52]
milling or wet granulation begun. Finally, the product Bioprocessing is by no means a new field. The topi-
is dried and milled to an appropriate size. cality of this subject is due to the increased interest in
the use of isolated cells and microorganisms as manu-
facturing tools. It might well be argued that this tech-
Compression
nology was developed millennia ago for the purposes
of wine and beer production. More recently, the use
Compressed solids, tablets, or caplets, are prepared by
of attenuated microorganisms or isolated antigenic
placing the blend of component additives in a cylinder
materials for vaccination resulted in further develop-
or die, above a moveable piston or punch. An upper
ments. In the last decade, the interest in genetic engi-
punch is brought into the top of the piston, and pres-
neering and manipulation of the genetic code of certain
sure applied to the distal ends of the punches forces
microorganisms has produced a revolution in the
the powder into a compact (Fig. 7). The quality of
manufacturing of pharmaceuticals.
the product depends upon the cohesive forces acting
on the powder upon compression. These cohesive
forces are influenced by the selection of additives Bioreactors
in the dosage formulation. One method of evaluating
The major difference between a biotechnological
process and other pharmaceutical manufacturing

Unit–Validation
operations is the need for a bioreactor (Fig. 8). These
bioreactors may be required to produce expressed pro-
teins utilizing bacteria, yeast, insect, or mammalian

BIOREACTORS

Aerobic Anaerobic

UnMIXED MIXED

Mechanical Pneumatic

Uncoupled to Reactors coupled with

other processes separation processes

Fill Compress Eject MEMBRANES EXTRACTION EVAPORATION

Fig. 7 Sequence of events in tablet press operation. Fig. 8 Schematic of types of bioreactors.
 3906 Unit Processes in Pharmacy: Operations

A B 5. Ives, K.J. Symposium: interaction between fluids and parti-

cles. Inst. Chem. Engr. 1962, 260.
6. Maroudas, A.; Eisenklam, P. Clarification of suspensions: a
study of particle deposition in granular media. Chem. Eng.
Sci. 1965, 20, 867.
7. Salter, K.C.; Hosking, A.P. Chem. Eng. Pract. 1958, 6, 487.
8. Dickey, G.D. Filtration; Reinhold: New York, 1961.
9. Suits, C.G. The Collected Works of Irving Langmuir;
Pergamon: New York, 1961; 10, 394.
Draft 10. Hinds, W.C. Aerosols Technology, Properties, Behavior
tube and Measurement of Airborne Particles; John Wiley and
Sons: New York, 1982; 164–186.
11. Terjsen, S.G.; Cherry, C.B. Trans. Inst. Chem. Engrs. 1947,
25, 89.
12. Reist, P.C. Aerosol Science and Technology, 2nd Ed.;
McGraw-Hill, Inc.: New York, 1992.
Rotating 13. Gaden, E.L.; Humphrey, A.E. Fibrous fitters for air sterili-
stir zation. Design procedure. Ind. Eng. Chem. 1956, 48, 2172.
paddle 14. Stairmand, C.J. Trans. Inst. Chem. Engrs. 1950, 28, 130.
15. Hesketh, H.E.; EI-Shobokshy, M.S. Predicting and
Measuring Fugitive Dust; Technomic Publishing Co., Inc.:
Lancaster, PA, 1985; 1–33.
Sparger Air/CO2 16. Fuchs, N.A. The Mechanics of Aerosols; Dover Publica-
tions, Inc.: New York, 1964.
Air/CO2 17. Humphrey, A.E.; Gaden, E.L. Air sterilization by fibrous
Sterile filter media. Industr. Eng. Chem. 1955, 47, 924.
18. Cherry, G.B.; McCann, E.P.; Parker, A. The removal of
Fig. 9 Bioreactors: (A) stirred tank reactor; (B) airlift fer- bacteria from air by filtration: application to industrial-
mentor. scale fermentations. J. Appl. Chem. 1951, I, S103.
19. Scott, M.W.; Lieberman, H.A.; Chow, F.S.; Rankell, A.S.;
Johnston, G.W. Drying as a unit operation in the pharma-
ceutical industry. I. J. Pharm. Sci. 1963, 52, 284.
cells. It would be difficult to describe the various bio- 20. Scott, M.W.; Lieberman, H.A.; Chow, F.S. Pharmaceutical
reactor elements and their permutations. Some of sim- applications of the concept of equilibrium moisture con-
plest examples of bioreactors are shown in Fig. 9. tents. J. Pharm. Sci. 1963, 52, 994.
21. Newitt, D.M.; Na Nagara, P.; Papadopoulos, A.L. Trans
Inst. Chem. Engrs. 1960, 38, 273.
22. Ceaglske, N.H.; Hougen, O.A. Drying granular solids.
Indust. Eng. Chem. 1937, 29, 805.
CONCLUSIONS 23. Pearse, J.F.; Oliver, T.R.; Newitt, D.M. Trans. Inst, Chem.
Engrs. 1949, 27, 1.
Pharmaceutical manufacturing entails the combination 24. Shepherd, C.B.; Hadlock, C.; Brewer, R.C. Drying materi-
als in trays. Evaporation of surface moisture. Industr.
of a number of unit processes. The major processes Eng. Chem. 1938, 30, 388.
have been described in this article. Brief outlines of 25. Cooper, J.; Swartz, C.J.; Suydam, W. Drying of tablet gran-
ulations. J. Pharm. Sci. 1961, 50, 67.
Unit–Validation

the applications of these processes to parenteral, solid 26. Pikal, M.J.; Roy, M.L.; Shah, S. Mass and heat transfer in
dosage form, and biological materials production are vial freeze-drying of pharmaceuticals: role of the vial. J.
given. Pharm. Sci. 1984, 73, 1224–1237.
The efficiency, quality, and economy of manufactur- 27. Nail, S.N. The effect of chamber measure on heat transfer
in the freeze drying of parenteral solutions. J. Parent. Drug
ing depends upon an understanding of the individual Assoc. 1980, 34, 358–368.
operations involved in processing. In many cases, 28. Jennings, T.A. Discussion of primary drying during lyophi-
unlike in other industrial processing, safety and effi- lization. J. Parent. Sci. Techn. 1988, 42, 118–121.
29. Dushman, S.; Lafferty, J.M. Scientific Foundations of Vac-
cacy of a therapeutic agent may be affected. A guide uum Technique, 2nd Ed.; John Wiley and Sons: New York,
or introduction to the practical aspects of unit pro- 1962; 48.
cesses in pharmacy is provided here. 30. Ho, N.F.H.; Roseman, T.J. Lyophilization of pharmaceuti-
cal injections: theoretical physical model. J. Pharm. Sci.
1979, 68, 1170–1174.
31. Bond, F.C. The third theory of comminution. Trans. Amer.
Inst. Min. Metall. Engrs. 1952, 193, 484.
REFERENCES 32. Holmes, J.A. Trans. Inst. Chem. Engrs. 1957, 3S, 125.
33. Carey, W.F.; Stairmand, C.J. Recent advances in mineral
1. Ganderton, D.; Hickey, A.J. Unit process in pharmacy: fun- dressing. Inst. Min. Metall. 1953, 117.
damentals. In Encyclopedia of Pharmaceutical Tech- 34. Heywood, H. Chem. Eng. Pract. 1957, 3, 8.
nology, 1st Ed.; Swarbrick, J., Boylan, J.C., Eds.; Marcel 35. Gammage, R.B.; Glasson, D.R. Crystal changes in vateritic
Dekker, Inc.: New York, 1996; 15, 341–398. calcium carbonate during ball milling. Chem. Ind. 1963,
2. McCabe, W.L.; Smith, J.C.; Harriott, P. Unit Operations in 1466.
Chemical Engineering, 5th Ed.; Ind. McGraw-Hill, Inc.: 36. Gregg, S.J. Trans. Br. Ceram. Soc. 1955, 54, 257.
New York, 1993. 37. Cleverley, B.; Williams, P.P. Polymorphysm and changes of
3. Hickman, K.C.D. Commercial molecular distillation. Eng. infrared spectra of barbiturates during sample preparation.
Chem. 1947, 39, 686. Chem. Ind. 1959, 49.
4. Ives, K.J. Proc. Inst. Civ. Engrs. 1963, 25, 345. 38. Treasure, C.R. G. Trans. Inst. Chem. Engrs. 1965, 43, T199.
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39. Perry, R.H.; Chilton, C.H. Chemical Engineer’s Handbook, Avis, K.E. Process Engineering Applications; Interpharm Press,
5th Ed.; McGraw-Hill: New York, 1973. Inc.: Buffalo Grove, IL, 1995.
40. Dankwerts, P.V. The theory of mixtures and mixing. Banker, G.S.; Rhodes, C.T. Modem Pharmaceutics, 2nd Ed.;
Research 1953, 6, 355. Marcel Dekker, Inc.: New York, 1990.
41. Lacey, P.M.C. Trans. Inst. Chem. Engrs. 1953, 21, 53. Carey, V.P. Liquid-Vapor Phase-Change Phenomena; Hemi-
42. Train, D. Pharmaceutical aspects of mixing solids. Pharm. sphere Publishing Corporation: New York, 1992.
J. 1960, 185, 129. Carstensen, J.Y. Pharmaceutical Principles of Solid Dosage
43. Lacey, P.M.C. Developments in the theory of particle mix- Forms; Technomic Publishing Co.: Lancaster, PA, 1993.
ing. J. Appl. Chem. 1954, 4, 257. Cheremisinoff, P.N. Air/Particulate Instrumentation and Analy-
44. Adams, J.F.E.; Baker, A.G. Trans. Inst. Chem. Engrs. 1956, sis; Ann Arbor Science: Ann Arbor, 1981.
34, 91. Chulia, D.; Deleuil, M.; Pourcelot, Y. Powder Technology
45. Avis, K.E.; Akers, M.J. Sterilization. In The Theory and and Pharmaceutical Processes; Elsevier Science B.V.:
Practice of Industrial Pharmacy, 3rd Ed.; Lachman, L., Amsterdam, 1994.
Lieberman, H.A., Kanig, J.L., Eds.; Lea & Febiger: Groves, M.J. Parenteral Technology Manual, 2nd Ed.; Inter-
Philadelphia, 1986; 619–638. pharm Press: Buffalo Grove, IL, 1988.
46. Boylan, J.C.; Fites, A.L. Modern Pharmaceutics, 2nd Ed.; Groves, M.J.; Olson, W-P.; Anisfeld, M.H. Sterile Pharmaceuti-
Marcel Dekker, Inc.: New York, 1990; 491–538. cal Manufacturing; Interpharm Press: Buffalo Grove, IL,
47. Carstensen, J.T. Heterogenous systems. In Theory of Phar- 1991.
maceutical Systems; Academic Press: New York, 1973; 2, Hesketh, H.E.; EI-Shobokshy, M.S. Predicting and Measuring
223. Fugitive Dust; Technomic Publishing Company: Lancaster,
48. Doelker, E. Assessment if powder compaction. In Powder 1985.
Technology and Pharmaceutical Processes; Chulia, D., Hyman, D. Mixing and agitation. In Advances in Chemical
Deleuil, M., Pourcelot, Y., Eds.; Elsevier: Amsterdam, Engineering; Academic Press: New York, 1962; 3.
1994; 403–471. Klegerman, M.E.; Groves, M.J. Pharmaceutical Biotechnology
49. Carstensen, J.T. Pharmaceutical Principles of Solid Dos- Fundamentals and Essentials; Interpharm Press: Buffalo
age Form; Technomic Publishing Company: Lancaster, Grove, IL, 1992.
PA, 1993; 73. Lachman, L.; Lieberman, H.A.; Kanig, J.L. The Theory and
50. Heckel, R.W. Density pressure relationships in powder Practice of Industrial Pharmacy, 3rd Ed.; Lea & Febiger:
compaction. Trans. Metal. Soc. AIME 1961, 221, 671. Philadelphia, 1986.
51. Propkop, A.; Bajpai, R.K. Recombinant DNA Technology Lefebvre, A.H. Atomization and Sprays; Hemisphere Publishing
and Applications; Prokop, A., Bajpai, R.K., Ho, C., Eds.; Corporation: New York, 1989.
McGraw-Hill, Co.: New York, 1991; 415–459. Little, A.; Mitchell, K.A. Tablet Making, 2nd Ed.; The Northern
52. Hofmann, F.K. Scaleup of production and purification of Publishing Co., Ltd.: Liverpool, 1963.
cell expressed proteins. In Pharmaceutical Biotechnology, Masters, K. Spray Drying Handbook, 5th Ed.; Longman Scien-
Fundamentals and Essentials; Klegerman, M.E., Groves, tific and Technical and John Wiley and Sons, Inc.: New York,
M.J., Eds.; Interpharm Press: Buffalo Grove, IL, 1992; 1991.
138–164. Mullin, J.W. Crystallization, 3rd Ed.; Butterworth-Heinemann:
London, 1993.
Mullin, J.W. Crystallization; Butterworth: London, 1961.
Pietsch, W. Size Enlargement by Agglomeration; John Wiley and
Sons: New York, 1991.
BIBLIOGRAPHY Prokop, A.; Bajpai, R.K.; Ho, C. Recombinant DNA Technology
and Applications; McGraw-Hill, Inc.: New York, 1991.
Allen, T. Particle Size Measurement, 4th Ed.; Chapman and Tien, C. Granulator Filtration of Aerosols and Hydrosols;
Hall: New York, 1990. Butterworths: Boston, 1989.
Andrews, G.A.; Kniseley, R.M.; Wagner, H.N. Radioactive Phar- Van-Hook, A. Crystallization: Theory and Practice; A.C.S.

Unit–Validation
maceuticals; US Atomic Energy Commission: Washington, Monograph No. 152; Chapman and Hall: London, 1961.
1966. Weidenbaum, S.S. Mixing of solids. In Advances in Chemical
Ansel, H.C.; Popovich, N.G.; Allen, L. Pharmaceutical Dosage Engineering; Academic Press: New York, 1958; 2.
Forms and Drug Delivery Systems, 6th Ed.; Williams and Wert, C.A.; Thomson, R.M. Physics of Solids, 2nd Ed.;
Wilkins: Malvern, PA, 1995. McGraw-Hill, Inc.: New York, 1970.
 Vaccines and Other Immunological Products
Suresh K. Mittal
Harm HogenEsch
Department of Veterinary Pathobiology, Purdue University,
West Lafayette, Indiana, U.S.A.

Kinam Park
Departments of Pharmaceutics and Biomedical Engineering, Purdue University,
West Lafayette, Indiana, U.S.A.

INTRODUCTION Conventional Vaccines

The concept of vaccination was introduced in the Inactivated vaccines

late 18th century by Edward Jenner when he used
cowpox virus as a vaccine to protect humans against Inactivated (killed) pathogenic organisms can be used in
smallpox virus infections. This led to the develop- vaccines. This is the simplest way to produce vaccines,
ment of vaccines over the next 2 centuries to provide provided the organisms can be cultured easily. Therefore,
protection against various bacterial and viral patho- this method is often first tested to develop a potential-
gens. Undoubtedly, the effective vaccination against vaccine. As with any other technique of vaccine pro-
infectious diseases is the best method of reducing duction, this procedure is only good for some organisms.
suffering of human and animals caused by viral, There are a number of methods of inactivating patho-
bacterial, and parasitic infections. Over the last 200 genic organisms; the most common are treatment with
years, the technology of vaccine development and chemicals (formalin, formaldehyde, or propiolactate),
production has not changed significantly. This heat, or g-irradiation. Insome instances, the procedure
usually involves the use of either a killed pathogen of inactivation may enhance antigenicity of some antigens
combined with an adjuvant or a livepathogen with important in protection. Inactivated vaccines usually
reduced virulence. Apart from the tremendous suc- result in good humoral immune response after multiple
Unit–Validation

cess of killed and attenuated virus vaccines over inoculations. Because inactivated vaccines in general fail
the years, many of such vaccines do not provide to elicit effective mucosal and cell-mediated immune
satisfactory protection, and there are a number of responses, they may provide limited protection against
other disadvantages associated with these vaccines. mucosal and intracellular pathogens. Failure to inactivate
Additionally, there are important pathogens against the pathogenic organisms completely could result in dis-
which attempts to develop effective vaccines using ease instead of protection. During the 1950s, some lots
traditional approaches were unsuccessful. Various of poliovirus vaccine were not inactivated completely.[1,2]
protective viral antigens (envelope and/or capsid Now, the methods used to detect residual infectivity are
proteins or glycoproteins and other viral proteins) more stringent, therefore, inactivated vaccines are con-
and bacterial antigens (surface, internal, or fimbria sidered safe with extremely low or no chance of infection.
proteins; bacterial polysaccharides; bacterial toxins; There have been instances in which inactivated vac-
and other proteins involved in bacterial metabolism) cines led to atypical disease or enhanced disease severity.
have been identified as potential vaccine candidates. For example, in the 1960s, formalin-inactivated respira-
These protective antigens are used by various means tory syncytial virus (RSV) vaccine actually enhanced
to develop effective vaccines. The field of vaccine the disease symptoms when vaccinated children were
technology is not limited to infectious diseases but naturally exposed to RSV.[3,4] It was later discovered
has shown potential in other areas, such as cancer that a change in the antigenicity of RSV F and G glyco-
treatment, reproduction, and modulation of animal proteins[5] resulted not only in alteration in humoral
productivity. An overview of vaccine strategies is immune response but also in the Th1 and Th2 compo-
depicted in Fig. 1. nents of the CD4þ T-cell response to RSV.[6]

Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-100000445

3908 Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved.
Vaccines and Other Immunological Products 3909

Temperature-sensitive
mutants
Cold-adapted mutants
Site-directed mutants
Wild type organisms
Gene-deleted mutants
Attenuated organisms
Live Gene reassessment mutants
Bacterial expression vectors
vaccines Naturally available mutants
Viral expression vectors
Mutants generated by
adaptation to unnatural host

Vaccine Inactivated
vaccines Inactivated organisms
strategies

One or more subunits isolated Mammalian

Subunit from organisms or tissues cells
vaccines Recombinant protein produced in Bacteria
Immunogenic peptides Insect cells
Anti-idiotypic antibodies Viruses
Plants
Nucleic acid
vaccines

Fig. 1 Overview of vaccine strategies.

Live attenuated vaccines in monkeys and in monkey kidney epithelial cells.[8]

Measles virus was initially adapted to monkey kidney
Mostly attenuated organisms are being used as live cells and subsequently attenuated in duck embryo and
virus vaccines; however, in some instances, even virulent human tissue culture cell lines.[9–11]
organisms could be used, provided they are not adminis- Temperature-sensitive (ts) mutants have proven to
tered via the natural route of infection. For example, be the most useful type of mutants for a number
human adenovirus types 4 and 7 may cause acute respi- of viruses and bacteria because of their conditional-
ratory infections in humans when administered via the lethal phenotype. The (ts) mutants are produced by
oronasal route but provide protection when given orally alteration in the nucleotide sequence of a gene so that
in enteric-coated capsules.[7] the resulting protein product of the gene is unable to
There are different ways to attenuate pathogens for assume or maintain its functional configuration at the
vaccine production. Attenuation of organisms can be non-permissive (37–39
C) temperature. The protein,
achieved by growing them under abnormal conditions, however, is able to assume a functional configuration
at the permissive temperature (32–34
C), e.g., herpes-

Unit–Validation
which include cultivation in unnatural hosts or cell
lines. Some organisms are attenuated when they repli- viruses, adenoviruses, and influenza viruses. Thus, these
cate at different pH levels and/or temperatures. In mutants can replicate in mucosal sites with a lower
cells infected with multiple viruses with a segmented temperature, e.g., the nasal cavity, but are unable to
genome (e.g., influenza virus, reovirus), genome seg- cause systemic infections and disease.
ments are randomly recombined in the progeny. This A number of advantages associated with live vac-
process of recombination is known as reassortment cines are that: 1) they are cheap to produce because
and is also useful in generating attenuated viruses. the inoculum dose is relatively less; 2) they require
A natural pathogen of one host may be attenuated fewer inoculations; 3) they do not require adjuvants;
for another host, e.g., vaccinia virus worked as an atte- 4) they elicit both humoral and cell-mediated immune
nuated vaccine for small poxvirus eradication program responses; and 5) they can be inoculated by the
during the 1960s and 1970s, and turkey herpesvirus natural route of infection. Some of the disadvantages
works as an attenuated vaccine for Marek’s disease associated with live vaccines are that: 1) they are
virus (a chicken herpesvirus). In an inoculated host, usually less stable than inactivated vaccines and
the attenuated organism replicates without causing dis- may require refrigeration for storage; 2) some of
ease symptoms, thereby leading to induction of immune these vaccines under certain situations may revert
response somewhat similar to the natural infection with to virulent form in the host and thereby lead to clini-
the disease-causing organism. The Bacille Camet– cal disease; 3) they may not be recommended for
Guerin (BCG) strain of Mycobacterium tuberculosis immunosuppressed, immature, older, or pregnant
was attenuated after more than 200 passages on media hosts; 4) they may have a low level of residual viru-
containing increasing amounts of bile. The Sabin polio- lence; and 5) they may be contaminated with other
virus vaccine was attenuated by a number of passages adventitious organisms.
 3910 Vaccines and Other Immunological Products

Recombinant Vaccines Replication-defective vectors undergo an abortive

infection in an inoculated host that leads to foreign
Recombinant vector vaccines antigen expression similar to replication-competent
vectors. Replication-defective adenovirus vectors are
Viral Vectors. For the development of an effective generated by deleting the early region 1 (E1) genes.[18,19]
vaccine strategy for protection against mucosal patho- E1-deleted vectors can be grown in an E1-complementing
gens such as respiratory and enteric viruses, a vaccine- cell line, and animals immunized with such vectors elicit a
delivery system that can induce a protective mucosal protective immune response.[20] Avian poxviruses grow
immunity in the form of secretory IgA antibody, in normally in avian cells but would result in an abortive
addition to a systemic immune response, is extremely infection in mammalian hosts. Dogs and cats immunized
important. The route of vaccine delivery also plays with an avipox-rabies glycoprotein recombinant are
an important role in determining the type of resultant protected against rabies virus infection.[21]
immunity induced. A number of viruses, such as adeno-
viruses, poxviruses, herpesviruses, picornaviruses, Bacterial Vectors. Similar to viral expression vectors,
togaviruses, orthomyxoviruses, paramyxoviruses, and attenuated bacteria can be developed as vectors for
others, have demonstrated considerable potential as foreign gene expression and delivery for the purpose of
vectors for antigen delivery at mucosal surfaces.[12] multivalent vaccines. Immungenic foreign epitopes can
Immunogenic foreign epitopes can be expressed on be expressed on bacteria surfaces by modifying cell
the virus surface by modifying the viral capsid or surface proteins, fimbria, or flagella. It has been demon-
envelope protein.[13] A wide variety of foreign viral strated that M. bovis BCG strain induces both strong
antigens has been expressed in viral vectors, and humoral and cell-mediated immunity, therefore, it has
vaccination-challenge studies in experimental animals been developed as a delivery vector with the assumption
have demonstrated moderate to complete protection. that foreign proteins expressed by M. bovis in inoculated
Immunization with such vectors leads to the foreign individuals will also raise a strong protective immune
viral antigen expression similar to that of natural infec- response.[22] Because Salmonella and Vibrio colonize in
tion without causing disease. Antigenic peptides are the intestinal tract, attenuated strains of these bacteria
expressed along with major histocompatibility (MHC) were developed as vectors for mucosal delivery.[23–26]
class I and class II antigens and, thus, result in both Various bacterial vectors have been used to express
humoral and cytotoxic T-cell responses. a number of bacterial (B. pertussis, S. pneumoniae, Y.
Both adenovirus-and poxvirus-based vectors have a pestis, and L. monocytogenes), viral (herpesvirus, influ-
number of common advantages including that 1) enza virus, human immunodeficiency virus, simian
vector construction is easy; 2) relatively high levels of immunodeficiency virus, and hepatitis B virus), and
foreign protein expression are easily attained; 3) rela- parasitic (S. mansoni, and L. major) antigens.[26] Sig-
tive thermostability; 4) they have a large capacity for nificant improvements in attenuation of bacteria, and
Unit–Validation

foreign DNA insertion; 5) vector derivatives are non- the stability, localization, and expression levels of het-
pathogenic; and 6) they have a wide host range. More erologous antigens are required to market the bacterial
than one foreign antigen can be expressed in the same vector-based vaccines for use in humans or animals.
vector to provide protection against a number of To enhance foreign gene expression, ‘‘balanced
diseases by inoculation with a single vector. lethal,’’ plasmid-based expression vehicles have been
Vaccinia virus expressing rabies glycoprotein has developed.[27] A foreign antigen may form inclusion
been licensed for use to control rabies in the wildlife bodies or localize in intracellular compartment of the
population, especially raccoons, foxes, skunks, and vector thereby affecting the type, levels, and duration
coyotes.[14] Baits containing a live vaccinia-rabies gly- of immune response elicited against the antigen. The
coprotein recombinant virus vaccine are distributed Escherichia coli a-hemolysin secretion system (HSS)
in the rabies endemic area with the intention that that includes HlyB, HlyD, and TolC is involved in
rabies-susceptible wild animals that eat these baits will exporting the HlyA-fused foreign antigens to extra-
become immunized against rabies virus,[15] and this cellular compartment.[28] Using the HSS system for
approach has demonstrated satisfactory results. Vacci- attenuated Shigella dysenteriae the expression and
nia virus expressing the F and H gene of rinderpest secretion of Shiga toxin-B subunit were obtained.[29]
virus has shown potential for its use to control rinder-
pest in developing countries.[16,17] Gene-deleted vaccines
To increase the safety of viral vectors for immuno-
compromised hosts and to control their indiscriminate Many attenuated vaccines are derived after intro-
spread, replication defective viral vectors have been duction of random mutations in the genomes of
developed. These vectors can be grown to high titers various pathogens. In situations in which these
in vitro, but they are defective for in vivo replication. random mutations may be point mutations, attenuated
Vaccines and Other Immunological Products 3911

organisms may regain virulence owing to back muta- of a foreign protein can be produced in a bacterial-
tions. Because of our increased understanding of viru- expression system at a low cost. Because scale-up and
lence of various pathogens at the molecular level, one downstream processing have been well worked out
or more genes responsible for virulence has been iden- for bacterial-expression systems, they are usually first
tified in many pathogens. The genes associated with tested for subunit vaccine production. Many of
virulence may be genes involved with nucleic acid the immunogenic proteins, especially of viral origin,
replication and other non-structural and structural require secondary modifications that are important
components of the organism. This has made it possible for their antigenicity. A bacterial-expression system
to delete one or more of these genes involved in may produce proteins of altered immunogenicity
virulence—another strategy to produce safer attenu- because the bacterial system lacks many posttransla-
ated vaccines. tional processes. However, some viral glycoproteins
Pseudorabies virus has been attenuated by deleting expressed in bacteria induce protective immunity,
genes associated with viral virulence. These genes e.g., the gp 70 gene of feline leukemia virus.[37]
include the thymidine kinase gene (non-structural pro- A yeast-expressed hepatitis B virus surface antigen
tein) involved in viral DNA replication and the gC, (HbsAg)-based subunit vaccine is currently in use for
gG, and gE genes (non-essential glycoproteins) humans and has demonstrated excellent protection
involved in virus assembly.[30,31] A gene-deleted vaccine against hepatitis B virus infection.[38] This vaccine is
of pseudorabies virus has proved highly effective in an excellent example of the potential of recombinant
controlling this viral infection under field conditions. It subunit vaccines for providing protection against many
has been demonstrated that Salmonella typhimurium viral and bacterial infections.
aroA, aroB, and aroC deletion mutants fail to grow Because mammalian cells are known to process viral
in its host because of the absence of aromatic amino glycoproteins to their functional form by secondary
acid production. These genes have been targeted to modifications, they are considered one of the means
reduce the virulence of the bacterium. S. typhimurium to produce viral antigens for subunit vaccine pro-
gene-deleted mutants are capable of replication at least duction. However, the expression of such proteins in
for a short period in its host, thus raising a protective mammalian cells is usually too low. It was demon-
immune response.[32] Vaccination with gene-deleted strated that the stable expression of the transmembrane
vaccines also allows eradication of wild-type patho- anchor-deleted form of many viral glycoproteins in
gens from the population. Because antibodies against mammalian cells results in the secretion of truncated
the deleted gene product will only be developed in products in the medium in large quantities that could
infected animals, it is feasible to differentiate between be used as a subunit vaccine without further purifi-
vaccinated and naturally infected animals.[33,34] The cation. However, the removal of transmembrane
process of gene deletion not only attenuates the patho- anchor may potentially alter antigenicity of the secreted
gen but also offers a unique opportunity to insert protein. A number of viral glycoproteins that were

Unit–Validation
foreign genes for developing viral or bacterial-vectored expressed either in mammalian or in insect cells and
vaccines. secreted in form of proteins were suitable for providing
protective immune response include F and G genes of
respiratory syncytial virus,[39] the HN and F genes
Subunit vaccines of parainfluenza virus,[40] and the gD gene of bovine
herpesvirus type 1.[41]
A subunit vaccine consists of one or more immuno-
genic epitopes, proteins, or other components of a Immunogenic Antigen Production in Plants. In the
pathogenic organism. Immunogenic epitopes can be past decade, significant progress has been made in
chemically synthesized and are known as peptide the stable integration and expression of a wide variety
vaccines, e.g., peptide vaccine candidates for foot- of genes in plant cells, resulting in the creation of novel
and-mouth disease virus.[35,36] The pathogen could be plants for agricultural and industrial use. The inserted
disrupted, and one or more immunogenic proteins such genes confer resistance to insect pathogen and herbi-
as bacterial cell wall proteins; flagella or pili; and viral cides; enhanced tolerance to drought, salt, and frost;
envelope, capsid, or nucleoproteins can be purified. and improved agricultural production. Undoubtedly,
The isolation of such components in purified form improvements in plant attributes by genetic engineer-
is sometimes cumbersome and expensive. However, ing will have a great impact on agriculture production.
bacterial exotoxins can be easily purified, inactivated, However, it has been estimated that the major eco-
and used as toxoid vaccines. nomic (over 90%) gain of plant biotechnology will result
A number of expression systems including bacteria, from the use of plants as bioreactors to produce high-
yeasts, mammalian cells, insect cells, and plants are now valued products such as vaccines, industrial enzymes,
available for foreign protein expression. High amounts and other pharmaceuticals.
 3912 Vaccines and Other Immunological Products

Production of subunit vaccines in mammalian cells antigens and can be used instead of immunogens to
is usually expensive because of the low level of foreign elicit a protective immune response. Monoclonal anti-
gene expression and high processing cost. High levels idiotypic antibodies could serve as a source of antigen.
of foreign gene expression can be obtained in bacteria Anti-idiotype vaccines are useful in cases in which
and yeast, but many animal viral or mammalian pro- actual antigen is poorly immunogenic or similar to
teins expressed in these systems fail to undergo proper host antigens. Some of the pathogens against which
secondary modifications such as glycosylation, phos- anti-idiotype vaccines have been tested include Listeria
phorylation, sulfation, etc. Therefore, these recombi- monocytogenes, Streptococcus pneumoniae, hepatitis B
nant proteins may have altered antigenicity. Because virus, Semliki forest virus, and Sendai virus.[49,50] This
most mechanisms regulating secondary modifications type of vaccine is still in the developmental stage.
of proteins are present in plants, transgenic plants offer
an attractive alternative to produce functional viral,
bacterial, or parasitic proteins in large quantities at a DNA Vaccines
very low cost for subunit vaccine production.[42]
Similarly, the production of functional multimeric Immunization of mammalian hosts with a plasmid
antibody molecules in plants has made it possible to DNA containing a gene under control of a heterolo-
manufacture antibodies in bulk amounts for passive gous promoter has introduced a new approach in the
immunization.[43] area of recombinant vaccine design. The introduced
Two major strategies have been devised to produce DNA is taken up by cells, and the gene of interest is
foreign proteins in plants. These are: 1) the stable inte- expressed. The cells expressing the foreign antigen are
gration of chimeric gene into the plant genome under a recognized by the host immune system, leading to
suitable constitutive or inducible plant promoters,[44,45] humoral and cell-mediated immune responses. DNA
and 2) manipulation of plant pathogenic viruses.[46] vaccines can also be called polynucleotide vaccines or
Foreign protein expression in plants usually range nucleic acid (NA) vaccines. Such vaccines appear to
from 0.01 to 1% of the total plant protein. have the primary advantages of both attenuated and
The hepatitis B virus (HBV) surface antigen HBsAg inactivated vaccines but without their known limita-
produced in transgenic tobacco elicits an immune tions. NA vaccines elicit an immune response similar
response when injected in mice.[47] Mice fed transgenic to that obtained with live attenuated vaccines. They
potato tuber expressing B subunit of heat-labile entero- also provide safety similar to that of inactivated vac-
toxin (LT-B) of enterotoxigenic E. coli developed cines, however, without the obvious side effects of
antibodies to LT-B, particularly IgA antibodies.[44] adjuvants or animal-derived proteins.
Dalsgaard et al.[46] demonstrated that immunization The concept of NA vaccine evolved from initial stu-
of mink with the VP2 capsid protein of mink enteritis dies in experimental animals in which the inoculation
virus, expressed in cowpea after infection with modi- with naked plasmid DNA resulted in a protective
immune response.[51] After inoculation into a muscle,
Unit–Validation

fied cowpea mosaic virus, elicited a protective immune

response. Protection against challenge with virulent foot- the efficiency of cellular uptake of the naked DNA is
and-mouth disease virus (FMDV) in mice inoculated poor, and a large portion of the DNA is degraded
with the structural protein VP1 of FMDV produced in before it reaches the nucleus for transcription. To
transgenic Arabidopsis has been shown.[45] It has been increase the efficiency of DNA uptake by host cells
hypothesized that transgenic plants could serve as ‘‘edible and to reduce DNA degradation within the cell, a
vaccine,’’ thereby providing a very inexpensive mean of number of delivery systems, such as bombardment with
oral immunization.[48] gold microparticles coated with NA,[52,53] incorpor-
ation of NA into liposomes and other polycationic
Anti-idiotypic vaccines lipids,[54,55] biological erodable polymers,[56] and others,
have been developed (Fig. 2). Recently, it has been
Another approach to provide protective immune demonstrated that alginate microspheres can be used
response is the use of anti-idiotype antibodies as vac- for the encapsulation, delivery, and expression of
cines. Antibodies have unique sequences in the variable plasmid DNA[57] (Fig. 3). Inoculation of mice with
(V) region in their binding site known as ‘‘idiotypic microspheres containing both plasmid DNA and bovine
determinants’’. Some of the idiotypic determinants adenovirus type 3 (BAd3) resulted in a significant
make up the antigen-binding site (paratope) of the increase in transgene expression compared with those
antibody. The part of the antibody that binds to the inoculated with microspheres containing only the plas-
antigen is called a paratope. Antibodies to a specific mid DNA. As with other delivery systems, alginate
paratope of an idiotype mimic the epitope of immuniz- microspheres led to a stronger mucosal or systemic
ing antigen and are known as anti-idiotypic antibodies. immune response, depending on route of inoculation.[58]
Thus, anti-idiotype antibodies are mirror images of Because alginate microspheres are most likely taken up
Vaccines and Other Immunological Products 3913

Naked molecules. A variety of immunogenic antigens including

DNA HIV-1, SIV, HTLV-1, influenza virus, hepatitis B virus,
hepatitis C virus, herpesvirus, M. tuberculosis, Leishma-
nia, malaria, and many more have been expressed by
NA vaccines and have demonstrated encouraging
Liposomes Nucleic acid Gene gun results.[59–63]
immunization

Adjuvants
Biodegradable Other delivery
microspheres systems Adjuvants are compounds that, when administered in
combination with antigens, enhance the immune
Fig. 2 Methods of nucleic acid delivery.
response to those antigens. This enhanced immunogeni-
city can be measured as an increase of antigen-specific
antibody levels in serum and/or mucosal secretions, a
by macrophages and dendritic cells, it may have a posi- response against an increased number of epitopes, an
tive effect on the type of immune response elicited. increase of cell-mediated immune responses, or a combi-
A number of factors that have an impact on the level nation thereof. Adjuvants are particularly important for
and type of immune response produced by an NA vac- the induction of protective immune responses against
cine include the type of immunogen, the dosage and weak immunogens such as subunit vaccines. The
number of inoculations, the heterologous regulatory mechanisms by which adjuvants enhance the immuno-
sequences, the delivery system, the route of inoculation, genicity of antigens are not completely understood, but
and the presence or absence of immunomodulatory they include immunostimulation, altered processing of

Unit–Validation

Fig. 3 b-Galactosidase expression in tissues of mice inoculated orally with alginate microspheres containing plasmid DNA: (A)
liver; (B) intestine; and (C) spleen sections from the animal inoculated with microspheres containing bovine adenovirus type 3
(BAd3). (D) Liver; (E) intestine; and (F) spleen sections from the animal inoculated with microspheres containing LacZ plasmid
þ, BAd3. (From Ref.[57].)
 3914 Vaccines and Other Immunological Products

antigens, and sustained release of antigens (depot effect). dendritic cells. The activated dendritic cells, in turn,
A different type of immune response is obtained by activate T- and B-cells.
administration of antigens via the oral route, and this Immune responses can be divided into type 1 and
has different delivery requirements. type 2, based on the pattern of cytokine secretion
Many compounds can act as adjuvants. Their classi- and functional outcome of the immune response. Type
fication is made difficult by the variety in chemical 1 immune responses are characterized by secretion
composition and the overlapping, often poorly under- of IFN-gamma, production of IgG2a in mice, and
stood, mechanisms of action. Only aluminum adju- activation of macrophages, NK cells, and cytotoxic
vants are approved by the FDA for use in human T-cells. Type 2 responses are characterized by secretion
vaccines. Quil A is a saponin that is commonly used of IL-4, IL-5, and IL-13 and by IgG1 and IgE
as an adjuvant in veterinary vaccines and is also a production. The responses are reciprocally regulated.
component of immune-stimulating complexes (ISCOM). How the polarization of the immune response toward
These adjuvants are addressed in some detail below. type 1 or type 2 is determined is not exactly under-
A detailed discussion of other types of adjuvants stood. IL-12 is an important factor that drives the
can be found in recent books[64–66] and reviews[67] on this type 1 response, and IL-4 is implicated in the type 2
subject. response. Microbial products such as LPS and bac-
terial DNA stimulate the secretion of IL-12 by
Immunostimulation dendritic cells and preferentially induce type 1 immune
responses.
The immune system can be divided into the adaptive It is likely that the primary mechanism by which
immune system, comprising of B and T lymphocytes, adjuvants stimulate the immune response is by direct
and the innate immune system, which includes neutro- or indirect signaling through pattern-recognition and
phils, macrophages, dendritic cells, and soluble factors danger signal receptors. Very strong adjuvants are
such as the complement system. The innate immune often composed of or include microbial components
system plays a critical role in the activation of the such as LPS and mycobacteria or derivatives thereof.
adaptive immune system. Dendritic cells are antigen- These type of adjuvants bind to pattern-recognition
presenting cells that integrate the signals from the receptors to stimulate IL-12 production and a type 1
innate immune system and activate T-cells and pos- immune response. Coadministration of cytokines can
sibly B-cells. T-cells have antigen-specific receptors that directly activate and influence dendritic cells and the
recognize peptides displayed by MHC I molecules outcome of the immune response. This was clearly
(CD8þ cytotoxic T-cells) and MHC II molecules demonstrated with an experimental Leishmania vaccine
(CD4þ T helper cells). Engagement of the antigen- using IL-12 as an adjuvant. Immunization of genetically
specific T-cell receptor is not sufficient, and T-cells also susceptible BALB/c mice with a Leishmania antigen did
need to receive costimulatory signals delivered via not result in protection, but when IL-12 was injected
Unit–Validation

CD28 and CD40-ligand. Dendritic cells express both with the antigen, the mice became markedly resistant
MHC I and MHC II and, on activation, increase the to infection. The effect of IL-12 correlated with increased
expression of the costimulatory molecules CD80 and IFN-g and decreased IL-4 secretion by antigen-specific
CD86 (ligands for CD28) and CD40. The signals that T-cells in vitro.
activate dendritic cells include microbial molecules.
The innate immune system is equipped with receptors
(called pattern-recognition receptors) that can recog- Altered Processing of Antigens
nize molecules that are expressed by pathogens, but
not by mammalian cells, and alert the innate immune Most T-cells that carry the a–b-T-cell receptor do not
system on infection. These molecules, pathogen- recognize and react with intact proteins. Instead, the
associated molecular patterns, include lipopolysac- T-cells recognize small peptides that are derived from
charides (LPS), mannose, and bacterial DNA with proteins and that are linked to MHC I and MHC II
unmethylated CpG motifs. In addition, dendritic cells molecules. The MHC I-linked peptides are generated
are stimulated by host cell components that are in the cytoplasm (endogenous pathway) and recog-
expressed and/or released by cells when they undergo nized by CD8þ T-cells. Proteins in the cytoplasm are
stress and pathologic cell death (necrosis). The identity degraded by a complex of proteolytic enzymes, the
of these components, called danger signals, is uncertain proteasome, and the peptides are transported into the
but may include heat shock proteins. The microbial rough endoplasmic reticulum where they associate
molecules and danger signals can directly activate with MHC I molecules. Peptide binding stabilizes the
dendritic cells, or they can activate other components MHC I molecules, and the complexes are transported
of the innate immune system resulting in the secre- to the cell surface. In contrast, proteins that enter cells
tion of cytokines and other mediators that activate by endocytosis are partially degraded into peptides in
Vaccines and Other Immunological Products 3915

endosomal vesicles. The peptides bind MHC II mole- resulting in precipitation. The alum-precipitated adju-
cules that have been transported from the endoplasmic vants resemble aluminum phosphate in their chemical
reticulum to the endosomes. The MHC II–peptide and physical properties.[71] The surface charge and
complexes are then displayed on the cell surface and morphology of the aluminum adjuvants affect their
are available for recognition by CD4þ T-cells. adsorptive capacity. The rate and degree of adsorption
Vaccines that contain single proteins or inactivated are further dependent on the pH, ionic strength of the
pathogens can readily activate CD4þ T-cells because antigen solution, and isoelectric point of the antigen.
the antigens are endocytosed and processed by MHC Aluminum adjuvants are universally used in
II–positive antigen-presenting cells. Activation of the diptheria–tetanus–pertussis (DTP) vaccines and in
CD4þ T-cells can result in a type 1 or a type 2 immune most hepatitis B vaccines and have an excellent safety
response, depending on the type of adjuvant included. record. They are not ideal adjuvants, however, because
However, such vaccines usually do not activate CD8þ the enhancement of the immune response is relatively
cytotoxic T-cells because activation of CD8þ T-cells weak, they are not effective with all antigens, and, most
requires processing of antigen via the endogenous important, they only enhance the humoral (type 2)
pathway. Certain adjuvant formulations such as lipo- immune response and have little effect on the cell-
somes, the saponin QS-21, and poly-(lactic-co-glycolic mediated (type 1) immune response.
acid) (PLGA) are able to induce cytotoxic T-cell The mechanism by which aluminum enhances the
responses to protein antigens.[68] These adjuvants immune response is not clear. Early studies suggested
appear to target some of the injected antigens into that aluminum adjuvants slowly release the adsorbed
the cytosol of antigen-presenting cells for processing antigen over time (depot effect). However, recent
via the endogenous pathway. The mechanism by which experiments demonstrated that antigens are rapidly
this occurs is not known. desorbed after injection in animals. Moreover, alumi-
num phosphate enhanced the immune response to
Sustained release of antigens DNA-encoded antigen after DNA immunization,
clearly indicating that adsorption may not be critical
The slow and continued release of antigens has been to the adjuvant effect of aluminum compounds. These
postulated to induce a strong immune response through data indicate that aluminum enhances the immune
continued activation of the immune system. This may response via other mechanisms. A satisfactory expla-
contribute to the adjuvant effect of aluminum-based nation of the adjuvant effect of aluminum also needs
adjuvants and mineral oils. Newer technologies may to take into account its selective mode of enhancing
allow for the design of vaccines that release antigens the immune response, i.e., a predominant type 2
from a depot at certain time intervals after a single immune response. Aluminum adjuvants induced differ-
injection. One example is the use of poly PLGA micro- entiation toward type 2 immune responses, even in the
spheres for encapsulation of antigens. By varying the absence of IL-4 or IL-13. Aluminum stimulated a type 1

Unit–Validation
polymer composition and size of the microspheres, and type 2 immune response in genetically engineered
the release of antigen can be varied. Pulsatile release mice with a defective IL-4 and IL-13 response, suggest-
of antigen can be attained by combining multiple ing that aluminum-induced IL-4 and/or IL-13 secretion
variations of PLGA microspheres in a single dose of suppresse the type 1 response but are dispensable for a
the vaccine.[69] Relatively little is known about the type 2 response in intact animals.[72] The lack of a type 1
desired pattern of antigen release to obtain a maximal immune response is a drawback for the use of aluminum
response. It was recently suggested that continued in vaccines for intracellular pathogens and tumors.
release of antigen is not desirable for the induction of A recent study demonstrated that aluminum adjuvant
strong memory cell responses. Mathematical models with adsorbed IL-12 induces a strong type 1 response,
may help design appropriate strategies for the release indicating that it is possible to overcome the aluminum-
of antigens from depots after a single injection.[70] induced suppression of type 1 responses.[73]

Saponins
Aluminum
The saponins of the bark of the Quillaja saponaria
Aluminum adjuvants in human vaccines are either Molina tree have long been known to have immuno-
aluminum hydroxyphosphate (commonly referred to stimulatory activity. A partially purified fraction, Quil
as aluminum phosphate) or aluminum oxyhydroxide A, has reduced toxicity and more potent adjuvant
(aluminum hydroxide).[71] Aluminum-based vaccines are activity and is used in veterinary vaccines. Quil A can
prepared by adsorption of antigen to commercial alumi- be further fractionated into fractions that have differ-
num hydroxide or aluminum phosphate gels or by mix- ent degrees of toxicity. QS-21 is a less toxic component
ing antigen with alum (potassium aluminum sulfate), with strong adjuvant activity. Saponins probably act
 3916 Vaccines and Other Immunological Products

by direct stimulation of the immune system.[74] They stimulate a response in the Peyer’s patches, a major
stimulate both the humoral (primarily IgG2a anti- inductive site for mucosal responses.[76] For many
bodies in the mouse) and cell-mediated immune other antigens, however, the usefulness of parenteral
responses. QS-21 causes protein antigens to be pro- vaccination is limited by the insufficient induction of
cessed and presented via the MHC I pathway, resulting mucosal immune responses.
in cytotoxic T-cell responses. Cytokine analysis indi- Parenteral vaccination is difficult for those living in
cates that QS-21 stimulates type 1 cytokine production. the developing countries where medical care is not
Immune-stimulating complexes (ISCOMs) are 30– well-established. Vaccination of a large number of
40 nm particles consisting of Quil A, cholesterol, subjects using hypodermic needles, which is a highly
antigen, and phospolipids.[74] They are used in a commer- labor-intensive procedure requiring healthcare person-
cial vaccine for equine influenza. ISCOM-adjuvanted nel, is not practical. The problem becomes even more
vaccines stimulate a strong humoral and cell-mediated significant for vaccination of millions of animals. For
immune response caused by the immunostimulatory example, vaccination for routine control of Newcastle
actions of Quil A and targeting of the particles to macro- disease in chickens by intramuscular injection requir-
phages. As with Quil A, ISCOMs target antigens for ing individual handling of the birds is not practical.[77]
processing via the MHC I pathway, resulting in induction Recent advances in needleless injectable systems have
of cytotoxic T-cell responses. made the parenteral vaccination easier, but it still
requires individual handling. Examples of needleless
injection systems are PowderJectÕ, Medi-JectorÕ,
Delivery of Vaccines BiojectorÕ, VitajetÕ, Bio-SetÕ, and IntrajectÕ. They
all use high pressure released in a very short period
Parenteral vs. mucosal route to deliver drugs through the skin. A jet-immunization
technique was used for intraoral administration of
The success of vaccination depends primarily on the DNA in the cheek, resulting in high IgA mucosal
method of presenting the antigen to the host immune responses.[78] The intraoral jet-injection technique for
system. Antigens have usually been delivered by par- DNA vaccine delivery has the advantages of being a
enteral (such as intravenous, intramuscular, intraperito- simple and rapid way to administer the DNA in solution
neal, intradermal, and subcutaneous) administration, and to provoke specific mucosal IgA after administra-
but recent studies have shown that other routes of deliv- tion in the mucosal-associated lymphoid tissue.
ery such as intranasal, oral, and transdermal delivery The results of parenteral vaccination depend on the
have also been effective. In some cases, vaccination route of administration. For plasmid DNA vaccines,
through mucosal routes resulted in better responses in the highest levels of antibodies were induced by intra-
IgA production. Because non-parenteral vaccine delivery muscular and intravenous injections, although signifi-
presents many obvious advantages, numerous attempts cant titers were also obtained with sublingual and
have been made on the development of non-parenteral
Unit–Validation

intradermal delivery. Delivery to the skin by the gene

delivery of vaccines. gun induced exclusively IgG1 antibodies (Th2-like) at
4 weeks and only very low IgG2a levels at later times.
Parenteral Route. Parenteral vaccination remains the Other routes, such as intraperitoneal, intraperineal,
immunization method of choice for most antigens subcutaneous, intranasal inhalation, intranasal instil-
because it provides more effective immune response lation, intrarectal, intravaginal, ocular, and oral, did
than do any other routes of vaccination in most cases. not result in significant immune responses.[79]
Every years millions of people receive inactivated influ- Dual-chamber syringe. For delivery of two estab-
enza vaccine by parenteral administration. Subcuta- lished vaccines (e.g., polyribosyl ribitol phosphate
neous vaccination with inactivated influenza vaccine conjugated to tetanus toxoid and diphtheria–tetanus–
is known to induce simultaneous immune responses whole cell pertussis and inactivated poliovirus vaccine)
in the blood and upper respiratory tract of subjects. at the same time, a dual-chamber syringe delivery sys-
The immune response, i.e., the increase in the number tem can be used. The proximal chamber may contain a
of influenza virus-specific antibody-secreting cells in vaccine in the freeze-dried solid state, and the distal
peripheral blood and tonsils, increased rapidly to reach chamber contains a vaccine in the liquid formulation
a peak within 1 week after vaccination.[75] Parenteral that allows reconstitution of the vaccine in the proxi-
vaccination of a DNA vaccine encoding glycoprotein mal chamber. The immune response by the dual-
D of herpes simplex virus type 2 resulted in systemic chamber delivery of vaccination was equivalent to that
cellular and humoral responses. The mucosal humoral by the separate-injection method of vaccination. The
responses generated by intramuscular and intradermal dual-chamber syringe can be used for safe and effective
vaccination were comparable with those obtained by delivery of two different vaccines that are not yet avail-
mucosal vaccination. The DNA vaccine was able to able as a single formulation for pediatric applications.[80]
Vaccines and Other Immunological Products 3917

Intranasal Intratracheal Oral Rectal

application application application application

Nasal-Associated Bronchi-Associated Gut-Associated Large intestinal

lymphoid tissue lymphoid tissue lymphoid tissue follicles

Central Hilar Mesenteric Rectal

lymph nodes lymph nodes lymph nodes lymph nodes

Common Mucosal Immune System

(Blood and lymph tissue)

IgA in exocrine IgA in upper IgA in IgA in

glands (mammary, respiratory gastrointestinal genitourinary
salivary & lacrimal) tract tract tract

Fig. 4 Mucosal immunization and production of IgA antibodies in various mucosal surfaces via the common mucosal-simmu-
nization system. Nasal and rectal vaccinations usually result in IgA production in upper respiratory tract and genitourinary tract,
respectively, whereas effector sites by oral vaccination are expected to include many mucosal surfaces.

The primary advantage of the dual-chamber syringe stimulate IgA precursor cells that may then migrate
is that it reduces the cost of vaccine delivery and, at to other mucosal surfaces to elicit immune reaction
the same time, increases the vaccine acceptability and in other mucosal tissues. It is known that the mucosal
coverage rate of vaccines.[81] immune system produces 70% of the body’s anti-
bodies.[85] Fig. 4 shows a schematic description of the
Mucosal Route. Vaccination through mucosal routes common mucosal-immunization system. Mucosal
provides new avenues of vaccination with a unique delivery of numerous antigens by a variety of routes
advantage of mucosal immunity, that may not be (oral, nasal, tracheal, and rectal) has been shown to eli-
obtained, through parenteral vaccination. Mucosal cit immunity at mucosal surfaces mediated by

Unit–Validation
immunization presents a realistic alternative to par- secretory IgA. The presence of MALT indicates that
enteral administration for inducing protective immune mucosal vaccination at a certain site in the body can
responses. Vaccination by mucosal route provides a be achieved by mucosal immunization at the distal site
number of advantages over parenteral vaccination. of the body. Although the mucosal and systemic
First, mucosal vaccination does not involve hypoder- humoral immune systems function essentially inde-
mic needles, which are not user-friendly. Second, the pendent of each other, an antigen administered by
total surface area of the mucosal surfaces in the gastro- one route can modify responsiveness to subsequent
intestinal, respiratory, and urogenital tracts where immunization by an alternate route.[86]
many infectious pathogens come into contact with Oral vaccination of the various mucosal routes,
the host is huge. Thus, preventing infections at the oral vaccination is the most preferable mode of vacci-
mucosal surface provides an immunological first line nation because of its ease of use and low cost of manu-
of defense against diseases.[82] This makes priming of facturing.[87] Furthermore, the gastrointestinal (GI)
the mucosal-associated lymphoid tissue (MALT) by tract provides the largest component of the mucosal
vaccination most desirable. Parenteral vaccination immune system that has been well-characterized. Oral
alone is quite often insufficient in inducing mucosal administration of vaccines has high acceptability, by
immune responses, because stimulation of the MALT avoidance of injection, to individuals of all ages.
usually requires direct contact between the immunogen Fig. 5 shows the current understanding of oral vacci-
and the mucosal surface.[83] The mucosal tissues are nation. After oral vaccination, an antigen, which is
protected by interconnected local immune system, typically loaded in microspheres, is taken up by M-cells
which is essentially separated from systemic immun- in the Peyer’s patch of the gut-associated lymphoid
ity.[84] In a common mucosal-defense system, an anti- tissue. The antigen is then passed to the macrophages
gen interacting with localized lymphoid tissue can and B-cells (B). These cells in turn present the antigen
 3918 Vaccines and Other Immunological Products

Vaccine are not effective. Thus, prevention of the antigen

Nasal degradation is the first step toward successful oral vac-
Pulmonary cination. Adding protease inhibitors before oral vacci-
Secretory IgA
(sIgA) nation may induce complete immunity, but this
Oral approach is not practical. There are many different
Rectal enzymes that may not be inhibited by a particular pro-
Epithelial cell tease inhibitor, and, more important the action of pro-
Microparticles
tease inhibitors may not occur at the same time that
the antigens are present in the GI tract. Second, the
Secretory Dimeric IgA
component (or polymeric IgA) systemic uptake of antigens from the GI tract is very
M cell
poor. Even after oral intake of gram quantities of anti-
gen, only a nanogram range of antigenic material was
Macrophage found to pass the intestinal barrier.[90] It is also poss-
B ible that for certain antigens, oral vaccination may sim-
Plasma cell ply be less effective than parenteral vaccination in
Th induction of systemic immunity.[91] The protection
B resulting from oral vaccination is known to last for a
MLN TD Blood
Th relatively short period, ranging from a few months to
Mucosal membrane
1 year. To obtain the desirable immunity equivalent
Fig. 5 Mucosal immunization by oral vaccination. to systemic immunization, oral vaccination requires
much higher and more frequent oral doses. The use of
highly effective adjuvants in oral vaccine formulation
to T helper lymphocytes. These cells migrate into the may result in strong and long-lasting immunity in
blood via the mesenteric lymph nodes (MLN) and mucosal tissues.
the thoracic duct (TD). These cells subsequently loca- The issues of degradation of antigens in the GI tract
lize in the effector sites, i.e., mucosal membranes of and the poor systemic uptake of antigens from the GI
the GI tract, upper respiratory tract, genitourinary tract have led to encapsulation of antigens in micropar-
tract, and glandular tissue. At the effector sites, the ticles (also called microcapsules or microspheres). Anti-
migrating B-cells develop into plasma cells that pro- gens that are encapsulated in microparticles are
duce IgA antibodies. Polymeric IgA is then released protected from degradation, and the microparticulate
as secretory IgA (sIgA) through epithelial cells. nature allows better uptake by the M-cells in the Peyer’s
The maximal intestinal immunization can be patches. A large number of studies have shown that anti-
achieved by intra-Peyer’s patch immunization, and gens orally delivered in microparticles resulted in good
thus this method can be used to screen oral vaccine mucosal immunity. It is noted here that virus itself can
candidate antigens without the added complication of
Unit–Validation

be regarded as a particulate vaccine-delivery system.

simultaneously testing oral-delivery systems.[88] Many viruses are highly effective in inducing immuni-
Immunization of subjects against Helicobacter pylori zation after oral vaccination. Norwalk virus, which is a
by intra-Peyer’s patch resulted in an 84–91% reduction major cause of epidemic gastroenteritis, was immuno-
in H. pylori infection compared with unimmunized genic in healthy human adults even when administered
controls. The therapeutic efficacy of the recombinant without adjuvants.[92] Influenza virus can also elicit
H. pylori urease vaccine in mice was shown to be com- immune response after oral administration. Successful
parable with that achieved with the combined anti- oral vaccination relies on targeting of microparticles to
biotic/antacid treatment in humans. The oral the Peyer’s patches. It is known that the surface chemis-
vaccination is preferred to conventional treatment of try of microparticles affects the targeting to and uptake
ulcers because it is a very simple and quick procedure by M-cells in the Peyer’s patches.[93] The exact relation-
compared with long-term conventional treatment. In ships between the surface chemistry and the uptake by
addition, vaccines use the defense mechanisms of the Peyer’s patches, however, have not been fully under-
body to establish long-lasting immunity.[89] stood. Development of better oral vaccines requires
One of the limitations of oral vaccination is that it understanding of such relationships.
does not always induce sufficient immunity. There Intranasal vaccination route has received growing
are a few good reasons for this. First, the GI tract is interest for non-invasive immunization. Intranasal
designed to digest proteins by acidic and enzymatic immunization has been quite effective for various vac-
degradation for absorption. Because most antigens cine-delivery systems. Both solution and microsphere for-
are proteins in nature, they may be degraded by mulations tend to show good immune responses after
enzymes in the GI tract as well as by acids in the sto- intranasal administration. Immunization of mice with
mach. This is why soluble antigens administered orally tetanus toxoid, in solution and microsphere-encapsulated
Vaccines and Other Immunological Products 3919

formulations, resulted in high levels of specific IgG and diameter is used for the maximum alveolar (deep lung)
IgA antibodies.[94] Nasal vaccine delivery is known to deposition.[101]
be superior to oral delivery in inducing specific IgA and Direct gene transfer into the respiratory system can
IgG antibody responses in the upper respiratory tract.[95] be carried out for either therapeutic or immunization
Nasal immunization is also known to be preferable to the purposes. Cells in the lung can take up and express
oral route for distant mucosal vaccination that might be plasmid DNA whether it is administered in naked form
used to prevent adhesion of pathogens to the urogenital or formulated with cationic liposomes. For a given
tract.[95] It is interesting to note that the volume of dose of DNA, the results can be improved when the
the nasally instilled vaccine is important.[94] The larger- DNA is mixed with the minimum amount of lipid that
volume (e.g., 50 mL) of microsphere suspension resulted can complex it completely.[103] Such a complex forma-
in the higher percentage of particles entering the lungs tion can be considered a formation of microparticles
than did the lower, volume (e.g., 10 mL) instillation. that can enhance cellular uptake and subsequent
It is generally believed that microspheres that immune responses.
adhere to the nasal mucus elicit better immune
response, and for this reason, many microspheres made Parenteral and Mucosal Combination Vaccination. The
of mucoadhesive polymers, such as chitosan, have been combination of mucosal and systemic immunization
used extensively in the preparation of nasal vaccine routes (e.g., parenteral immunization followed by oral
formulations. immunization or vice versa) generally induces mucosal
Transdermal vaccination or transcutaneous immu- immune responses that are superior to immunization
nization, is attractive, because it does not require spe- by either route alone.[91] Pigs showed some protec-
cially trained personnel necessary for needle injections. tion after intramuscular inoculation with formalin-
Topical application of antigens to intact skin has shown inactivated M. hyopneumoniae vaccine in incomplete
promising results for the administration of DNA-based Freund’s adjuvant and a booster inoculation with the
vaccines. Noninvasive gene delivery by pipetting adeno- same vaccine in microspheres onto the mucosal surface
virus- or liposome-complexed plasmid DNA onto the of Peyer’s patches by a surgical operation.[104]
outer layer of skin was able to achieve localized trans-
gene expression within a restricted subset of skin in mice.
It also elicited an immune response against the protein Antigen delivery systems
encoded by the DNA.[96]
For improved results, transdermal electroporation The primary goal of antigen-delivery systems is to
was also tried to explore the feasibility of non- maintain a stable dosage form during storage and,
adjuvant, needle-free skin immunization.[97] The trans- when administered to present antigens to elicit a vigor-
dermal electroporation route elicited higher responses ous immune response in vivo. It is necessary to develop
to a myristylated peptide than did intradermal immu- vaccine formulations that would preserve the antigen
and deliver it to a specific target organ over a desired

Unit–Validation
nization. For diphtheria toxoid, however, the result
was the opposite. It appears that transdermal electro- period. Continuous release or multiple pulsatile release
poration is a promising technique for non-adjuvant skin during the desired period would eliminate the incon-
immunization, especially with low-molecular-weight, venience of multiple vaccine administration for obtaining
weakly immunogenic antigens. Topical application of satisfactory immune responses. The antigen-delivery
antigen and cholera toxin or bacterial exotoxin to the system plays one of the most crucial roles in the outcome
skin surface resulted in detectable antigen-specific IgG of the immunization. The way that antigens are delivered
in plasma and mucosal secretions.[98,99] It appears that affects the immune response significantly. Currently,
transcutaneous immunization can induce potent, protec- antigen-delivery systems are classified into two systems:
tive immune responses to both systemic and mucosal live attenuated microorganisms and non-living micro-
challenge.[100] particulate systems.
Pulmonary vaccination is especially useful in mass
vaccination campaigns. A conventional method of Live Attenuated Organisms. Live attenuated bacteria
pulmonary delivery of drugs using metered-dose, and viruses have been used not only as vaccines but
propellant-driven, small-particle aerosols was used to also as a delivery system that elicits humoral, mucosal,
deliver killed whole bacterium vaccines. The results and cellular immune responses against exogenous
showed good stimulation of mucosal immunity against antigens. Since the success with live attenuated oral
respiratory infections in animals[101] Recent advances vaccines against tuberculosis and polio more than
in powder inhaler devices have made it possible 3 decades ago, a number of live attenuated micro-
to deliver vaccines via the pulmonary route using organisms have been used as antigen-delivery systems.
dry powder inhalation technologies[102] Dry pow- Live vaccines are relatively easy and cheap to manu-
der vaccine in the size range from 1 to 5 mm in facture, because they do not require purification of
 3920 Vaccines and Other Immunological Products

antigens or formulation with adjuvants.[82] Attenuated Non-living Microparticulate Delivery Systems. Non-
strains of microorganisms can be formed sponta- living immunogens generally result in immune responses
neously or induced by heat, chemical, or UV mutagen- of lesser magnitude and of shorter duration than do
esis. Another advantage of the attenuated live vaccines those by living immunogens.[82] Nonliving immunogens
is that they can be administered by the natural route of are usually made of microparticulate forms to protect
infection. Recently, pathogenic microorganisms have antigens and to improve cellular uptake. Nonliving
been attenuated by genetic engineering, i.e., mutating microparticulates that can be used as antigen-delivery
specific genes or removing some toxic genes. Because systems include polymeric microparticles, liposomes,
much of the infection occurs through the mucosal sur- virus-like particles, neosomes, and cross-linked protein
faces, live attenuated vaccines are best suited for pro- crystals. The definition of microparticles should be
tection against pathogens that access the body broad enough to include all other forms, such as protein
through the mucosal surfaces. Live attenuated oral aggregates. The size of microparticles used in the vaccine
vaccines are expected to provide the most convenient area is usually less than 50 mm.[110] It is common,
and effective means of vaccinating against enteric dis- however, to call any particles less than a few hundred
ease.[105] Orally administered attenuated Salmonella micrometers microparticles. For this reason, it is impor-
are known to interact with the MALT.[82] Other exam- tant to specify the average size of microparticles for
ples of live attenuated microorganism vaccines particular applications, because the size of microparticles
are BCG (bacilli Calmette–Guérin), adenovirus, and often affect the outcome.
poliovirus. Polymeric microparticles and liposomes have been
Some viruses and bacteria are inherently quite used extensively as controlled-release dosage forms
stable. For example, polio virus can be formulated for many drugs including antigens. They have been
as a frozen liquid. A live poliovirus vector expressing quite useful in oral delivery of antigens because encap-
a foreign antigen generates both antibody and cyto- sulation in microparticles can protect antigens from
toxic T-lymphocyte responses in mice.[106] Most live acidic and enzymatic degradation in the GI tract,
bacteria and viruses, however, are usually stored as and thus serve as a stable vaccine vehicle with extended
powders after freeze-drying or lyophilization. Preser- shelf life. Delivery of antigens by microparticulate-
ving the live state through freeze-drying often requires delivery systems has the potential benefits of reducing
the presence of a stabilizer, which is selected primarily the number of inoculations, enhancing the immune
through trial and error. The most widely used non- response via both parenteral and oral vaccination
specific stabilizers are sugars, amino acids, polyols, routes, and reducing the total antigen dose required
and neutral salts which are known to act as bound to achieve immune protection.[111] Microparticulate
water substitutes for maintaining the conformational vaccine-delivery systems show improved immune
integrity of proteins. An example of lyophilized vac- responses because of the protection of the loaded anti-
cine products is S. typhi bacteria lyophilized to a gens from degradation and the slow release of the
Unit–Validation

powder that is encapsulated into gelatin for oral antigens. For this reason, microparticulate-delivery
administration.[107] systems are often considered adjuvants.[66]
One of the drawbacks of using live microorganisms Polymer microparticles, a large number of poly-
is that attenuated pathogens may invoke the very dis- mers, such as poly(methyl methacrylate), poly(butyl
ease they are designed to prevent if they are insuffi- cyanoacrylate), poly(lactide-co-glycolide), polyarcyl-
ciently attenuated. Even if they are sufficiently starch, dextran, albumin, and alginic acid, have been
attenuated, they still may cause severe infections in used for making microparticles for vaccine delivery.
immunocompromised individuals. In addition, they All the polymers that have been used for controlled
always have a potential to revert to full virulence if drug delivery can be used for vaccine delivery.[112]
lesions causing attenuation are not fully character- Preparation of microparticles from water-insoluble
ized.[82] If pathogens are overattenuated, they fail to polymers [e.g., poly(methyl methacrylate), poly(butyl
trigger an appropriate immune response. Thus, it is cyanoacrylate), and poly(lactide-co-glycolide)] requires
highly important to attain the right balance between use of organic solvents or high temperature, both of
minimal virulence and maximal immunogenicity. This which may not be good for maintaining tertiary struc-
balance can be achieved in a normal population but tures of antigens. Preparation from water-soluble poly-
may not be the same in a population with even minor mers frequently requires cross-linking reaction to make
defects in immune competence.[108] Another aspect to the polymers remain insoluble. It is possible that cross-
notice in using live vaccines is that the distribution linking agents cross-link not only polymer chains but
of live vaccines requires a cold chain that may not also antigen molecules. Absorption of water into
be readily accessible in many developing countries, hydrophilic polymers results in swelling of the net-
and this may offset advantages of using live-vectored work, i.e., formation of hydrogels, or aquagels. Prep-
vaccines.[109] aration of microparticles from hydrophilic polymers
Vaccines and Other Immunological Products 3921

is preferred because it does not require organic solvents induce strong antibody, helper T-cell, and cytotoxic
or high temperature. Polymers that have been used in T-lymphocyte responses.[114] Parenteral administration
the immunization vary depending on the route of of recombinant VLPs of papillomavirus induced VLP-
administration. specific humoral and cellular immune responses.[115]
For parenteral vaccination, biodegradable poly- Immunization of VLPs without adjuvant via mucosal
meric microparticles made of poly(lactide-co-glycolide) route is also known to elicit specific antibody at muco-
are commonly used as vaccine carriers. Poly(lactide-co- sal surfaces and also systemic VLP epitope-specific
glycolide) has been well-characterized and known to be T-cell responses.[115]
highly biocompatible. The size of microparticles can be Liposomes are vesicles composed of naturally occur-
easily controlled, and microparticles of less than ring or synthetic phopholipids. The bilayer structure
100 mm in diameter can be easily administered by injec- can be single- or multicompartment. The size can also
tion through standard-sized needles (22 gauge or smal- vary from smaller than 1 mm to larger than 10 mm.
ler). Because of the slow degradation of the polymer, When negatively charged lipid molecules, which form
antigens are slowly released from the microparticles liposomes, interact with divalent cations, a solid, multi-
for long term in much the same way as do alum adju- layered, crystallaine structure called cochleate is
vants, and this results in enhanced immune responses. formed. Because liposomes and cochleates can protect
Other polymers, such as chitosan, have been used for antigens from the GI tract and deliver them to the
preparation of vaccine formulations. Because one of Peyer’s patches, they have been exploited as an effec-
the important roles that microparticles play in immuni- tive delivery system for oral vaccination.
zation is the slow release of antigens, a number of Liposomes, like other vaccine-delivery systems, can
approaches have been tried to achieve antigen release exert immunoadjuvant effects. The surface charge of
at desired rates. The surface of microparticles can be liposomes is known to affect the immune responses.
modified to alter the adsorption and desorption Positively charged liposomes containing soluble anti-
kinetics of antigens. Alternatively, the pore size can gens were reported to function as a more potent
be varied to control the release of antigens from inducer of antigen-specific, cytotoxic T-lymphocyte
microparticles. responses and delayed-type hypersensitivity responses
The size of microparticles is known to play a critical than negatively charged and neutral liposomes con-
role in oral immunization. In addition to protecting taining the same concentrations of antigens.[116] Stu-
antigens from acidic and enzymatic degradation in dies showed that the positively charged liposomes
the GI tract, microparticulates are known to enhance delivered proteinaceous antigens efficiently into the
uptake by M-cells in the Peyer’s patches, and the effec- cytoplasm of the macrophages/antigen-presenting cells
tiveness of the uptake depends on the size of micropar- where the antigens are processed to be presented by
ticles. It is generally thought that microparticles class I MHC molecules to induce the cell-mediated
smaller than 10 mm are preferentially absorbed by immune response.[116]

Unit–Validation
M-cells, and the smaller the size, the better the absorp- Liposomes containing highly immunogenic glyco-
tion. One study using microparticles of different sizes proteins of the Sendai virus on their surface, which are
showed that the efficiency of uptake of 100-nm parti- called fusogenic liposomes, showed enhanced antigen-
cles by the intestinal tissue was 15- to 250-fold higher specific humoral immunity in mice. The levels of antio-
than that of larger size microparticles.[113] In addition valbumin antibody were markedly increased in serum
to the small size, microparticles with more hydro- from mice immunized with OVA encapsulated in
phobic surface property are absorbed better than those fusogenic liposomes. It appears that the fusogenic
with more hydrophilic surface property. There are, liposomes function as an immunoadjuvant in inducing
however, no definite studies confirming or supporting antigen-specific antibody production.[117]
these assumptions. Once microparticles are placed in Virosomes are liposomes containing viral fusion pro-
the GI tract, adsorption of numerous proteins and teins that allow efficient entering into cells fusion with
polysaccharides present in the GI tract would alter endosome membranes. Viral fusion proteins become
the surface chemistry drastically, and it is difficult to activated in the low pH environment in the endosome
correlate a particular surface chemistry of the native to release its contents into the cytosol.[118] Hepatitis A
microparticles with the absorption ability. and influenza vaccines constructed on virosomes elicited
Virus-like particles (VLPs) consist of one or more fewer local adverse reactions than did their classic
viral-coat proteins. They are very immunogenic mole- counterparts and displayed enhanced immunogenicity.
cules that allow for covalent coupling of the epitopes Virosome-formulated influenza vaccine has also been
of interest.[114] Recently, parvovirus-like particles have shown to be safe and immunogenic when administered
been engineered to express foreign polypeptides in cer- by the intranasal route.[119] Other studies have suggested
tain positions, resulting in the production of large that immunopotentiating reconstituted influenza viro-
quantities of highly immunogenic peptides, and to somes can be a suitable delivery system for synthetic
 3922 Vaccines and Other Immunological Products

peptide vaccines. The virosomes have a great potential costimulatory molecules, and the use of CpG DNA,
for the design of combined vaccines targeted against and cytokines.
multiple antigens and multiple pathogens.[120]
Micelles are aggregates of detergent molecules in
Costimulation
aqueous solution. Detergents are water-soluble, surface-
active agents composed of a hydrophilic head group
Activation of T-cells requires two signals. The first sig-
and a hydrophobic or lipophilic tail group. They can
nal is provided by recognition of MHC/peptide com-
also align at aqueous/non-aquous interfaces, reducing
plex by the T-cell receptor. This does not result in
surface tension, increasing miscibility, and stabilizing
proliferation and differentiation of the T-cell unless
emulsions. Polymeric micelles made of block copolymers,
the T-cell receives a second, costimulatory signal.
such as poly(ethylene oxide)-poly(propylene oxide)-
Several costimulatory signals have been identified, but
poly(ethylene oxide), have been used as a delivery system
the major costimulatory signal appears to result from
for hydrophobic drugs. They can also encapsulate
the binding of CD28 on T-cells to B7 molecules on
antigens for vaccination.
antigen-presenting cells. There are at least two B7
Niosomes are non-ionic surfactant vesicles. They
molecules, B7-1 (CD80) and B7-2 (CD86). Activation
have been used to develop a vaccine-delivery system
of antigen-presenting cells results in increased expression
by peroral and oral routes. Ovalbumin was encapsu-
of B7-2, followed by B7-1. A second T-cell ligand of
lated in various lyophilized niosome preparations con-
the B7 molecules is cytotoxic T lymphocyte antigen-4
sisting of sucrose esters, cholesterol, and dicetyl
(CTLA-4 or CD152) that, other than its name implies,
phosphate. Encapsulation of ovalbumin into niosomes
is rapidly expressed on both CD4þ and CD8þ T-cells
consisting of 70% stearate sucrose ester and 30% pal-
after binding of the T-cell receptor to the MHC/
mitate sucrose ester (40% mono-, 60% di/triester)
peptide complex on antigen-presenting cells. However,
resulted in a significant increase in antibody titers in
in contrast to the positive signal provided by CD28,
serum, saliva, and intestinal washings.[121]
CTLA-4 downregulates T-cell responses.[123] CTLA-4
Cross-linked protein crystals have been used as
has a higher affinity for the B7-molecules than does
antigens. The immunogenicity of cross-linked protein
CD28 and may prevent the activation of T-cells when
crystals of human serum albumin was 6- to 30-fold
B7 expression by dendritic cells is low and terminate
higher in antibody titer than that of the soluble protein
the immune response when its expression is strongly
over an almost 6-month study.[122] It is likely that the
increased. A soluble chimeric protein, CTLA4Ig,
cross-linked protein crystals release antigen in a slow-
blocks the binding of both CD28 and CTLA-4 to the
release manner, and in this sense, the cross-linked
B7 molecules and, thus, may prevent T-cell activation.
protein crystals function as a depot. The cross-linked
Administration of this protein to patients with pso-
protein crystals present high stability, purity, biode-
riasis vulgaris, an immune-mediated skin disease, in a
gradability, and ease of manufacturing, all of which
phase I clinical trial resulted in significant improve-
Unit–Validation

are highly attractive features for vaccine formu-

ment in approximately 50% of the patients.[124] Selec-
lation.[122] Because the cross-linked protein crystals
tive inhibition of CTLA-4 with specific antibodies
are microparticulates, they can also be used for
may boost the immune system. The combination of
vaccination through various routes.
surgery and anti-CTLA-4 antibody therapy was highly
effective in the prevention of metastatic recurrence in a
mouse prostatic carcinoma model.[125]
IMMUNOMODULATION
Other CD28 and B7 homologs continue to be iden-
tified and appear to play a role in costimulation.[126]
Immunomodulation refers to treatments that alter
These molecules may provide additional targets for
immune responsiveness in a non-antigen-specific man-
immunomodulation and suggest that it may be possible
ner. Enhancement of the immune response is desired in
to fine-tune the immune response through pharmaco-
the treatment of chronic infectious diseases and neo-
logic intervention.
plastic diseases, whereas suppression is needed in cases
of inappropriate or exaggerated immune response,
including allergies and autoimmune diseases. There CpG DNA
are numerous treatments that affect the activity of
the immune system. The effect of currently available Bacterial DNA has a higher content of the CpG dinu-
immunosuppressive drugs is very broad, giving these cleotide than does vertebrate DNA, and, in contrast
drugs undesirable side effects. The aim of the research to vertebrate DNA, the CpG is not preferentially
in this area is to design treatments that selectively methylated. The unmethylated CpG DNA sequences
enhance or suppress immune responses. Some of the provides a strong stimulus for the immune system.[127]
newer treatment options are those that target CpG DNA stimulates the secretion of IL-12 by
Vaccines and Other Immunological Products 3923

macrophages and dendritic cells and thus provides a reduces the rate of exacerbation of relapsing-remitting
potent stimulus for type 1 immune responses. It also multiple sclerosis.[128,129] The mode of action of
directly stimulates B cells to proliferate and differen- interferon-b has not been determined. Interferon-b
tiate into immunoglobulin secreting cells. A cellular reduces the production of tumor necrosis factor-a
receptor for CpG DNA has not been identified. The and increases the secretion of IL-10 in vitro. TNF-a
DNA appears to enter the cell via endocytosis, and is a proinflammatory cytokine that may contribute to
some of the DNA escapes the endosomes into the demyelination in multiple sclerosis. IL-10 suppresses
cytoplasm of the cell where it activates various macrophage function and the production of TNF-a.
signaling pathways. In addition, interferon-b may reduce the entry of leu-
Applications for oligonucleotides containing kocytes into the central nervous system, a critical
unmethylated CpG sequences (CpG–ODN) are being component in the inflammation that causes the lesions
explored in various areas of immunotherapy. Adminis- in multiple sclerosis.
tration of CpG–ODN to mice protected against sub-
sequent challenge with the intracellular bacteria Tumor necrosis factor-a inhibitors
Listeria monocytogenes and the intracellular protozoa
Leishmania major. In addition, the CpG–ODN cured Tumor necrosis factor-a (TNF-a) is a cytokine with
established L. major infections. The strong type 1 multiple biological effects. It is produced as a trans-
immunostimulatory property of CpG–ODN makes membrane precursor molecule by various cells in the
this compound a good candidate for vaccine adjuvants. body. It is cleaved by the TNF-a-converting enzyme
Indeed, coadministration of CpG–ODN with antigen and forms trimeric aggregates that bind to either the
markedly boosts the humoral and cell-mediated TNF-receptor (TNFR) I or the TNFR II that are
immune responses. Allergic diseases such as asthma expressed on many different types of cells. The extra-
and atopic dermatitis are caused by type 2 immune cellular domains of the TNFR can be cleaved by
responses directed against otherwise innocuous anti- enzymes and can inhibit TNF-a activity by preventing
gens. Treatment with CpG–ODN cleared established binding of TNF-a to cell-bound receptors. Recent
disease in a mouse model of airway hyper-reactivity, studies have demonstrated that inhibition of TNF-a
suggesting a CpG-induced reversal to type 1 immune activity resulted in significant improvement of the clini-
responses. CpG DNA may also have a place in immu- cal condition of many patients with rheumatoid
notherapy of cancer because of its ability to activate arthritis and inflammatory bowel disease.[130,131] These
NK cells through the induction of IL-12. Administra- studies clearly demonstrate an important role of TNF-
tion of CpG–ODN in combination with monoclonal a in rheumatoid arthritis and inflammatory bowel dis-
antibodies directed against tumor antigens greatly ease, although the precise mechanisms remain to be
enhanced the survival of mice that had been inoculated determined. The inhibition of TNF-a activity is
with tumor cells. achieved by treatment with anti-TNF-a monoclonal

Unit–Validation
antibodies or with soluble TNFR-fusion protein. To
reduce the induction of antibodies against the mouse
Cytokines monoclonal antibodies, the monoclonal antibodies
are chimeric (i.e., the constant portion is derived from
Cytokines play a critical role in the regulation of the human immunoglobulins and the TNF-a-specific vari-
immune and inflammatory response, and they are poten- able portion is derived from mice) or humanized (all of
tial targets for therapy. Important limitations, however, the immunoglobulin is human except for the comple-
are the pleiotropy and redundancy in the cytokine system mentarity determining regions that fold into the
and the short half-life and short action range of most TNF-a-binding region). The TNFR-fusion protein is
cytokines. In spite of these limitations, considerable effort constructed from the extracellular domain of TNFRII
is spent on developing reagents that either block or and the Fc portion of human immunoglobulins. This
enhance the activity of a specific cytokine. construct has a much longer half-life than does the
Two remarkable successes of cytokine therapy are naturally occurring soluble TNFR.
the treatment of multiple sclerosis with interferon-b
and the treatment of rheumatoid arthritis and inflam-
matory bowel disease with tumor necrosis factor-a CHALLENGES IN FUTURE VACCINE
inhibitors. FORMULATIONS

Interferon-b Recent advancements in microbial pathogenesis,

immunology, genetic engineering, plant genetics, and
Clinical trials have demonstrated that subcutaneous expression vector technology have formed the foun-
injections of recombinant or natural interferon-b dation for a new generation of vaccines and other
 3924 Vaccines and Other Immunological Products

pharmaceutical products. New developments in the 5. Murphy, B.R.; Prince, G.A.; Walsh, E.E.; Kim, H.W.;
delivery system have provided us with novel ways to Parrott, R.H.; Hemming, V.G.; Rodriguez, W.J.;
Chanock, R.M. Dissociation between serum neutralizing
enhance the immunogenicity of subunit antigens or and glycoprotein antibody responses of infants and
nucleic acids by their controlled release and reduced children who received inactivated respiratory syncytial
degradation. virus vaccine. J. Clin. Microbiol. 1986, 24, 197–202.
6. Connors, M.; Kulkarni, A.B.; Firestone, C.Y.; Holmes,
For more convenient and more effective immuni- K.L.; Morse, H.C., III; Sotnikov, A.V.; Murphy, B.R.
zation, current vaccine-delivery technologies need to Pulmonary histopathology induced by respiratory syncy-
be improved. Currently, vaccination of many inacti- tial virus (RSV) challenge of formalin-inactivated RSV-
immunized BALB/c mice is abrogated by depletion of
vated or subunit antigens requires booster doses CD4þ T cells. J. Virol. 1992, 66, 7444–7451.
because of the lack of inherent immunogenicity found 7. Top, F.H., Jr.; Buescher, E.L.; Bancroft, W.H.; Russell,
in the natural organism. Thus, reducing the number of P.K. Immunization with live type 7 and type 4 adenovirus
vaccines. II Antibody response and protective effect
doses is one of the primary goals in vaccination. The- against acute respiratory disease due to adenovirus type
oretically, various controlled-release technologies can 7. J. Infect. Dis. 1971, 124, 155–160.
be used to release antigens over time in a sustained 8. Sabin, A.B.; Boulger, L. History of sabin attenuated polio-
virus oral live vaccine strains. J. Biol. Standardization
or pulsatile manner and to direct antigens to specific 1973, 1, 115–118.
antigen-presenting cells for increased vaccine efficacy. 9. Parkman, P.D.; Meyer, H.M., Jr.; Kirschstein, R.L.;
In addition to controlled-release technology, the sin- Hopps, H.E. Attenuated rubella virus. I. Development
and laboratory characterization. New Engl. J. Med.
gle-shot vaccination requires development of better 1966, 275, 569–574.
adjuvants. The mechanism of action of such adjuvants 10. Plotkin, S.A.; Farquhar, J.; Katz, M.; Ingalls, T.H. A new
should be known so that reproducible results can be attenuated rubella virus grown in human fibroblasts:
evidence for reduced nasopharyngeal excretion. Am.
obtained in a mass vaccination program. The require- J. Epidemiol. 1967, 86, 468–477.
ments and problems of immunizing immunocompro- 11. Bunyak, E.B.; Hilleman, M.R.; Weiber, R.E.; Stokes, J.,
mised, immature, older, or pregnant hosts need to be Jr. Live attenuated rubella virus vaccines prepared in duck
embryo cell culture. I. Development and clinical testing.
addressed effectively. Further improvement in our J. Am. Med. Assoc. 1968, 204, 195–200.
understanding of how to modulate Th1 and Th2 12. Morrow, C.D.; Novak, M.J.; Ansardi, D.C.; Porter, D.C.;
responses effectively would certainly help us design Moldoveanu, Z. Recombinant viruses as vectors for muco-
sal immunity, defence of mucosal surfaces: pathogenesis,
better vaccines. Another means of improvement is to immunity and vaccines. In Current Topics in Microbiology
combine a number of vaccines into multivalent vac- and Immunology; Kraehenbuhl, J.P., Neutra, M.R., Eds.;
cines. This will improve the immunization compliance Springer-Verlag: Berlin, 1999; 236, 255–273.
13. Muster, T.; Ferko, B.; Klima, A.; Purtscher, M.; Trkola,
in people living in developed or developing countries. A.; Schulz, P.; Grassauer, A.; Engelhardt, O.G.; Garcia-
Because the majority of pathogens enter their hosts Sastre, A.; Palese, P., et al. Mucosal model of immuni-
via mucosal routes, the new-generation vaccines zation against human immunodeficiency virus type 1 with
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 Validation of Pharmaceutical Processes
Robert A. Nash
Consultant, Mahwah, New Jersey, U.S.A.

INTRODUCTION PROCESS VALIDATION OPTIONS

Process validation is a requirement of current Good The guidelines on general principles of process vali-
Manufacturing Practices (GMPs) for finished pharma- dation[1] mention three options: prospective process
ceuticals (21 CFR 211) and of the GMP regulations for validation (also called premarket validation), retro-
medical devices (21 CFR 820) and therefore applies to spective process validation, and revalidation. Actually
the manufacture of both drug products and medical there are four, if concurrent process validation is
devices. included.
According to the FDA Guidelines on General Prin- Prospective validation is carried out prior to the dis-
ciples of Process Validation,[1] process validation is tribution of a new product or an existing product made
defined, ‘‘ as establishing documented evidence, which under a revised manufacturing process where such
provides a high degree of assurance, that a specific pro- revisions may affect product specifications or quality
cess will consistently produce a product meeting its characteristics. The prospective approach features
predetermined specifications and quality characteris- critical step analysis in which the unit operations are
tics.’’ The process for making a drug product consists challenged during the process qualification stage to
of a series (flow diagram in logically defined steps) of determine those critical process variables that may affect
unit operations (modules) that result in the manufac- overall process performance, using either worst-case
ture of the finished pharmaceutical. analysis or a fractional–factorial design. During formal,
There is much confusion regarding the definition of three-batch, prospective validation, critical process
process validation and what constitutes process vali- variables should be set within their operating ranges
dation documentation. The term validation is used here and should not exceed their upper and lower control lim-
generically to cover the entire spectrum of current GMP its during process operation. Output responses should be
concerns, essential most of which are; facility, equip- well within finished-product specifications.
ment, component, method, and process qualification. Retrospective validation is recognized in both
Based upon the FDA process validation guidelines,[1] current GMPs [21 CFR 211.110(b)] and the FDA pro-
cess validation guidelines.[1] It involves accumulated
Unit–Validation

the specific term should be reserved for the final stage(s)

of the product and process development sequence. in-process production and final product testing and
The essential or key steps or stages of a successfully control (numerical) data to establish that the product
completed development program are shown in Table 1. and its manufacturing process are in a state of control.
The end of the development sequence, which should Valid in-process results should be consistent with the
be assigned to formal (three-batch) process validation, final specifications of the drug product and shall be
derives from the fact that the specific exercise of pro- derived from previous acceptable process average and
cess validation should never be designed to fail. Failure process variability estimates where possible and deter-
in carrying out the formal process validation assign- mined by the application of suitable statistical procedures
ment is often the result of incomplete or faulty under- (quality control charting) where appropriate.
standing of the process capability, in other words, what The retrospective validation option is chosen for
the process can and cannot accomplish under a given established products whose manufacturing processes
set of operational requirements. are considered to be stable and when, on the basis of
In a well-designed validation program, most of the economic considerations and resource limitations,
effort should be spent on facilities, equipment, compo- prospective qualification and validation experimen-
nents, methods, and process qualification. In such a tation cannot be justified. Prior to undertaking either
program, the formalized, final three-batch validation prospective or retrospective validation, the facilities,
sequence provides only the necessary process vali- equipment, and subsystems used in connection with
dation documentation required by the FDA to show the manufacturing process must be qualified in con-
product reproducibility and a manufacturing process formance with cGMP requirements.
in a state of control. Such a strategy is consistent with Concurrent validation studies are carried out under
the FDA preapproval inspection program directive.[2] a protocol during the course of normal production.
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-100001927
3928 Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved.
Validation of Pharmaceutical Processes 3929

Table 1 Development sequence with respect to process 4. Change in a facility and/or plant (usually
validation location, site, or support systems);
Developmental stages Batch size 5. Significant increase or decrease in batch size
Product design 1X
that affects the operation of modular equip-
Product characterization ment; and
Product selection 6. Sequential batches that fail to meet product and
Process design process in-process specifications.
Product optimization 10X
Process characterization In some situations process performance requalifica-
Process optimization tion studies may be required prior to undertaking
Process qualification specific revalidation assignments. With the exception
Process qualification 100X of sterile products manufacture, periodic revalidation
Process validation is not required at the present time. The performance
Process certification
and state of control of the product and its manufactur-
Process revalidation 100X to 1000X
ing process can be adequately covered during the
annual product and process review. The FDA has
issued an interim guidance document that addresses
what constitutes major and minor formulation and
The first three production-scale batches must be mon- manufacturing changes for immediate-release solid
itored as comprehensively as possible. The evaluation dosage forms.[4] Such documentation and others to fol-
of the results is used in establishing the acceptance low should simplify manufacturing decisions about the
criteria and specifications of subsequent in-process need to revalidate.
control and final product testing. Some form of
concurrent validation, using statistical process con-
trol techniques (quality control charting) may be
employed throughout the product manufacturing life VALIDATION PRIORITIES
cycle.
Revalidation is required to ensure that changes in the There is a basic concept with respect to which pharma-
process and/or in the process environment, whether ceutical processes should be given a higher priority
introduced intentionally or unintentionally, do not over others. All pharmaceutical manufacturing pro-
adversely affect product specifications and quality cesses require process validation documentation, but
characteristics.[2] There should be a quality assurance there is an accepted logical approach to priority selec-
program (change control) in place which requires tion, in the following order:
revalidation whenever there are significant changes in Sterile products and their processes

Unit–Validation
formulation, equipment, process, and packaging that
may impact on product and manufacturing process
large volume parenterals (LVPs) infusions greater
performance.[3] Furthermore, when a change is made than 100 ml
in a raw material supplier, the drug manufacturer should
small volume parenterals (SVPs) single and multiple
be made aware of subtle, potentially adverse differences dose injections
in raw material characteristics that may adversely affect
ophthalmics and sterile devices
product and manufacturing process performance.
It is recommended that every requested change be Non-sterile products and their processes
reviewed by the validation or CMC committee. Such
a committee should judge if a change is significant
low dose high potency tablets and capsules
for revalidation and decide on a course of action to
drugs with inherent stability problems
be taken. The following conditions require revalidation
transdermal delivery (TDD) and inhalation pro-
study and documentation: ducts

the rest of the oral solid dosage forms
1. Change in a critical component (usually refers
oral liquids and topical products
to active pharmaceutical ingredient, key excipi-
ents, or primary packaging); The best approach to assessing problems with
2. Change or replacement in a critical piece of respect to a terminal sterilization method (i.e., moist
modular (capital) equipment; heat, dry heat, radiation, and chemical methods) is to
3. Significant change in processing conditions that first establish the qualification, validation and stability
may affect subsequent unit operations and pro- of the pharmaceutical process prior to conducting a
duct quality; given sterilization procedure.
 3930 Validation of Pharmaceutical Processes

Table 2 Composition of the process validation committee

Representative of Function
Engineering Qualifies for plant, facilities, equipment, and support systems
Development Qualifies for products and their specific manufacturing processes
Manufacturing Operates plant, facilities, equipment, support systems and the various manufacturing processes
Quality assurance Audits plant, facilities, equipment, support systems, the various manufacturing
processes, and their products

THE VALIDATION COMMITTEE validation program. A Process Validation progress

chart is shown in Fig. 1.
In most companies, the validation or Chemistry,
Manufacturing and Control (CMC) committee is
charged with the responsibility of establishing and Installation Qualification (IQ)
operating the complete validation program for the
specific manufacturing site. In some companies the This includes procedures and documentation to show
program is led by a validation manager whereas in that all important aspects of the installation of the
others, quality assurance personnel have taken on facility, support system, or piece of modular equip-
expanded responsibilities in this regard. ment, having been properly calibrated, meet its design
Specific process validation assignments are carried specifications and that the vendor’s recommendations
out by those with the necessary training and experi- had been suitably considered.
ence. The specifics of how the committee is organized
to conduct process validation assignments is beyond
the scope of this article. The responsibilities that must Operational Qualification (OQ)
be carried out and the traditional organizational struc-
tures best equipped to handle each of these assign- Following IQ, procedures and documentation show
ments are outlined in Table 2. Other members may that the facility, support system, or piece of modular
include Quality Control, Regularity affairs, etc. equipment perform as intended throughout all antici-
pated operating ranges under a suitable load.

VALIDATION MASTER PLAN

Unit–Validation

The creation of a master plan permits the development

of a logical overview of the validation effort. It lays out
in a logical sequence the activities or key elements or
both to be performed in accordance with the approxi-
mate time schedule in a Gantt or PERT chart format.
The master plan establishes the critical path through
the chart against which progress can be monitored.
The validation program starts with the design and
development of raw materials and components, fol-
lowed by the IQ/OQ of facilities, equipment, and sys-
tems through performance and process qualification
stages, and terminates in the protocol-driven, three-
batch, formal process validation program. Most of
these activities move forward in series. However, by
combining activities and elements and moving in paral-
lel, where possible, on independent tracks with respect
to active pharmaceutical ingredients (APIs) analytical
methods development, facilities, equipment, support
systems, and the drug product design and manufactur-
ing process development, a great deal of time can be
saved before the individual elements or grouping of
activities are combined prior to the formal process Fig. 1 Process validation progress chart.
Validation of Pharmaceutical Processes 3931

Performance Qualification (PQ) accuracy of reported and factual information in New

Drug Application (NDA) and Abbreviated New Drug
Following IQ and OQ, actual demonstrations during the Application (ANDA) submissions with respect to the
course of the validation program show that the facility, suitability of cGMP product development, analytical
support system, or piece of modular equipment perform laboratory, and manufacturing facilities.
according to a predefined protocol and achieve process A preapproval inspection checklist should include
reproducibility and product acceptability. the following documentation which may be required
prior to the formal inspection:
VALIDATION PROTOCOL AND REPORT
1. API development and validation report(s)
including impurity profile and polymorphic
The following validation protocol and format for the
forms;
completed validation report have been suggested in
2. Pharmaceutical (dosage form) development
the WHO Guidelines on Validation of Manufacturing
report;
Processes (TRS 823).[5]
3. Stability and clinical batch records and history,
including phase-III program;
1. Purpose (for the whole validation) and prereq-
4. Data for API and key excipients used in the
uisites
manufacture of clinical and biobatches;
2. Presentation of the whole process and subpro-
5. Bioequivalency report;
cesses including flow diagram and critical step
6. Technical transfer report (development to
analysis
manufacturing/QA/QC);
3. Validation protocol approvals
7. Copy of the CMC section of the NDA including
4. Installation and operational qualifications,
information on suppliers and vendors;
including blueprints or drawings
8. Copy of proposed production monograph and
5. Qualification report(s)
master batch record;
9. Equipment validation report establishing IQ

Subprocess 1
and OQ;

Purpose
10. Cleaning validation report;

Methods and procedures
11. Analytical methods validation and computer

Sampling and testing procedures, release cri-
systems validation reports;
teria
12. Stability report establishing expiry dating; and

Reporting function
13. Process validation protocol for formal three-

Calibration of test equipment
batch validation of production-size batches.

Test data

Summary of results

Unit–Validation
During preapproval inspection, the FDA accepts a

Approval and requalification procedure
process-validation protocol based on the company’s

Subprocess 2 (repeat)
commitment to complete successfully three production-
size validation batches prior to product launch. In
6. Product qualification, test data from prevalida-
some situations a prevalidation (process demonstration
tion batches
qualification) production-size batch is completed before
7. Product validation, test data from three formal
the entire formal three-batch program is carried out.
validation batches
8. Evaluation and recommendations (include reva-
lidation and requalification requirements)
PILOT SCALE-UP AND TECHNOLOGY
9. Certification (approval)
TRANSFER
10. Summary report with conclusions
The pilot-production program may be carried out as a
The validation protocol and report may also include
shared responsibility between the development labora-
copies of the product stability report or its summary as
tories and their appropriate manufacturing counter-
well as validation documentation on cleaning and ana-
part or as a process demonstration by a separate,
lytical methods.
designated pilot-plant or process development depart-
ment. Supporting technology transfer documentation
PREAPPROVAL INSPECTION applies to both the specific process and system being
qualified and validated and the related testing
The FDA Preapproval Inspection Program[2] is designed standards and testing methods. The formal technology
to provide a basis for determining the adequacy and transfer is normally made from the development
 3932 Validation of Pharmaceutical Processes

Establish chemical and physical compatibility

Minimize lot-to-lot variability in properties

Worldwide availability from comparable suppliers

Properties for possible evaluation
Fig. 2 Technology transfer stages.
– Color, odor, taste, solubility;
laboratories or the process development pilot-plant to – Particle morphology (DSC, TGA, x-ray diffrac-
pharmaceutical production function. tion);
In actuality, a number of technology transfer points – Particle size distribution and surface area;
and documents are generated as prospective validation – Crystal and bulk density, compaction index;
proceeds through the various stages of product devel- – Angle of repose and flowability index;
opment. These stages of technology transfer in terms – Spectrophotometry (UV, FTIR, NMR, OR);
of scale-up are illustrated in Fig. 2. – Water content, LOD, moisture uptake;
Solid pharmaceutical dosage forms (tablets and – Microbial limits and heavy metals;
capsules) are used to illustrate the various stages of – HPLC assay and impurity profile.
product and process development. These principles
and practices also apply in a general way to the devel- Before preformulation studies are undertaken,
opment of liquid and semisolid pharmaceutical dosage two-way technical communication between the manu-
forms (not discussed here). facturers of the API (laboratory and plant) and the
pharmaceutical product development laboratories
must be established (Fig. 1). It should start early and
STAGES OF VALIDATION be maintained through-out the product and process
development program.
Elements of the validation concept should be incorpo- In addition to potency, purity, and stability consid-
rated during each of the various stages of the product erations of the API, the product development depart-
and process development continuum. These stages can ment is especially interested in the chemical and
be summarized as follows. physical form (free acid or base, salts, esters, amides,
polymorphs, solvates, particle size and shape) of the
API. Time spent early in the cycle in establishing these
Stage Preformulation Studies: particular factors often aids and/or simplifies the sub-
APIs plus rey excipients sequent product and process development program.
Not every subject shown above must be tested or
Stage I Product design and development addressed. However, aspect, particle morphology and
Stage II Preparation of clinical and biobatches size, compaction and flowability, water content, spec-
Unit–Validation

Stage III Process scale-up and evaluation trophotometric and chromatographic data should be
Stage IV Formal process validation studied and monitored throughout the product and
process development program.[6,7]
Because key excipients are well established in most
Preformulation Studies: API new product and process development programs, the
same degree of preformulation scrutiny is often not
Preformulation testing of the specific API of interest required. Compatibility studies with the API, however,
and key excipients to be used in the product design should be performed to study possible untoward inter-
stage, alone and in combinations with the API, should actions between the active ingredients and the excipi-
be included as a preliminary first step in the product ents. It should be kept in mind that small or minor
and process development sequence. A simple check list changes in physical and possibly chemical properties
of items worth consideration in preformulation studies upon intimate contact in binary studies with key exci-
with APIs and important or critical excipients is pro- pients should not automatically exclude a favored
vided as follows: excipient without further critical testing.
API

Key excipients Stage I: Product Design and Development

– Fillers and diluents Following successful preformulation studies, the API is

– Binders transferred to the formulations laboratory for prelimi-
– Disintegrants nary product design and development studies. In most
– Glidants and lubricants cases, the drug is mixed with an appropriate diluent or
Validation of Pharmaceutical Processes 3933

filler and glidant combination and filled into two-piece upon accelerated, elevated temperature testing (1 month
opaque hard-shell capsules for preliminary stability at 45 C or 3 month at 40 C and 80% relative humidity)
and subsequent phase I clinical studies versus matching the next step (stage II) is to scale the product and its
placebo capsules.[8] At or about the same time, initial process to (10X) pilot-laboratory size batch(es). This
studies of a prototype tablet formulation should be batch represents the first replicated scale-up of the
started. The key steps in the product design and devel- designated formula. Its size usually ranges between
opment sequence are given below. 10 and 100 kg, 10 and 100 L, or 10,000 to 100,000 units.
Stage I: Product Design: 1 Laboratory Scale Often these pilot-laboratory batches are used in clinical
(1–10 kg) trials and bioequivalency studies. According to the
FDA, the minimum requirement for a biobatch is

Hard-shell capsule (phase I clinical trials) followed 100,000 units.[10]
by prototype tablet dosage form Pilot-laboratory batches are usually prepared in
small pilot equipment within a designated current
– Direct compression versus wet granulation GMP approved facility. The number and size of these
– Maximize chemical and physical stability pilot-laboratory batches may vary, depending on one
– Minimize product and process costs or more of the following factors:
– Product characterization
– Product selection
Equipment availability
– Process design
API availability

Cost of raw materials

Excipients are selected among the following cate-
Inventory requirements for both clinical and non-
gories: clinical studies
Binder, diluents, and disintegrants including
alginates, calcium phosphate, cellulose, dextrates, Process development (process qualification) or pro-
gelatin, povidones, starch and derivatives, sorbitol, cess capability studies are normally started in this impor-
sucrose, and derivatives. Glidants and lubricants tant stage II of the scale-up sequence. The scope of
including colloidal silicon dioxide, hydrogenated stage-II process development consists essentially of pro-
vegetable oil, mineral oil, PEG, silica gel, sodium duct optimization and process characterization studies.
lauryl sulfate, stearates, talc. Product Optimization:

Although the work is conducted in the research or
Establish formula rationale and boundary con-
formulations laboratory using small-scale processing ditions for API and excipients
equipment, it is important to gain early experience with Process Characterization:
colorant systems that have been selected for the fin-

Define unit operations, process variables, and

Unit–Validation
ished tablet product; color aids in blend-uniformity
evaluation. response parameters.
In addition to excipient screening and selection, it is
important to gauge processing parameters that are – Define critical process variables and response
more fully explored during the scale-up phases. These parameters using simple experimental designs.
processing factors include flowability, compaction – Establish provisional control limits for critical
and compressibility of powders and granules, content process variables and their response parameters
uniformity of powder and granule blends and finished based on process replication.
tablets, moisture uptake, in vitro dissolution release
profiles, and subsequent full-scale stability testing.
Maintain product stability.
Products used in human clinical trials must, of
course, conform to good laboratory, good clinical, Unit operations for solid dosage-form development
and good manufacturing practice requirements.[1,9] include:

Granulation

Drying
Stage II: Process Development:
Sizing
Pilot Laboratory (Clinical)
Blending and mixing

Encapsulation andar tablet compression
After the (1X) ‘‘go’’ laboratory batch has been deter-
Coating
mined to be both physically and chemical stable, based
Filling and packaging
 3934 Validation of Pharmaceutical Processes

Unit operations are selected for the development of Fahrner[17] raises the following issues regarding the
a tablet (coated or non-coated) or capsule (hard shell new role for pilot plants in product development.
or soft-gel) process.[11] Unit operations that are con-
sidered to be critical are determined through analysis

Too much time is devoted to preliminary or applied
of the process variables and their respective measured
research and not enough to the proper development
response for each unit operation (Table 3).[12–14]
of the process.
In order to determine critical control parameters and

Often a suitable manufacturing strategy is lacking
their unit operations, constraint analysis techniques[15]
during the early phases of the program, which
followed by fractional factorial designs (Table 4) are
results in poorly planned technology transfer and
used to challenge the tentative control limits (so-called
an inappropriate division of responsibility with
worst-case analysis) established for the process at this
respect to the overall program.
intermediate stage. Time and effort spent to qualify

Most laboratory processes are rarely scalable, since
the process at the 10X stage often simplifies the work
piloting is a scaled-down version of manufacturing
that follows during stages III and IV.
not a scaled-up version of the laboratory batch.
Von Doehren et al.[13] and Chowhan[16] have
described the various stages of solid dosage form pro-
cess development as it relates to technology transfer Fahrner makes the case for a separate pilot facility
and process validation. Their respective approaches (process development function) to bridge the com-
to the topic have been integrated in this article. munication gap between R & D and production.

Table 3 Control parameters for solid-dosage-form development

Unit Operation Process variables (X)a Measured responses (Y)b
Granulation (Power type) Load Power consumption
Speed (main chopper)
Liquid addition rate
Granulation time
Drying Load Moisture content
Inlet temperature Bulk density
Air-flow rate
Drying time
Sizing (Screening) Load Particle size distr.
Screen size Bulk density
Speed
Unit–Validation

Feed rate
Blending (Mixing) Load Blend uniformity
Speed
Mixing time
Encapsulation Fill volume Capsule weight
Tamper setting Moisture content
Speed Dissolution
Glidant (type, amount) Content uniformity
Potency
Tablet compression Press speed Tablet weight
Feed rate Moisture content
Precompression force Hardness/friability
Compression force Thickness
Dissolution/disintegration
Content uniformity
Potency
Coating (Film type) Load Weight gain
Pan speed
Spray rate
Air flow
a
7–23 possible variables.
b
11–16 possible responses.
Validation of Pharmaceutical Processes 3935

Table 4 Fractional factorial design for process equipment. The technology transfer documents should
developmenta include the technical information normally required
Key variablesb for preapproval inspection:

Trials X1 X2 X3 X4 X5 X6 X7 Sums
1        0/7 1. Preformulation information
2    þ    1/6
2. Product development report
3. Product stability report
3   þ   þ  2/5
4. Analytical methods report
4 þ þ   þ   3/4 5. Proposed manufacturing formula, manufactur-
5 þ þ    þ þ 4/3 ing instruction, in-process and final product spe-
6 þ  þ þ þ  þ 5/2 cifications at the 100-batch size
7  þ þ þ þ þ þ 6/1
8 þ þ þ þ þ þ þ 7/0 The objectives of prevalidation trials at stage III
Sums 4/4 4/4 4/4 4/4 4/4 4/4 4/4 28/28 (100 pilot production) is to qualify and optimize
a
Adapted from Hendrix, C. D., What every technologist should the process in full-scale production equipment and
know about experimental design, CHEMTECH (March 1979). their facilities.
b
Key variables are randomly assigned an ‘‘X’’ value.
Rushing through the first (100) pilot-production
batch in order to proceed with formal validation
Stage III: Pilot Production should be discouraged. Small problems that often arise
during (100) scale-up should be addressed immedi-
The technology transfer of the product and process ately and not ignored. Such problems are often best
from the traditional product development function to addressed by returning to the laboratories (10) for
a separate process development (pilot plant) function supplemental process characterization and qualifi-
or production itself is normally carried out at the cation studies.
(100) pilot-production batch stage (100–1000 kg): Many companies, however, proceed directly to
three-batch formal validation without stage III preva-
lidation work and often complete formal trials prior

Full-scale production batch
to preapproval inspection. The downside of this alter-

For possible future commercial or clinical use
native strategy is that finished production batches

Evaluate critical process parameters; product and
often remain in the warehouse beyond their approved
process are scaled to another order of magnitude
expiry dating period.
(100)
When faced with a choice of strategies, there is no

Process optimization
one ideal way of completing the pilot scale-up and

Unit–Validation
validation sequence other than depending on prior
– Mixing and blending times
experience with related products and their processes.
– Drying times
– Milling operations
– Press speed, compression force
– Encapsulation speed, tamping settings Stage IV: Formal Process Validation
– Speed, air flow, spray settings, temperature
In the normal course of events and following a success-

Process qualification (prevalidation batches); deter- fully completed preapproval inspection, formal, three-
mine process capability, challenge in-process con- batch process validation is carried out in accordance with
trol limits the protocol approved during the preapproval inspec-

Maintain product stability tion. The primary objective of the formal process vali-
dation exercise is to establish process reproducibility
The creation of a separate pilot plant or process and consistency. The program is not designed to chal-
development unit has been favored in recent years lenge upper and lower control limits (so-called worst-case
because it is ideally suited to carry out key process analysis) of critical process variables. Such upper and
qualification and/or process validation studies in a lower control limit challenging is normally conducted
timely manner.[18,19] during the stage II (10X size) process characterization,
The objective of the pilot-production batch is to optimization, and qualification program, using suitable
scale the product and its process by another order of and reasonable experimental designs (Table 4).
magnitude (100). For most solid dosage forms it The documentation to be established before, during,
represents a full production scale batch, in standard and after formal process validation is shown below.
 3936 Validation of Pharmaceutical Processes

The protocols and the subsequent formal validation
API changes presently called BACPAC
studies are designed to establish uniformity among
Packaging changes called PACPAC
the three batches with respect to granulation, blending,
Analytical Methods changes called AMPAC
finished tablet, and finished capsule stages.[1,2,10]
Sterile Aqueous Solutions called SUPAC-SAS

100 production batches If the program is successful, other dosage form cate-
gories will be added later.

Complete product development program and report The following changes are covered:

Prepare protocol for prospective process validation

Complete preapproval inspection requirements;
Components and Composition Changes
conduct three-batch formal process validation,
Manufacturing Equipment and Process Changes
establish reproducibility for mixing, blending, and
Batch Size (scale-up) Changes
compression or encapsulation operations
Manufacturing Site Changes

Establish process documentation
The program consists of three levels:
– Preformulation report
– Analytical methods validation report
Level 1 or minor changes that are made without FDA
– IQ/OQ and cleaning validation reports approval and reported in the Annual Report (AR).
– Formula development report
Level 2 or intermediate changes that may be insti-
– Process feasibility report tuted by first filing a Change Being Effected
– Manufacturing bioequivalency report (CBE) Supplement with the FDA and waiting 30
– Product development report days for a reply before instituting the change.
– Process validation protocol
Level 3 or major change in which a prior approval
– Process validation report supplement (PAS) is filed with the FDA and
– Product stability report approval must be obtained from the FDA before
proceeding with the change.

In that respect, the following test data and results This new program is off to a good start and is inter-
are used to show process reproducibility and consist- mittently involved with the need for adequate process
ency among validation batches: particle or granule size validation studies and documentation to support the
distribution, bulk density, moisture content, hardness, changes requested.
thickness, friability, weight uniformity, potency unifor-
mity, disintegration–dissolution profile, and product
stability. Not every one of these categories have to be CHANGE CONTROL
Unit–Validation

addressed nor followed both during in-process and

final product testing. Nevertheless, testing must be Procedures with respect to establishing change con-
sufficient to establish process reproducibility and trol should be in place before, during, and after the
demonstrate, with a high degree of certainly, that the completion of the formal validation program. A change
product and process are in a state of control. control system maintains a sense of functionality as the
Whenever possible, formal validation studies should process evolves and provides the necessary documenta-
continue through packaging and labeling operations tion trail that ensures that the process continues in a vali-
(whole or in-part), so that machinability and stability dated, operational state, even when small non-critical
of the finished product can be established and docu- adjustments and changes have been made. Such minor,
mented in the primary container–closure system. non-critical changes in materials, methods, and machines
Recently the FDA, with the cooperation of the should be reviewed by the validation commitee (develop-
Pharmaceutical Industry has developed a series of ment, engineering, production, and QA/QC) to ensure
guidance procedures to speed approval of post that process integrity and comparability have been
approval changes with or without process scale-up maintained and documented before the specific change
changes. At the present time the SUPAC (scale-up post that has been requested can be approved by the head
approval change) program covers the following pro- of the quality control unit.
duct categories: The change control system, based upon an approved
standard operating procedure(s) (SOPs), takes on

Immediate Release (IR) solid dosage forms added importance as the vehicle or instrument through

Extended Release (ER) solid dosage forms which innovation and process improvements can be

Delayed Release (MR) solid dosage forms made more easily and more flexibly without prior

Semisolids (SS) dosage forms formal review on the part of the NDA and ANDA
Validation of Pharmaceutical Processes 3937

reviewing function of the FDA. If more of the sup- 6. Final determination of conformity to appropri-
plemental procedures with respect to the chemistry ate specifications and justification of the actions
and manufacturing control sections of NDAs and taken, and
ANDAs could be covered through annual review doc- 7. Signature of individual(s) responsible for final
umentation procedures, with appropriate safeguards, decision(s) and the action(s) taken.
process validation will become more innovative.[20–22]
Even though the responsibility for batch acceptance
or rejection lies with the head of the quality control
unit, the help of the validation committee should prove
OUT-OF-SPECIFICATIONS useful in reviewing the process of OOS investigation
and arriving at a recommendation for action taken.
Probably the single most important technical issue
facing the pharmaceutical industry at the present time
is the question: what constitutes process or batch fail- CLEANING VALIDATION
ure in terms of an out-of-specification (OOS) assay
value? The concept of product and/or process failure According to 21 CFR Sect. 211.67, Equipment Cleaning
appears twice in the cGMPs.[3] and Maintenance of cGMP regulations,[3] equipment
According to 21 CFR Sect. 211.165(f), ‘‘Drug pro- and utensils should be cleaned, maintained, and sani-
ducts failing to meet established standards or specifica- tized at appropriate intervals to prevent malfunction
tions and other relevant quality control criteria shall be or contamination that would alter the safety, identity,
rejected.’’ In CFR Sect. 211.192 it is stated: strength, quality, or purity of the drug product. Written
procedures shall be established and followed for clean-
Any unexplained discrepancy (including a percentage
ing and maintenance of equipment. These procedures
of theoretical yield exceeding the maximum or mini-
mum percentages established in master production shall include, but are not limited to the assignment of
and control records) or the failure of a batch or any responsibility for cleaning and maintaining equipment;
of its components to meet any of its specifications shall maintenance, cleaning, and sanitizing schedules where
be thoroughly investigated [regardless] whether the appropriate; description in sufficient detail of methods,
batch has already been distributed. The investigation equipment, and materials used in cleaning and mainte-
shall extend to other batches of the same drug product nance operations, and the methods of disassembling
and other drug products that may have been associated and reassembling equipment as necessary to assure
with the specific failure or discrepancy. A written rec- proper cleaning and maintenance; removal or oblitera-
ord of the investigation shall be made and shall include tion of previous batch identification, protection of clean
the conclusions and follow-up.
equipment from contamination prior to use; and inspec-
tion of equipment for cleanliness immediately before

Unit–Validation
The key to establishing product and/or process fail- use. Records shall be kept of maintenance, cleaning,
ure is to verify the accuracy, relevance, and reproduci- sanitizing, and inspection.
bility of deviant assay value(s), test result(s), and The objective of cleaning validation of equipment and
recorded number(s) that are reported.[23] All companies utensils is to reduce the residues of one product below
should have SOPs in place that cover first the verifi- established limits so that the residue of the previous
cation of deviant numbers in the quality control labora- product does not affect the quality and safety of the sub-
tories and, following that investigation and a report sequent product manufactured in the same equipment.
showing the test result to be deviant, a second set of According to 21 CFR Sect. 211.63, Equipment
SOPs covering follow-up actions taken as described in Design, Size, and Location, of cGMP regulations,[3]
the following steps: equipment used in the manufacture, processing, pack-
ing, or holding of a drug product shall be of appropri-
1. A written procedure for full investigation when ate design, adequate size, and suitably located to
there is not a verified laboratory error. facilitate operations for its intended use and for its
2. Scientific criteria for retesting and resampling cleaning and maintenance. Some of the equipment
during the formal investigation. design considerations include type of surface to be
3. Description and results of the formal investi- cleaned (stainless steel, glass, plastic), use of dispos-
gation into possible causes of the OOS result(s). ables or dedicated equipment and utensils (bags, filters,
4. Results of all testing involved during the inves- etc.), of stationary equipment (tanks, mixers, centri-
tigation. fuges, presses, etc.), of special features (clean-in-place
5. A scientific basis and justification for discarding systems, steam-in-place systems), and identifying the
any OOS test result and accepting the batch in difficult-to-clean locations on the equipment (so-called
question. hot spots or critical sites).
 3938 Validation of Pharmaceutical Processes

The specific cleaning procedure should define the – Sodium starch glycolate
amounts and the specific type of cleaning agents – Starch, pregelatinized
and/or solvents used. The cleaning procedure should
give full details as to what is to be cleaned and how
Tablet and Capsule Lubricants:
it is to be cleaned. The cleaning method should focus
on worst-case conditions, such as highest-strength, – Magnesium stearate
least-soluble, most difficult to clean formulations. – Mineral oil, light
Cleaning procedures should identify the time between – Polyethylene glycol
processing and cleaning, cleaning sequence, equipment – Sodium stearyl fumarate
dismantling procedure, need for visual inspection, and – Stearic acid, purified
provisions for documentation. – Talc
The choice of a particular analytical method – Vegetable oil, hydrogenated
(HPLC, TLC, spectrophotometric, total organic car-
bon (TOC), pH, conductivity, gravimetric, etc.) and
Other Excipients:
sampling technique chosen (direct surface by swabs
and gauze, or by rinsing) depends on the residue limit – Carboxymethylcellulose sodium
to be established, based upon the sampling site, type of – Cellulose acetate phthalate
residue sought, and equipment configuration (critical – Ethylcellulose
sites vs. large surface area) considerations. The analyti- – Hydroxypropyl cellulose
cal and sampling methods should be challenged in – Hydroxypropyl methylcellulose
terms of specificity, sensitivity, and recovery. – Hydroxypropyl methylcellulose pathalate
The established residue limits must be practical, – Methacrylic acid copolymer
achievable, verifiable, and assure safety. The potency – Polysorbates
of selected drug and presence of degradation products, – Polyvinyl acetate phthalate
cleaning agents, perticulates and microorganisms – Povidone
should be taken into consideration. – Sodium lauryl sulfate
The following residue limits have been suggested: not
more than (NMT) 10 ppm or NMT 0.001% of the dose
of any product appears in the maximum daily dose of
another product and no residue visible on the equip-
ment after cleaning procedures have been performed.
Unit–Validation

PHARMACEUTICAL EXCIPIENTS

Pharmaceutical Excipients are components of finished

drug products and site active components are recognized
in the USP/NF. A list of key excipients for solid dosage
forms is given:

Diluents, Fillers, and Binders:

– Calcium phosphate, dibasic

– Dextrates
– Dextrin
– Lactose (anhydrous, fast flow)
– Mannitol
– Starch (corn)
– Sugar, compressible

Tablet Disintegrants:

– Cellulose, microcrystalline
– Croscarmellose sodium Fig. 3 Manufacture of active pharmaceutical ingredients
– Crospovidone (APIs).
Validation of Pharmaceutical Processes 3939

ACTIVE PHARMACEUTICAL a description of the chemistry involved are helpful in

INGREDIENTS (API) defining the process. The process description should
include appropriate parameters, such as charging quan-
A chemical is considered to be an API if it is intended tities or volumes of reactants or solvents, reaction times,
for medicinal purposes. Regulatory agencies however, temperatures, pressures, etc. Critical processing steps
place greater emphasis and priority on the manufac- and critical operating parameters should be maintained
ture and validation of APIs than UPS/NF exciepients. to ensure batch-to-batch consistency, product yield,
According to Sect. 501 (a)[2](b) of the Food, Drug, and quality.
and Cosmetic (FD&C) Act, all drugs must be manu- Where in the chain of unit operations (chemical pro-
factured, processed, packed, and held in accordance cess) does API validation start? As long as key inter-
with cGMPs. No distinction is made between APIs mediates are made in the plant, they and their
and finished drug products. reaction and processing steps should be subjected to
Elements common to both APIs and finished drug an appropriate cGMP and process qualification-
products include facilities and equipment qualification validation program. A key intermediate is defined as an
(IQ/OQ, and PQ), cleaning validation, validation of intermediate in which an essential molecular character-
water supplies, microbial limits for non-sterile material, istic, usually related to stereochemical configuration, is
manufacture of sterile and pyrogen-free material, introduced into the final API structure (moeity).
in-process blending and mixing, analytical methods
validation, laboratory controls and in-process testing,
change control procedures and revalidation, reproces- Physical Characteristics
sing, packaging and labeling, and stability testing.
Besides purity (chemical potency), the physical charac-
teristics and properties of the API are extremely impor-
Process tant to the end user (drug product manufacturer).
Characteristics such as crystal morphology, particle
There are four primary processes used in the manufac- size and shape, bulk density, melting point, optical
ture of APIs. They are chemical synthesis, fermentation, rotation, etc., have a profound effect upon the final
extraction, and purification. A flow diagram (Fig. 3) and drug product and its performance and stability.

Table 5 Meeting quality standards

ISO 9000 series cGMPs
Management responsibility Organization and personnel
Quality system Organization and personnel, laboratory, controls, records, reports

Unit–Validation
Contract review Holding and distribution
Design control Production and process controls
Document control Records and reports
Purchase products Control of components, drug product containers, closures
Supplied product Holding and distribution
Product identification, traceability Packaging and labeling control
Process control Production and process control
Procedures for inspection, testing Laboratory controls
Inspection, measuring, testing equipment Laboratory controls production, and process controls
Inspection, testing status Laboratory controls
Control of nonconforming products Returned and salvaged drug products
Corrective action Records and reports
Handling, storage, packaging, delivery Packaging and labeling control holding, distribution
Quality records Records and reports
Internal quality audits Laboratory controls
Training needs Organization and personnel
Servicing procedures Equipment
Statistical techniques Building, facilities production and process controls
 3940 Validation of Pharmaceutical Processes

In addition to the reaction or extraction step, crystalli- REFERENCES

zation, milling, and blending unit operations must be
subject to qualification and validation. 1. Berry, I.R.; Harpaz, D. Validation of Active Pharmaceutical
Ingredients; Interpharm Press, Inc.: Buffalo Grove, IL, 2001.
2. Carleton, F.J., Agalloco, J.P., Eds.; Validation of Pharma-
ceutical Processes, Sterile Products; Marcel Dekker, Inc.:
New York, 1998.
Impurity Profile 3. Chow, S.-C., Liu, J.-P., Eds.; Validation, Process Controls
and Stability, Statistical Design and Analysis in Pharma-
ceutical Science; Marcel Dekker, Inc.: New York, 1995.
The USP permits up to 2% of ordinary non-toxic 4. Gibson, W.; Powell-Evans, K. Validation Fundamentals;
impurities. However, impurities above 0.1% should Interpharm Press, Inc.: Buffalo Grove, IL, 1998.
5. Sucker, H., Ed.; Validation in Practice; Wissenchaftliche
be fully characterized and quantified. Impurities may Verlagsgesellschaft GmbH: Stuttgart, 1983.
include starting materials, by-products, intermediates, 6. Validation of Manufacturing Processes, Fourth European
degradation products, reagents, catalysts, heavy metals, Seminar on Quality Control, 1980.
7. FDA. Guideline on General Principles of Process Validation,
electrolytes, filter aids, and residual solvents. Known FDA, Rockville, MD, May 11, 1987.
toxic impurities must be held to a tighter standard, 8. FDA. Pre-approval Inspection/Investigations Guidance
i.e., below 0.1%. Manual, FDA, Rockville, MD, Oct. 1, 1990; 7346.832.
9. Fed. Reg. 43(190), Human and Veterinary Drugs, , 1978.
10. Lucisano, L.J.; Franz, R.M. FDA proposed guidance for
CMC changes: a review and industrial perspective. Pharm.
The ISO quality standards series Technol. 1995, 19 (5), 30–40.
11. WHO. Good Manufacturing Practices for Pharmaceutical
Products—Guidelines on the Validation of Manufacturing
The ISO 9000 Series was developed in 1987 by the Processes, WHO, Geneva, 1993.
International Organization for Standardization (ISO) 12. Skelly, J.P.; Scale-up of immediate-release oral solid
in Geneva, Switzerland. It is a comprehensive set of dosage. Forms. Pharm. Res. 1993, 10, 313–316.
13. Skelly, J.P.; Scale-up of oral extended-release dosage
management standards governing the operation of forms. Pharm. Res. 1993, 10, 1800–1804.
quality assurance to help develop and document a 14. Bolton, S. Process validation for hard gelatin capsules.
quality system that is useful for individual companies. Drug Cosm. Ind. 1984, 134 (3), 42–87.
15. Avallone, H.L. Development and scale-up of pharmaceuti-
The ISO 9000 Quality Management and Quality cals. Pharm. Eng. 1990, 10 (4), 38–41.
Assurance Standards, Guidelines for Selection and 16. FDA, Guide to Inspections of Oral Solid Dosage Forms Pre/
Use, provide basic definitions and concepts and Post Approval Issues for Development and Validation, FDA,
Rockville, MD, Jan 1994.
explain how to use the rest of the series (9001, 9002, 17. Berry, I.R. Process validation for soft gelatin capsules.
9003, and 9004). Drug Cosm. Ind. 1984, 134 (4), 26–35.
The ISO 9001 Quality System—Model for Quality 18. Nash, R.A. Process validation for solid dosage forms.
Pharm. Technol. 1979, 3 (6), 105–107.
Assurance in Design and Development provides for 19. von Doehren, P.J.; St. John, F.F.; Shively, C.D. An approach
quality assurance in the areas of design, installation, to the characterization and technology transfer of solid dosage
servicing, development, and production. It is useful form processes. Pharm. Technol. 1982, 6 (9), 139–156.
Unit–Validation

20. Sucker, H., Ed.; Validation in Practice; Wissenschaftliche

primarily for companies that design and develop their Verlagsgesellschaft GmbH: Stuttgart, 1983.
own products. 21. Berry, I.R., Nash, R.A., Eds.; Pharmaceutical Process Vali-
The ISO 9002 Quality System—Model for Quality dation, 2nd Ed.; Marcel Dekker, Inc.: New York, 1993;
Revised and Expanded.
Assurance in Production, Installation and Service 22. Chowhan, Z.T. Development of a new drug substance into
applies to manufacturers, distributors, and service ven- a compact tablet. Pharm. Technol. 1992, 16 (9), 58–67.
dors whose products have been designed and serviced 23. Fahrner, R. New role for pilot plants in product develop-
ment. Biopharm 1993, 6 (3), 34–37.
by a subcontractor. Such companies are exempt from
design control requirements. Both ISO 9001 and ISO
9002 are directly applicable to cGMPs. The connection
between the two independent systems is shown in BIBLIOGRAPHY
Table 5. Except for language, shades of meaning, and
stresses the documents are similar. Akers, J. Simplifying and improving process validation. J. Parent.
Sci. Technol. 1993, 47, 281–284.
The ISO 9003 Quality Systems—Model for Quality Avallone, H.L.; D’Eramo, P. Scale-up and validation of
Assurance in Inspection and Testing, designed for testing ANDA/NDA products. Pharm. Eng. 1992, 12 (6), 36–39.
laboratories and equipment distributors only requires Bala, G. An integrated approach to process validation. Pharm.
Eng. 1994, 14 (3), 57–64.
conformance to final inspection and testing procedures. Bolton, S. When is it appropriate to average and its relationship
The ISO 9004 Quality Management and Quality to the barr decision. Clin. Res. Reg. Affairs 1994, 11, 171–179.
Systems Elements—Guidelines provide standards and Chapman, K.G. A history of validation in the united states,
part I. Pharm. Technol. 1991, 15 (10), 82–96.
guidelines for quality management planning and Tomamichel, K.; Pharmaceutical quality assurance: basics of
implementation. validation. Swiss Pharma 1994, 16 (3), 13–23.
Veterinary Dosage Forms
J. Desmond Baggot
School of Veterinary Medicine, St. George’s University, Grenada, West Indies

INTRODUCTION containing ectoparasiticides for topical application to

various species, and darts containing sedative drugs for
Veterinary dosage forms are drug preparations capture and restraint of exotic animal species. The avoid-
designed for use in, or topical application to, one or ance of drug residues in tissues and animal products (milk
more species of domestic animal and/or other species and eggs) is a regulatory requirement of veterinary
of veterinary interest. Although the majority of veter- preparations administered to food-producing species.
inary dosage forms contain the same drugs as human
dosage forms, some veterinary preparations contain
drugs that are not widely used in humans. Examples VETERINARY DRUGS
include benzimidazole anthelmintics, macrolide endec-
tocides, salicylanilide flukicides, synthetic pyrethroids, Although the majority of drugs available as veterinary
chloramphenicol derivatives, a2-adrenoceptor agonists dosage forms were initially developed for use in
(sedative–analgesics), and antagonists. The converse humans, based on experimental findings in laboratory
applies to some classes of pharmacological agent animals, some drugs have been developed specifically
(e.g., benzodiazepine derivatives, tricyclic antidepres- for veterinary use. Anthelmintics and ectoparasiti-
sants) because of limited clinical indications for use cides/insecticides are primarily veterinary drugs. Some
in animals. Veterinary pharmacology differs from anthelmintics (e.g., ivermectin, mebendazole, albenda-
human pharmacology both in the diversity of species zole, pyrantel, piperazine, levamisole, praziquantel,
of interest and in the emphasis placed on the various bithionol) have been adopted for the treatment of
classes of drug. Some types of dosage forms are suit- parasitic infections in humans. Most ectoparasiticides
able for use in humans and certain animal species. are exclusively veterinary drugs. Some antimicrobial
They include parenteral solutions (although the agents within certain classes, which include sulfon-
concentration of the drug, the nature of the vehicle, amides, fluoroquinolones, macrolides, chloramphenicol
and other constituents of the preparation must be derivatives, and carboxylic ionophores, are available
considered); conventional tablets and capsules (amount only for use in animals. Relatively few pharmacologi-
of the drug relative to dosage requirement must be cal agents were initially developed as veterinary drugs,
considered); oral solutions and suspensions (pediatric although some drugs have indications for use in
preparations are generally appropriate for administra- animals that do not apply to humans (Table 1). Other
tion to small animals, especially cats); and conventional pharmacological agents have largely become veteri-
ophthalmic preparations. Sustained-release tablets and nary drugs by virtue of their clinical efficacy in animals
controlled-release transdermal drug-delivery systems and replacement by alternative drugs for use in
designed for use in humans could be used in medium humans. Examples include phenylbutazone, quinidine,
to large breeds of dog. It is because of the unique phenobarbital, and thiopental.

Veterinary–
physiological characteristics of each species, or closely Steroid (sex) hormones, gonadotrophins, gonado-

Virtual
related group of species, and the variation among spe- trophin-releasing hormones (synthetic forms and
cies in the dose–effect relationship of pharmacological GnRH analog), and synthetic prostaglandins (F2a type
agents that veterinary dosage forms of drugs are or analog) are used in female animals (cows, ewes,
required. Veterinary dosage forms designed specifically sows, and mares) to regulate various stages of the
for use in certain animals species include long-acting estrous cycle, whereas synthetic prostaglandins may
parenteral dosage forms of antimicrobial agents for be used to induce parturition. Melatonin, a modified-
intramuscular injection, oral pastes containing anti- release dosage form implanted subcutaneously behind
microbial agents or anthelmintics for horses, granules the ear of ewes, is used to stimulate the onset of cyclical
containing non-steroidal anti-inflammatory drugs or ovarian activity. Progestogens are used to synchronize
anthelmintics for addition to the feed for horses or pigs, estrus in groups of animals or to enable prediction of
respectively, modified-release ruminal boluses contain- the onset of estrus. On removal of the progestogen
ing anthelmintics for cattle or sheep, intramammary source (intravaginal device for cows and ewes; added
antimicrobial preparations for cows, preparations to feed for sows and mares), the negative feedback
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-100000378
Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved. 3941
 3942 Veterinary Dosage Forms

Table 1 Some pharmacological agents that are used exclusively in animals

Drug Classification Clinical indications
Xylazine a2, (a1)-Adrenoceptor agonist Sedation; analgesia preanesthetic medication
Yohimbine a2, (a1)-Adrenoceptor antagonist agonist Xylazine reversal
Detomidine a2-Adrenoceptor agonist Preanesthetic medication
Atipamezole a2, (a1)-Adrenoceptor antagonist Detomidine reversal
Acepromazine Phenothiazine-derivative tranquilizer Preanesthetic medication; sedation
Etorphine–Acepromazine Potent opioid agonist-PTZ-derivative Neuroleptanalgesia
tranquilizer
Diprenorphine Opioid antagonist Etorphine reversal
Droperidol–Fentanyl Butyrophenon m-opioid agonist Neuroleptanalgesia
Azeperone Butyrophenone Preanesthetic medication;
behavior modification (pigs)
Alfadolone–Alfaxalone Steroid anaesthetic (contraindicated Anesthesia
in dogs)
Flunixin Cyclo-oxygenase inhibitor Anti-inflammatory; analgesia; antipyresis
Metamizole (Dipyrone) Cyclo-oxygenase inhibitor (‘‘Similar’’ to other NSAIDs)

effect on the pituitary and hypothalamus is terminated, considered veterinary drugs. They include methyl-
and estrus is initiated. Progestogens (medroxyprogester- thioninium chloride (methylene blue), sodium nitrite,
one, megestrol, proligestone), generally administered by sodium thiosulfate, ammonium molybdate, and
subcutaneous injection, are used to suppress ovarian sodium calciumedetate. Phytomenadione (vitamin
activity (estrus) in dogs and cats, whereas altrenogest K1) is indicated for the treatment of warfarin and
(added to feed) is used to suppress estrus in cycling mares coumarin poisoning in animals. The substances are
or to synchronize estrus in gilts and sows. present in sweet vernal grass Anthoxanthum odoratum
Monensin, a carboxylic ionophore antibiotic, is and in spoiled sweet clover (Melilotus officinalis and
available as a premix (for addition to feed) used for M. alba) hay, and silage. Acetylcysteine and ascorbic
the prevention of coccidiosis caused by Eimeria spp. acid are used concurrently in the treatment of acetami-
in broiler chickens. Turkeys over 16 weeks of age, nophen (paracetamol) toxicity in cats. The acetylcys-
guinea fowl, and other game birds should not be given teine, which is administered by intravenous injection,
access to monensin-containing feed. Monensin is used serves as a precursor for glutathione replenishment,
as a production enhancer (improves feed conversion whereas ascorbic acid (administered intravenously
efficiency and growth rate) in beef cattle and dairy or orally) reduces methemoglobin to hemoglobin.
heifers up to the time of first service. It is available for Feline hemoglobin is particularly susceptible to oxidat-
use in cattle as a premix (sodium salt) for addition to ive damage (methemoglobinemia). Atropine sulfate is
the feed and as a modified-release ruminal bolus. The widely used in animals for preanesthetic medication
oral LD50 (mg/kg) of monensin differs among species: (although glycopyrronium is preferred in horses) and
horses, 2–3; sheep, 12; pigs, 16; cattle, 22; and chickens, at a much higher dose (25 to 40 times the preanesthetic
Veterinary–

200. Extreme care should be taken not to feed cattle, pig, dose, 44 mg/kg), often in conjunction with a cholin-
Virtual

or poultry rations or supplements to horses and to avoid esterase-reactivating agent (pralidoxime mesilate), in
accidental contamination in feedmills. Steroid hor- the treatment of organophosphate toxicity.
mone-growth promoters (bovine somatotropin, porcine This overview of veterinary drugs shows applica-
somatrotropin, bovine growth hormone-releasing fac- tions of these drugs in animals and how some aspects
tor) are available for use as production enhancers in of veterinary and human pharmacology differ in their
some countries, whereas in others (European Union orientation.
member states), their use is banned. The properties of
somatotropins and the dosage forms that have been
studied were comprehensively reviewed by Foster.[1] CATEGORIZATION OF SPECIES
The use of b2-adrenoceptor agonists such as clenbuterol
for production enhancement in cattle is illegal. Even though mammalian species differ in physical
Some antidotal substances used in the treatment characteristics (notably body weight) and behavior,
of plant or heavy metal toxicity in animals could be they possess the same body systems that perform
Veterinary Dosage Forms 3943

generally similar physiological functions. There are, coitus. Pharmacological intervention at any stage of
however, species-related adaptations that account for the reproductive cycle, whether it be to induce ovu-
the uniqueness of each species. The character of the lation in mares, ewes, or cows or to suppress estrus
adaptations underlies the feasibility of extrapolating or prevent ovum implantation in bitches or queens, is
scientific information obtained in certain species (such based on changing the plasma concentrations of the
as laboratory animals) to other species (domestic reproductive hormones that influence the particular
animals) and human beings. Because the pattern of process. To be successful, knowledge of the temporal
quantifiable adaptations can be generally described pattern of the various hormone concentrations in
mathematically, it is possible to make some predictions plasma is essential.
regarding interspecies extrapolation. The reliability of Because avian and mammalian species differ in
predictions on drug bioavailability and disposition many respects, these distinct classes of animal should
depends on knowledge of both the anatomical and be considered separately. Avian species that are
physiological similarities of and differences among ‘‘farmed’’ include chickens, turkeys, ducks, geese,
the species of interest. ostriches, guinea fowl, quail, and pheasants. The
The anatomical arrangement of the gastrointestinal term poultry refers to farmed domestic fowl that, in
tract serves as a basis for broadly categorizing common usage, includes chickens, turkeys, ducks and
domestic animals as ruminant species (cattle, sheep, and geese, whereas other farmed avian species are con-
goats) or monogastric species (horses, pigs, dogs, and sidered game birds. Application of the collective term
cats). Consideration of dietary habit somewhat refines poultry overlooks species differences in dosage require-
the categorization and enables the pattern of drug ments and drug residues in tissues as well as differences
absorption to be explained as well as the potential in susceptibility to toxicity of some drugs.
interaction among commensal microbial flora in the Birds (and reptiles) have a well-developed renal
digestive system and drugs, especially antimicrobial portal system that drains blood from the caudal por-
agents. Ruminant species are herbivores with a volu- tion of the body. Consequently, drugs administered
minous forestomach compartment in which microbial parenterally in the lower extremities (hind limbs) of
fermentation takes place continuously. Horses are those species pass through the kidneys before entering
monogastric herbivores with a small-capacity stomach the systemic circulation. This feature of blood flow to
and large-capacity colon where microbial digestion the kidneys provides the opportunity for first-pass
takes place. Dogs and cats are monogastric carnivores, excretion of water-soluble ionized drugs (e.g., b-lactam
whereas pigs are monogastric omnivores (similar to and aminoglycoside antibiotics) to occur.
humans) fed a vegetable diet. Because the urinary pH Fish, reptiles (which include crocodiles and alliga-
reaction is determined primarily by the composition tors), and amphibians are poikilothermic, i.e., cold-
of the diet, the usual pH range differs among herbivor- blooded animals. In contrast to homeothermic animals,
ous species (pH 7.2–8.4) and carnivorous species (pH the disposition of drugs in poikilothermic animals is
5.5–7.0), whereas in omnivorous species, urinary pH influenced by environmental temperature. When applied
can vary over a wide range (pH 4.5–8.2), but would to fish, the rate of drug elimination varies with the tem-
be expected to be alkaline in pigs, whereas it is usually perature of the water to which the fish are acclimatized.
acidic in humans. Urinary pH influences the extent of Studies of drug disposition in fish should generally be
reabsorption from the distal renal tubules and the carried out at more than one water temperature, and
half-life of weak organic acids and bases when a signifi- whether fresh water or sea water is contained in the tank
cant fraction (arbitrarily >20%) of the systemically depends on the species of fish. A complication arises in
available dose is eliminated by renal excretion. the case of salmon for example, because adult salmon

Veterinary–
The character of the female reproductive (estrous) live in sea water but spawn and grow as fingerlings in

Virtual
cycle varies with the animal species in several respects, fresh water.
that include duration of cycle, length of estrus (sexual
receptivity), and time of ovulation (Table 2).[2] In
seasonal breeding species (mare, ewe, doe, and queen), DOSAGE FORMS AND ROUTES OF
the time of year during which estrous cycles occur is ADMINISTRATION
strongly influenced by the photoperiod. The mare
and queen become anestrous in late autumn owing to The type of dosage form, the route of administration,
decreasing daylight hours, and cycles are re-established and site of injection of parenteral preparations depend
with increasing daylight in early spring. The converse on the animal species or group of related species
situation applies to ewes and does. The plane of (such as ruminant animals or poultry). The greatest
nutrition can affect the onset of estrous cycles in seaso- differences relate to oral dosage forms and topical
nal breeding species. The queen is unique among dom- preparations. Whether a veterinary dosage form is
estic animal species in that ovulation is induced by intended for individual animal treatment or for
 Virtual
Veterinary–
3944

Table 2 Average length of various stages of reproductive cycles of domestic animals

Time fertilized ova
enter uterus Time of implantation Type of Length of
Species Duration of estrus cycle Length of estrus Time of ovulation (after conception) (after conception) placenta pregnancy
Marea 21 days 5–6 days Last day of estrus 3–4 days 30–35 days Epitheliochorial 345 days
Cow 21 days 18 h 12 h after end of estrus 3–4 days 30–35 days Epitheliochorial 280 days
Ewea 17 days 36 h 30 h after beginning 3–5 days 15–18 days Syndesmochorial 147 days
of estrus
Doea (goat) 20 days 40 h 30–36 h after beginning 4 days 20–25 days Syndesmochorial 147 days
of estrus
Sow 21 days 45 h 36–40 h after beginning 3–4 days 14–20 days Epitheliochorial 113 days
of estrus
Bitch In estrus at 7–8 month Proestrus, 9 days; First or second day 5–6 days 15 days Endotheliochorial 64 days
intervals depending estrus, 7–9 days of estrus
on breed
Queena 16 days (non-bred) 5–6 days Induced 24–32 h 4 days 13 days Endotheliochorial 65 days
(pseudopregnancy after coitus
lasts 36 days)
a
Seasonally polyestrous.
(Modified from Ref.[2].)
Veterinary Dosage Forms
Veterinary Dosage Forms 3945

administration/application to a large group of animals high oral bioavailability, which implies reliable absorp-
(herd or flock medication) should be decided before the tion from the gastrointestinal tract and no more than
development of a dosage form. Convenience of admin- partial inactivation by the first-pass effect unless active
istration and cost of the drug preparation are foremost metabolites are formed, a half-life in the range of 4 to
considerations in determining the use of a veterinary 6 h, and a relatively high potency but reasonably wide
dosage form by animal owners. range of therapeutic plasma concentrations. Because of
variation between dogs and humans in the oral
bioavailability and the rate of elimination of most
GASTROINTESTINAL ABSORPTION lipid-soluble drugs, those that would be suitable for
formulating as sustained-release dosage forms often
The anatomical arrangement of the gastrointestinal differ between the two species. Theophylline meets
tract and associated digestive physiology govern the the criteria that make it suitable for formulating as a
pattern of drug absorption. In pigs, dogs, cats, and sustained-release oral preparation for administration
humans, the plasma concentration profile after the to dogs. Of the sustained-release oral dosage forms
administration of an oral solution or conventional that are commercially available, anhydrous theophyl-
(immediate-release) dosage form is generally similar line in tablet form (200 and 300 mg) is preferred for
in that it shows a reasonably well-defined single peak. use in dogs. This product has an oral bioavailability
Because the rate of gastric emptying differs among (theophylline) of 76%, and the dosage regimen
monogastric species and the anterior (upper) portion (20 mg/kg administered at12-h intervals) has been
of the small intestine is the principal site of absorption, predicted to maintain plasma concentrations within the
the time at which the peak plasma concentration therapeutic range (6–16 mg/ml) with less fluctuation
occurs may vary. An effective pH value of 5.3 in the in peak-to-trough theophylline concentrations than
microenvironment of the intestinal mucosal surface, other sustained-release dosage forms.[5] The dosage
rather than the reaction of intestinal contents (average regimen for the conventional dosage form (aminophyl-
pH 6.6), is consistent with observations on the absorp- line tablets) is 10 mg/kg administered at 8-h intervals
tion of drugs that are weak organic acids or bases. to dogs.
Under normal conditions, weak acids with pKa values Other drugs of veterinary interest that have been for-
above 3.0 and bases with pKa values below 7.8 are mulated as sustained-release oral dosage forms include
well-absorbed from the small intestine.[3] An alteration morphine, propranolol, quinidine, procainaminde, and
in the pH of the stomach or small intestine contents verapamil or diltiazem. Of these, sustained-release
can markedly change the degree of ionization of drugs morphine sulfate (tablets, 15 mg) has the greatest
that are weak organic electrolytes (acids or bases). At potential for use in dogs over 10 kg body weight. The
pH values below the pKa, weak acids exist primarily proposed dosage regimen, 1–2 mg/kg administered at
in the non-ionized form, which is the moiety that can 8- or 12-h dosage intervals, may be effective in the
readily be absorbed; the converse applies to weak management of chronic pain.[6] The sustained-release
bases. Lipid-soluble neutral molecules (digoxin, chlor- dosage form overcomes the shortcoming of conven-
amphenicol) and fluoroquinolones (amphoteric com- tional oral preparations of morphine, which is their
pounds) are well-absorbed in dogs, although the short duration of action in dogs. Phenytoin is a likely
systemic availability of norfloxacin is much lower than candidate drug for formulating as a sustained-release
that of enrofloxacin/ciprofloxacin or marbofloxacin. dosage form because it would enable phenytoin to be
Because of their polar nature, aminoglycoside antibio- used in dogs for the prevention/treatment of general-
tics are poorly absorbed from the gastrointestinal tract ized tonic–clonic seizures (grand mal epilepsy). The

Veterinary–
and must be administered parenterally in the treatment average oral bioavailability of phenytoin administered

Virtual
of systemic bacterial infections. as the conventional dosage form is 36%, the half-life in
After the administration of a sustained-release oral dogs is 3.5–4.5 h (dose-dependent), and the therapeutic
dosage form, the duration of drug availability for range of plasma concentrations is 10–20 mg/ml. Oral
absorption is limited by the sum of the residence times bioavailability of phenytoin in humans is 90%, and the
of the dosage form in the stomach and small intestine. apparent half-life, which is dose-dependent, is 15–24 h.
This has been estimated to be 9–12 h in dogs. In the The slower elimination of phenytoin in humans obviates
development of a sustained-release dosage form for the need for the development of a sustained-release
use in dogs, the aim is to provide an effective plasma dosage form.
concentration of the drug throughout the dosage inter- Even though the horse is a monogastric species, the
val (12 h) with an acceptable degree of fluctuation in stomach has a small capacity (8.5% of the gastrointes-
steady-state concentrations. The latter is determined tinal tract) compared with that of the pig (29%) and
both by the half-life of the drug and the dosage inter- the dog (62%). Expressed on the basis of volume
val.[4] Suitable candidate drugs should have reasonably capacity, the stomach of the horse, pig, and dog can
 3946 Veterinary Dosage Forms

hold 7–14, 5.5–7, and 3–8 L, respectively. The average fibrous material in feed.[7,8] The inadvertent contami-
pH value of gastric contents in horses is less acidic nation of horse feed with a feed additive premix
(pH 5.5; range, 4.5–6.0) than in other monogastric spe- approved for use as a production enhancer/growth
cies (pH 3–4), and a substantial portion of the stomach promotant in cattle or pigs, may cause toxicity, even
lining is composed of stratified squamous epithelium. death, in horses.
Under natural conditions of management, horses feed The anatomical arrangement of the gastrointestinal
continuously. The fibrous component of the feed is tractdistinguishes ruminant (cattle, sheep, and goats)
digested primarily in the large intestine, although from monogastric (horses, pigs, dogs, and cats) species
horses digest fiber less efficiently than do ruminant spe- (Table 3). The difference in digestive physiology
cies. Microbial digestion of fibrous carbohydrates to between the two groups of species determines the types
volatile fatty acids and break down of undigested diet- of oral dosage forms appropriate to administer and
ary protein to peptides and amino acids takes place in influences oral bioavailability of drugs. Susceptibility
the large intestine. The combined capacity of the to ingested plant toxicity differs between ruminant
caecum and colon occupies approximately 55% of the and monogastric species. The volume capacity of the
gastrointestinal tract, and the pH of large intestinal mature reticulorumen is 100–225 L in cattle and
contents is 6.6–6.8. Two unrelated features of the 10–25 L in sheep and goats and accounts for approxi-
digestive system are that horses do not possess a gall mately 60% of the total capacity of the gastrointestinal
bladder (similarly camelids and giraffes) and are tract. The forestomach contents vary from liquid to
unable to vomit. The temporal relationship between semisolid consistency, and the pH reaction is normally
feeding and oral dosing can greatly influence the maintained within the range 5.5 to 6.5 owing to copi-
pattern of drug absorption. Because the systemic ous secretion of alkaline saliva (pH 8.2–8.4) parti-
availability of most antimicrobial agents administered cularly rich in bicarbonate and phosphate buffers but
orally (paste formulations) or by nasogastric tube devoid of amylase. In addition to its buffering action,
(aqueous suspensions) to horses is significantly saliva has an antifoaming action that serves to prevent
decreased by feeding shortly before dosing, food dietary bloat. Salivary secretion, at a daily rate of
should be withheld for up to 2 h after administration 100–190 L in cattle and 6–16 L in sheep, is essential
of an antimicrobial agent. Feeding horses close to the for microbial digestion, which takes place continuously
time of administering phenylbutazone in various oral in the reticulorumen. Based on average values of saliva
dosage forms changed the pattern without altering flow and volume of the rumen liquid pool, the turnover
the extent of absorption of the non-steroidal anti- rate for reticuloruminal fluid was estimated to be
inflammatory drug.[7] The availability of a drug for 2.0/day for cattle and 1.1–2.2/day for sheep.[9] Despite
absorption from the small intestine of the horse, parti- the stratified squamous nature of its epithelial lining,
cularly when administered in conjunction with or the rumen has considerable absorptive capacity.[10,11]
shortly after feeding, may be limited by adsorption to Because absorption takes place by passive diffusion,
the ingested feed, especially hay. Under these circum- lipid-soluble drugs, whether neutral molecules or the
stances, absorption may occur in two phases, initially non-ionized form of weak organic acids or bases,
(1–2 h after drug administration) from the small intes- may be absorbed from the rumen. The theoretical equi-
tine and several (8–10) hours later from the large intes- librium distribution, expressed as concentration ratio,
tine (principal site) after microbial digestion of the of weak organic acids and bases of different pKa values
Veterinary–

Table 3 Relative capacity of components of digestive tract of domestic animal species

Virtual

Relative capacity (%) Relative capacity (%)

Component Horse Pig Dog Component (Sheep and Goat)

Stomach 8.5 29.2 62.3 Rumen 52.9
Small intestine 30.2 33.5 23.3 Reticulum 4.5
Cecum 15.9 5.6 1.3 Omasum 2.0
Large colon 38.4 Abomasum 7.5
31.7 13.1
Small colon and rectum 7.0 Small intestine 20.4
Cecum 2.3
Colon and rectum 10.4
Veterinary Dosage Forms 3947

between saliva (pH 8.2) or ruminal contents (pH range particles of reticuloruminal contents is ‘‘pumped’’ by
5.5–6.5) and plasma (pH 7.4) is presented graphically the omasum (third compartment of the forestomach)
(Fig. 1).[12] into the abomasum (true stomach). During the two-
Ruminal micro-organisms are capable of at least stage transfer process, water and electrolytes are
partially metabolizing some drugs (e.g., trimethoprim, absorbed, and the size of particulate matter in the
chloramphenicol, nitroxynil, digitalis glycosides) either digesta is reduced. The abomasum, which accounts
by hydrolysis or reduction, which would decrease the for approximately 4–5% of the capacity of the gastro-
amount of drug available for absorption. Intraruminal intestinal tract in adult cattle and 7.5% of gastro-
biotransformation makes ruminant species susceptible intestinal capacity in sheep and goats, is the only
to toxicity caused by ingestion of plants containing compartment of the ruminant stomach that secretes
cyanogenetic glycosides (cyanide poisoning) or accu- digestive juices. Secretions from the fundic area of
mulated nitrate (nitrite poisoning). Overuse of the abomasum contain hydrochloric acid, pepsin
nitrogenous fertilizers contributes to the incidence of and, in suckling preruminant animals, rennin (a milk-
nitrite poisoning. Antidotal substances that are used coagulating enzyme). Mucus is produced by the colum-
(injected intravenously) to treat these toxicities are nar epithelial cells that line the abomasum. The pH
sodium nitrite, followed by sodium thiosulfate for reaction of abomasal contents does not vary much
cyanide poisoning and methylthioninium chloride and is usually close to pH 3.0.[14]
(methylene blue) for nitrite poisoning. In ruminant Because of the large volume of reticuloruminal con-
species, in which chemical compounds can be altered tents, a drug can attain only a low concentration in the
(activated or detoxified) by microbial action in the reticulorumen, whether administered in solution, sus-
forestomach and in which the whole physiological pension, or solid dosage form. In dissolution of solid
tempo of the body is so dependent on ruminal activity, dosage forms, dilution in the large volume of fluid
no pharmacological or toxicological investigation can and binding to particulate matter would decrease the
be interpreted without full consideration of the basic rate, but not necessarily the extent, of drug absorption.
diet and feeding regimen.[13] Lipid-soluble neutral molecules and the non-ionized
After comminution by rechewing and microbial form of weak organic electrolytes, particularly organic
digestion, the liquid component with suspended acids, should normally be well-absorbed from the reti-
culorumen. When aspirin (pKa 3.5) in a solid dosage
form (60 g, which is equivalent to 3.9 g of oral bolus)
Selection of dosage form was administered to cows, salicylate was slowly
100
absorbed, and systemic availability was 50–70%.[15]
pH 5.5
The 12-h dosage interval for aspirin in adult cattle is
based on the rate of absorption rather than on the
Rumen, base half-life of salicylate, which is 0.8 h.
10 Saliva, acid Benzimidazole anthelmintics (albendazole, fenben-
pH 6.5
dazole, oxfendazole, which is fenbendazole sulphox-
Concentration ratio

ide); probenzimidazoles (netobimin and febantel,

which are metabolically converted to albendazole and
1 fenbendazole, respectively); and salicylanilide fluki-
cides (closantel, rafoxanide, oxyclozanide) are adminis-
Saliva, base
tered as oral suspensions to ruminant animals. For
pH 6.5 anthelmintic drugs oral suspensions have an advantage

Veterinary–
0.1 over oral solutions in that the delayed availability of

Virtual
Rumen, acid drug for absorption extends the duration of action.
Reducing the level of feed intake (for 36 h before and
36 h after dosing) delays the onward passage of
0.01 pH 5.5
ruminal fluid with suspended matter from the reticu-
4 5 6 7 8 9 10 lorumen to the absomasum and small intestine and
pKa would allow more time for drug dissolution and
absorption to occur. When the rate of onward passage
Fig. 1 Expected equilibrium distribution between saliva or
of ruminal fluid was decreased, by temporarily reduc-
rumen contents and plasma of acids and bases of differing
pKa. Concentration ratio is the ratio of the salivary or rum- ing feed intake, in sheep, the systemic availability of
inal concentration to concentration free in the plasma, calcu- oxfendazole was increased.[16] The systemic availability
lated separately for acids and bases, for saliva of pH 8.2 and of rafoxanide and triclabendazole, administered as
rumen contents over a range of pH 5.5–6.5, assuming plasma oral suspensions, was higher in housed lambs fed hay
is pH 7.4. (From Ref.[12].) and concentrates than in lambs grazing on pasture.[17]
 3948 Veterinary Dosage Forms

The effect of dietary regimen/composition on systemic systems designed for use in humans is limited by gastric
availability of the flukicides could be attributed to emptying time, which can be up to 12 h.[19]
the time allowed for dissolution and absorption of
the drugs.
Closure of the reticular groove would allow an oral FIRST-PASS EFFECT
solution to flow directly from the cardiac orifice of the
rumen into the absomasum, bypassing the rumen. Having traversed the gastrointestinal mucosal barrier,
Spontaneous closure of the groove occurs reflexly in drug molecules are conveyed in hepatic portal blood
some animals although at least partial closure can be to the liver, where they are subjected to the first-pass
chemically induced. Should rapid absorption of a drug effect before entering the systemic circulation. The
(such as a non-steroidal anti-inflammatory drug) from first-pass effect applies to all animal species and, owing
the gastrointenstinal tract be desired, an oral solution to the generally higher capacity of the liver of herbivor-
of the drug could be administered immediately after ous species (ruminant animals and horses) to meta-
inducing closure of the reticular groove. This can be bolize lipid-soluble drugs by microsomal oxidative
achieved generally in cattle by administering orally a reactions, is likely to decrease the systemic availability
solution containing sodium bicarbonate and in sheep of these drugs to a greater extent in herbivorous than
by administering orally a copper sulfate solution or in non-herbivorous species(dogs, cats, pigs, humans).
injecting intravenously a dose (0.3 U/kg) of lysine– In ruminant species (cattle, sheep, goats) and Equidae
vasopressin. Closure of the reticular groove would be (horses, ponies, donkeys), triclabendazole, adminis-
unwanted in the case of an oral liquid, such as dimeti- tered as an oral suspension, is converted by hepatic
cone emulsion or poloxaline, used to treat frothy bloat first-pass metabolism to triclabendazole sulfoxide
in cattle. In young ruminant animals, reflex closure of (active metabolite), which is subsequently converted
the reticular groove, induced by suckling, allows milk to the sulfone (inactive) metabolite. In many species,
to flow directly to the abomasum. the first-pass effect can substantially reduce the sys-
Advantage can be taken of the anatomical arrange- temic availability of orally administered lipid-soluble
ment of the forestomach of ruminant animals by drugs that undergo extensive biotransformation in
administering a modified-release ruminal bolus that the liver (e.g., diazepam, propranolol, verapamil).
will remain lodged in the reticulorumen for a pro- Although presystemic metabolism occurs primarily in
longed period. Slow-release ruminal boluses containing the liver, it can also take place in the small intestinal
trace elements (cobalt oxide, copper oxide, cobalt and mucosa during the absorption process. Some drugs
copper in sodium phosphate glass matrix, and sele- that show incomplete systemic availability after oral
nium as sodium selenate) are commercially available administration to dogs are listed in Table 4. Presyste-
for administration to cattle or sheep, and the com- mic metabolism in the liver is responsible for incom-
pound ruminal bolus can be administered to farmed plete systemic availability of the majority of these
adult deer. Controlled-release ruminal boluses contain- drugs. The oral bioavailability of digoxin was
ing certain anthelmintic drugs (ivermectin, fenbenda- enhanced in humans when the drug was administered
zole, oxfendazole, morantel) or the production as an aqueous alcoholic solution in gelatin capsules
enhancer monensin are available for oral administra- rather than in tablet form, even though the tablets
tion, using a specialized delivery device, to beef cattle had a satisfactory dissolution rate.[20–22] In a similar
within a specified range of body weight (100–400 kg). manner, increased relative bioavailability, based on
These systems are designed either to continuously comparison of areas under the plasma concentration–
release the drug into ruminal fluid for a prolonged time curves of flufenamic acid, was achieved in dogs
Veterinary–

period (generally 120–140 days) or to intermittently when the drug was administered in soft rather than
Virtual

release pulse doses (oxfendazole bolus) at a predeter- hard gelatin capsules; the average increase was
mined interval (approximately 3 weeks). This interval 34%.[23] In both instances, the higher oral bioavailabil-
generally coincides with the prepatent period of ity was attributed to physicochemical factors brought
the major gastrointestinal tristrongylids of cattle.[18] into play by adjuvants in the soft gelatin capsules.
Controlled-release ruminal boluses that continuously The influence that formulation, or rather dosage form,
release ivermectin, fenbendazole, or morantel into can have on the systemic availability of an orally admi-
ruminal fluid are available for use in beef cattle, nistered drug was shown in horses given racemic
whereas controlled-release boluses that deliver alben- ketoprofen (2.2 mg/kg). The systemic availability of
dazole or ivermectin over a period of 100 days are the S(þ)- and R(
)-enantiomers was 54.2 and 50.5%,
available for use in sheep (35–65 kg in body weight). respectively, after administration of micronized
The delivery systems are retained in the reticulorumen racemic ketoprofen powder in hard gelatin capsules
at least throughout the entire period of drug release. to horses with restricted access to feed. When ketopro-
The retention of oral controlled-release drug delivery fen powder from the same batch was administered as
Veterinary Dosage Forms 3949

Table 4 Systemic availability of some orally administered drugs in dogs

Dose Systemic
Drugs (dosage form) (mg/kg) availability (%) Site of metabolism/other factor
Phenobarbitone (tablet) 10 86–96 Liver
Valproic acid (tablet) 40 78 Liver
Phenytoin (tablet) 15 36a Liver
Salicylate (aspirin tablet) 250 mg total 45 Liver
Ibuprofen (gelatin capsule) 5 60–86 Liver
Naproxenb (gelatin capsule) 5 68–100 —
Theophylline (conventional aminophylline tablet) 10 91 Liver
Diazepam (tablet) 2 1–3 86c Liver þ (intestinal mucosa)
Lidocaine (solution) 10 15 Liver
Procainamide (tablet) 25 85a,d
Propranolol (conventional tablet) 80 mg total 2–17 Liver
Verapamil 0.5 15 Liver
Digoxin (Lanoxin tablet) 1 mg total 80 Dissolution
Cephalexin monohydrate (capsule) 20 57 Dissolution
Norfloxacin (tablet) 5 35a Dissolution þ liver
Enrofloxacin (tablet) 5 100e —
f
Allopurinol (tablet) 15 70 Intestine þ liver (xanthine oxidase)
a
Average oral bioavailability; wide individual dog variation.
b
Naproxen has an unusually long half-life in dogs.
c
Total active benzodiazepine.
d
Dogs do not form N-acetylprocainamide (active metabolite).
e
Total antimicrobial active fluoroquinolone.
f
Average oral bioavailability is 14% in the horse.

an oil-based paste, systemic availability of the S(þ)- unlike in the humans and dog, appears to be substan-
and R(
)-enantiomers was 5.75 and 2.7%, respectively, tially into the hepatic portal vein, minimal avoidance
regardless of the feeding schedule.[24] To avoid hepatic of hepatic first-pass metabolism of drugs absorbed
first-pass metabolism, glyceryl trinitrate (nitroglycerin) from the rectum could be anticipated in horses.
is formulated in a variety of dosage forms, which
include parenteral solution, spray for sublingual
application, sustained-release tablet, and transdermal ORAL DOSAGE FORMS
therapeutic systems, for use in humans, and as an
ointment for topical application to dogs (cardiogenic The oral route of drug administration is safer than par-
pulmonary edema) or horses (acute laminitis). Sub- enteral routes and avoids tissue irritation at injection
lingual administration avoids the first-pass effect, but sites. However, wide variation, both inter- and intra-
it is not feasible to administer solid dosage forms by species, in systemic availability is a feature of orally

Veterinary–
this route to animals. administered drugs. The convenience of oral drug

Virtual
Although the first-pass effect is a major source of administration depends on the species of animal and
species variation in systemic availability of orally the dosage form of the drug. To facilitate drug admin-
administered drugs that undergo extensive hepatic istration and take into account the anatomy and physi-
metabolism, another important source of variation is ology of the digestive system and the average body
metabolism by ruminal microorganisms. Some drugs weight of the various animal species, the requirements
(nitroxynil, chloramphenicol, digitalis glycosides) are of oral dosage forms differ among species. Consider-
metabolized in the rumen to such an extent that par- ation must be given to size of the total dose (amount
enteral administration is required for clinical efficacy. of drug) to be administered and oral bioavailability
An advantage of rectal over oral administration of in the animal species.
lipid-soluble drugs is partial avoidance of the first-pass Oral dosage forms available for administration to
effect. The extent to which the first-pass effect is animals include oral solutions, liquids, suspensions,
avoided appears to be less in dogs than in humans. gels, pastes, capsules, tablets, ruminal boluses, powders
Because venous drainage of the rectum of the horse, and granules for addition to feed, soluble powders for
 3950 Veterinary Dosage Forms

addition to drinking water or fish medicating baths, metabolism by ruminal micro-organisms and/or
and premixes for addition to feed for livestock or chemical degradation in the ruminal environment.
poultry. The type of dosage form is determined by Encapsulation of insect development inhibitors (e.g.,
the solubility and physicochemical properties of the diflubenzuron) or insect growth regulators (e.g., meth-
drug, the species of animal for which the dosage form oprene) has potential application in cattle. These
is intended, and whether a reasonably rapid onset or a substances interfere with the metamorphosis and
prolonged duration of effect is required. Liquid dosage reproductive capacity of arthropod parasites, many
forms (oral solutions and liquids) provide readily avail- of which breed in cattle dung. Dichlorvos, an organo-
able drug for absorption, particularly in monogastric phosphorus compound with activity against gastro-
species. Oral liquids, such as dimeticone emulsion intestinal nematodes including whipworms (Trichuris
and poloxaline (anionic surfactant), are used for the spp.), is incorporated into polyvinyl chloride resin
treatment of frothy bloat in cattle. An oral liquid pellets (plasticization) for addition to the feed for pigs
containing propylene glycol (glucose precursor) for or canned food for dogs. This dosage form protects
addition to drinking water or for preparation of an dichlorvos from hydrolytic degradation and provides
oral solution is indicated for adjunctive treatment of slow release into the gastrointestinal tract, which
ketosis in cattle and sheep. Aqueous suspensions are greatly increases the margin of safety of the drug.
administered by nasogastric tube to horses, by mouth A sustained-release dosage form provides an initial
(as a drench) to ruminant animals and by mouth amount of drug sufficient to provide a desired thera-
(sometimes with the aid of an oral syringe) to dogs peutic concentration and continuously releases the
and cats. Oral pastes and gels are semisolid dosage drug at a constant (zero-order) rate for an extended
forms supplied in preloaded calibrated syringes period. The margin of safety of the drug is an impor-
designed for convenience of drug administration to tant consideration, because sustained-release dosage
horses by their owners. The formulation of a paste forms contain a large amount of drug and the potential
must be such that it is syringeable over a wide range exists for ‘‘dose-dumping’’ with resultant toxicity. For
of ambient temperatures, is moderately tenacious so drugs with half-lives in the range of 4 to 6 h, sustained-
that it will adhere to the tongue, and is tasteless or release dosage forms could provide a dosage interval
suitably flavored (mint or apple flavor appears to of 12 h in dogs or 24 h in horses. Very few sustained-
be preferred by horses). Classes of drugs that could release preparations have been developed for use in
be formulated as oral pastes or gels for administration either species. This type of dosage form has to be
to horses include anthelmintics, non-steroidal anti- swallowed intact.
inflammatory drugs, and some antimicrobial agents. Controlled-release ruminal boluses are designed
Application of an anthelmintic paste, at the appropri- either to continuously release drug (an anthelmintic
ate dose of the drug, to the distal forelimbs of cats is a or production enhancer) at a constant rate for a pro-
convenient alternative to oral administration of a tab- longed specified period or to intermittently deliver
let or capsule. Solid dosage forms must undergo dis- pulse (usually five) doses at predetermined intervals.
integration and dissolution in the stomach before Each product is designed for use in either cattle or
absorption of the drug can occur. Drug release from sheep within a specified range of body weight. Reten-
solid dosage forms delays the rate of absorption com- tion of orally administered controlled-release systems
pared with that from an oral solution. With regard to in the reticulorumen is dependent on either density or
the quality of solid dosage forms, capsules offer an geometry of the system. The ivermectin (Ivomec)
advantage over tablets in that the particle size and dis- ruminal bolus, designed for use in cattle between 100
tribution of the active ingredient are rarely altered by and 400 kg body weight, contains 1.72 g of ivermectin
Veterinary–

the capsule-filling process, whereas physical stresses that is continuously released into ruminal fluid at a
Virtual

(compression and heat) are imposed in the manufac- constant rate (12.5 mg/day) over a period of 135 days.
turing process of tablets. In addition, capsules provide The bolus is a cylindrical device that consists of an
more rapid dissolution than do conventional tables of outer semipermeable membrane enclosing a metal den-
the same active ingredient. The pattern of release of sity element at one end, an osmotic energy source at
coated pellets or granules is more predictable when the other end, and a formulation containing the drug
packed in capsules than compressed into tablets. in the center. The osmotic energy source is composed
Capsules, however, are generally more expensive than of a tablet containing a polymeric salt mixture.
tablets containing the same drug. Because of the higher Absorption of water through the semipermeable
manufacturing cost, soft gelatin capsules should membrane causes expansion of the tablet, which drives
probably be used in preference to hard gelatin capsules the ivermectin wax formulation through the exit port
only when the fill is liquid or bioavailability of the (channel) in the density element[25] (Fig. 2A). The
drug is significantly superior. An encapsulated active fenbendazole ruminal bolus (Panacur), designed for
ingredient, until released, would be protected against use in cattle between 100 and 300 kg body weight,
Veterinary Dosage Forms 3951

A a period of at least 90 days. There is no withdrawal

Dosage formulations period associated with the use of the morantel tartrate
Exit port trilaminate bolus because the combination of poor
absorption and first-pass hepatic metabolism makes
systemic availability of morantel exceedingly low.
Exit passageway The Captec, also known as the Laby,[27] device is a
controlled-release ruminal capsule designed for use in
Densifier
sheep within a specified range of body weight that
continuously releases the anthelmintic contained
within, either albendazole (32.5 mg/day) or ivermectin
Drug+vehicle
(1.6 mg/day), over a period of 100 days. The device
consists of a polypropylene barrel with a spring-loaded
Injection molded
plunger at the closed (top) end and an orifice (series of
semipermeable perforations) at the base (Fig. 2B). A pair of poly-
membrane propylene wings attached to the top of the barrel is
secured in a folded position (compressed configur-
ation) by water-soluble tape to enable oral administra-
Osmotic driving
member (tablet) tion. The barrel is loaded with tablets containing the
anthelmintic, either albendazole (3.85 g) or ivermectin
(160 mg). After administration of the ruminal capsule,
the tape securing the pair of wings dissolves, and the
wings open to a predetermined angle (expanded configur-
ation) that prevents both regurgitation and reticulo-
B
omasal passage. The rate of drug release is determined
by a combination of factors that include the formulation
28mm
of the tablets, the diameter of the orifice, and the pressure
exerted on the tablets by the plunger. The ruminal fluid
Plunger bathing the orifice of the device brings about dissolution
X. Plunger movement measurement
(Initial value 94-96mm; of the tablets.
the value decreases as The modified-release ruminal bolus containing
7 Tablets plunger movement increases)
containing oxfendazole, designed for use in cattle in the body
albendazole weight range of 200–400 kg, delivers pulse doses
(1.25 g) at 3-week intervals over a period of 105
days.[18] The bolus containing five tablets (Autoworm 5)
End view of Captec device releases the first dose at 21 days after administration
whereas the bolus containing six tablets (Autoworm
Fig. 2 (A) IVOMEC schematic; (B) captec schematic.
6) releases the first dose on the day of administration.
Drugs in powder form or prepared granules can be
added to the feed for pigs. Shortcomings associated
contains 12 g of the anthelmintic which, is continu- with this method of administration include uncertainty
ously released into ruminal fluid over a period of up as to the amount of drug (dose) ingested and inter-
to 140 days. Release of fenbendazole is controlled by animal variation in oral bioavailability. The drug must

Veterinary–
the erosion of two magnesium alloy tubes; each tube have a wide margin of safety and be palatable in the

Virtual
contains five tight-fitting cylindrical tablets (6 g of feed and, importantly, the animal must be feeding.
fenbendazole). The magnesium alloy tubes are pro- Inappetence or indifference to feeding is a usual feature
tected from the ruminal contents by a series of close- of illness in animals. Addition to feed, often referred
fitting rigid plastic rings; the central ring connects to as top dressing of feed, is generally an unreliable
both tubes. The bolus is completely biodegradable method of drug administration to horses. However,
and leaves no residual device in the forestomach.[26] the packaging of powders or granules in unit dose
The morantel tartrate (Paratect Flex) ruminal bolus, sachets provides convenience of drug administration
designed for use in cattle over 100 kg body weight, for owners when the number of animals to be dosed
contains 11.8 g of morantel base. The bolus consists is small. Unit dose sachets must be prepared on an
of a drug-impregnated polymer packaged in a cylindri- individual species basis because the dose level (mg/kg)
cal trilaminate cartridge. Release of the drug (approxi- may differ among species, and the average body weight
mately 150 mg/day) takes place by diffusion through range of domestic animal species differs by approxi-
the semipermeable membrane into ruminal fluid over mately 200-fold. Soluble salts of drugs in prepared
 3952 Veterinary Dosage Forms

solutions or powder form can be administered to larger in diameter (40–80 Å) than those in the intestinal
poultry and other farmed avian species by addition epithelium. Polar drugs such as aminoglycoside anti-
to drinking water or to fish by addition to the water biotics are rapidly and completely absorbed from
in a medicating bath of known volume capacity. intramuscular injection sites, whereas they are very
Sachets containing various trace elements (cobalt, poorly absorbed from the gastrointenstinal tract when
copper, iodine, selenium) for addition to drinking water the epithelial membrane is intact. Drug absorption
delivered by dispenser are available for the correction from intramuscular injection sites is generally assumed
of mineral deficiencies in cattle. The mineral content to be a first-order process. However, this assumption
of each sachet suffices for 25 cattle for a period of 7 may not be entirely valid, particularly during the initial
days. Several factors to consider in the design of premix stage when absorption may obey zero-order (non-lin-
formulations and to ensure their satisfactory mixing ear) kinetics. Local and systemic factors that influence
in bulk feed are addressed in detail by Klink et al.[28] the rate of absorption seldom remain constant
Premixes must always be diluted to the approved use throughout the absorption process. Increased blood
level, usually parts per million (ppm) [g/tonne (feed) flow to skeletal muscle at the site of injection promotes
or mg/L (water)], for the animal species. absorption, whereas the administration of a drug that
causes local vasoconstriction or the presence of a
disease state that decreases skeletal muscle perfusion
INTRAMUSCULAR INJECTION delays absorption. Deposition of the injected prep-
aration between muscle masses (intermuscular) or in
Intramuscular injection is a commonly used route for adipose tissue and the injection of a preparation that
drug administration in animals. This route offers con- causes tissue irritation or precipitation of the drug
venience in that it is easy to apply and, because a size- at the injection site produce an erratic pattern of
able proportion of veterinary parenteral preparations absorption that is reflected in the plasma concentration
are formulated as long-acting dosage forms, either a profile of the drug.
single dose may suffice or a long dosage interval can The site of intramuscular injection can affect the
be used. There are, however, a number of factors to plasma concentration profile and bioavailability of a
consider before choosing this route of administration. drug, particularly when a long-acting preparation is
They include the purpose for administering the drug, administered. The variation in the pattern of absorp-
the acceptable time interval between drug adminis- tion can be attributed to regional differences in blood
tration and onset of the desired effect, the suitability flow to skeletal muscles and in absorptive surface area.
of available parenteral preparations for use in the In cattle and goats, intramuscular injection in the
particular animal species, the withdrawal period(s) in lateral neck provides superior absorption to injection
food-producing animals, and the cost of the drug in the buttock (M. semitendineus) and thigh muscle
preparation for the anticipated course of treatment. (M. quadriceps femoris), respectively, or subcutaneous
The suitability of a parenteral preparation for use in injection in the lateral neck. In pigs, the lateral neck
a particular species primarily depends on the formu- should always be used as the site for intramuscular
lation of the preparation. Significant formulation injection of parenteral preparations. The reasons for
variables include the concentration of drug in a prep- selecting this site include better absorption than in
aration, which determines the volume to be injected, other sites, less residual drug at the injection site, and
the nature of the vehicle and other ingredients (solu- avoidance of damage to the carcass. In dogs and cats,
bilizers, preservatives), and the likelihood of causing the quadriceps muscle mass is the preferred site for
irritation at the injection site. The horse is the least intramuscular injection, which should be performed
Veterinary–

tolerant of domestic animal species to injection-site slowly, of parenteral preparations. After the injection
Virtual

irritation, and drugs in oily vehicles should never be of procaine penicillin G (20,000 IU/kg) at various
administered by injection to horses. intramuscular sites and subcutaneously in the cranial
Whereas the formulation of a preparation deter- part of the pectoral area in horses, the peak plasma
mines the pattern of absorption and influences sys- concentration and systemic availability of penicillin
temic availability of the drug, the volume of the G were highest when the long-acting preparation was
preparation deposited in muscle and the vascularity injected i.m. in the neck region (M. serratus ventralis
of the injection site determines the rate of absorption. cervicis). This was followed, in descending order
The barrier to absorption is the capillary endothelium, of injection site, by M. biceps > M. pectoralis >
which most drugs readily penetrate by passive diffusion, M. gluteus, or subcutaneously (Fig. 3).[29] The intra-
although small water-soluble molecules may enter capil- muscular injection technique must ensure that inadver-
laries by bulk flow through intercellular ‘‘pores’’ in the tent intravenous administration does not occur.
endothelial membrane. Aqueous channels (pores) in Potential disadvantages of intramuscular injection
the capillary endothelium are approximately 10-fold are the deposition of the preparation in adipose tissue
Veterinary Dosage Forms 3953

4 system, first-pass renal excretion may decrease the

systemic availability of drugs that are primarily elimi-
nated by the kidneys (e.g., b-lactam and amino-
Plasma concentration (IU/ml)
M. serratus
3 M. biceps glycoside antibiotics) when injected intramuscularly in
M. pectoralis
M. gluteus
the thigh of birds or caudal half of the body of reptiles.
Subcutaneous

2
SUBCUTANEOUS INJECTION

1 Subcutaneous injection is an alternative route to intra-

muscular injection for administration of parenteral
preparations to domestic animals. This route of admin-
0 istration is used most often in dogs and cats and only
0 5 10 24 occasionally in horses. Most of the factors that influ-
Hours after administration ence drug absorption from skeletal muscle also apply
to subcutaneous sites. They include the concentration
Fig. 3 Mean plasma penicillin concentration–time curves of drug in the parenteral preparation, the nature of
after 20,000 IU of procaine penicillin g/kg was administered
the vehicle and other components of the formulation,
to 5 animals (4 horses, 1 pony) at 5 different sites. (From
the total volume of the preparation administered,
Ref.[29].)
blood supply to subcutaneous tissue and area of the
absorptive surface, tissue irritation, precipation of drug
or intermuscular fascial planes and the production of at the site of injection, and persistence of drug residues.
tissue damage with persistence of drug residue at the Absorption from subcutaneous sites is often slower
site of injection.[30–32] Tissue damage is more likely to and more erratic than from intramuscular sites because
be caused by constituents of the formulation than the of the limited and more variable blood flow to sub-
drug substance per se. Useful antemortem methods cutaneous tissue. However, the absorptive area may
of assessing the extent of tissue irritation and the rate be larger and can be expanded by massaging the skin
of resolution at an intramuscular injection site include covering the region of the injection site. The degree
the echographical examination of the muscle tissue of tissue irritation caused by some parenteral prepara-
in the immediate vicinity of the injection site[33] and tions appears to be greater after subcutaneous than
the monitoring of plasma creatine kinase (CK) intramuscular injection.[36,37] Because the damaged
activity.[34,35] These methods have the distinct advan- tissue surrounding a subcutaneous injection site can
tage of being applicable to the live animal. Should be trimmed from the carcass of a slaughtered animal,
moderate to severe tissue damage be evident, the extent the likelihood of violative drug residues in the meat
of the damage and precise nature of the lesion can be is reduced. Subcutaneous injection is the preferred
described on postmortem examination. The use of route of administration for some poorly soluble sus-
some tissue-damaging parenteral preparations in pensions. When formulating sustained-release par-
food-producing animals is unavoidable, and the speci- enteral preparations, a compromise may have to be
fied withdrawal period(s) must be applied. The with- made between the rate of drug release and the degree
drawal period for a drug varies with the formulation of tissue irritation caused by the preparation. Various
of the dosage form (preparation), which should be modified-release devices for subcutaneous implan-
administered only by the recommended route and tation have been developed as an alternative to injec-

Veterinary–
may differ between animal species. Fish are ectother- tion of sustained-release oily suspensions. Devices

Virtual
mic and their basal metabolic rate, which markedly developed include an implantable osmotic pump that
influences the rate of elimination of drugs, varies with delivers growth hormone-releasing factor over a period
the temperature of the water to which they are acclima- of 7 days in steers and wethers.[38] Another subdermal
tized. It follows that withdrawal periods, stated in implant is a diffusion polymeric matrix system that
degree days, for drugs used in ‘‘farmed’’ fish (primarily releases estradiol at an approximately zero-order rate.
by addition to feed or bath water) will vary with ambi- A modified-release melatonin implant designed to
ent water temperature. advance the time of onset of cyclical ovarian activity
In cats, puppies, and piglets, particular attention in ewes is commercially available. Although great
should be given to the drug concentration in paren- efforts have been made toward technological develop-
teralpreparations and, when giving an intramuscular ment of modified-release devices for subcutaneous
injection in the thigh (particularly of cats), to avoid implantation in cattle and sheep, the use of implants
causing damage to the sciatic nerve. Because avian in food-producing animals does not appear to have
and reptilian species have a well-developed renal portal gained wide acceptance over the past decade.
 3954 Veterinary Dosage Forms

INTRAVENOUS INJECTION

Concentration of amikacin in plasma (µg/ml)

50

When a parenteral drug solution is administered intra-

20
venously, it is assumed that the total dose of the drug
is completely available systemically. This assumption Intravenous
10
overlooks the fact that an intravenously administered Intraosseous
drug passes through the lungs, which constitute an
5
organ of biotransformation, before being distributed
throughout the body. Intravenous injection produces
a prompt pharmacological response and overcomes 2
the variability in absorption associated with other
routes of drug administration. Parenterals, excluding 1
long-acting preparations, that are too irritant to be
injected by other routes may be cautiously adminis- 0.5
tered intravenously. It must be ensured that perivascu- 0 1 2 3 4 5 6 7 8 9 10
lar leakage at the site of injection does not occur. Time (h)
Intravenous injection of irritant solutions can cause Fig. 4 Concentrations (mean  S.D.) of amikacin in
thrombophlebitis. Once a drug has entered the sys- plasma after i.v. and i.o. administration of amikacin sulphate
temic circulation, its removal from the body is entirely (7 mg/kg body weight) to 6 foals at 3 to 5 days of age.
dependent on the elimination (biotransformation and
excretion) processes. Pharmacokinetic parameters that
describe the disposition (distribution and elimination) complete systemic availability of the drug and allows
of drugs are based on the plasma concentration pro- greater control over the intensity (magnitude) of the
files after intravenous injection of single doses. effects produced. The relatively short dosage interval
Assumptions made in the estimation of absolute bio- associated with multiple dosing is the principal disad-
availability, which is based on comparison of total vantage of the use of intravenous injection in animals.
areas under the plasma concentration–time curves Long-acting preparations administered by intramuscu-
after extravascular and intravenous administration lar injection constitute the parenteral dosage form
of the drug, are that the intravenously administered most widely used in farm animals.
dose is completely available systemically and that the Intraosseous administration is a feasible alternative
clearance of the drug is not changed by the route of to intravenous injection of some antimicrobial agents
administration. (sodium ampicillin or amoxicillin, cefotaxime, genta-
The vein commonly used for injecting drugs intra- micin or amikacin sulfate) in neonatal foals (less than
venously varies with the species of animal. In horses, 7 days of age). This particularly applies in the treat-
cattle, sheep, and goats, the jugular vein is used, ment of septicemia in neonatal foals that are in a state
whereas in dogs and cats, either the cephalic (usually) of septic shock or dehydration or both. The plasma
or the jugular vein is used, and in pigs, an ear vein is concentration profiles for amikacin administered
often used. In anesthetized dogs and cats, the jugular intraosseously and intravenously to neonatal foals
vein is convenient to use for administering supplemen- are similar (Fig. 4).[39]
tal doses of intravenous anesthetics, whereas in dogs,
the sublingual vein may be used in emergency situa-
tions for injecting drug solutions of small volume. In PARENTERAL DOSAGE FORMS
Veterinary–

pharmacokinetic studies of drug bioavailability and

Virtual

disposition, blood samples are collected from the Parenteral dosage forms for use in animals include
jugular vein of all species apart from pigs, in which aqueous, aqueous organic and oily solutions, emul-
the anterior vena cava is generally used. It is very sions, aqueous and oily suspensions for injection, and
unlikely that the vein used for collection of blood sam- modified-release devices for subcutaneous implan-
ples, with the exception of the vein used for injection tation. Parenteral preparations must be sterile and
of the drug, would influence the results obtained in a pyrogen-free; liquid formulations should, if possible,
pharmacokinetic study involving the measurement of be buffered close to physiological pH and preferably
plasma drug concentrations over an extended period. be isotonic with body fluids. The advantages and dis-
Because of practical considerations, intravenous advantages associated with the various types of par-
infusion is used far less widely for drug administration enteral dosage forms have been addressed elsewhere.[40]
to animals than to humans. Intravenous adminis- The formulation of an extravascularly injected
tration, especially constant-rate infusion, has two parenteral preparation primarily determines the
important advantages over other routes. It provides plasma concentration profile of the drug. A drug is
Veterinary Dosage Forms 3955

immediately available for absorption only when admi- should be administered only by the intravenous route.
nistered as an aqueous solution and when neither pre- Formulations containing sparingly soluble drugs in a
cipitation nor tissue irritation occurs at the injection water-miscible solvent, such as propylene glycol, may
site. Gentamicin sulfate (50 mg/ml aqueous solution) cause precipitation of drug at the intramuscular injec-
is rapidly and completely absorbed in that the peak tion site. This makes these preparations unsuitable for
plasma concentration is attained within 30–60 min, intramuscular administration (e.g., diazepam, pheny-
and systemic availability exceeds 90% from intra- toin). Diazepam injection (5 mg/ml solution or emul-
muscular injection sites in horses, dogs, and cats. After sion) may be administered to dogs and cats by
intravenous, intramuscular, and subcutaneous injec- intravenous injection and, after appropriate dilution,
tion of gentamicin (3 mg/kg) in dogs, the peak plasma by intravenous infusion.
concentration (10.7 mg/ml i.m.; 10.2 mg/ml s.c.) and In parenteral preparations containing a poorly
absolute bioavailability (96% i.m.; 94.3% s.c.) of soluble salt of a drug, availability of the drug for
the antibiotic provided by the intramuscular and sub- absorption, which is dependent on dissolution rather
cutaneous routes were similar.[41] Administration of than on the absorption process per se, generally controls
amikacin sulfate (50 mg/ml aqueous solution) by sub- the rate of absorption, the peak plasma concentration,
cutaneous injection at three dose levels (5, 10, and and the length of time over which effective plasma
20 mg/kg) to dogs showed that area under the curve, concentrations are maintained. Long-acting parenteral
which reflects the extent of absorption, was pro- preparations are formulated in a non-aqueous vehicle
portional to the dose administered.[42] After intramus- (such as an oil), or a poorly soluble salt of the drug
cular injection of ceftiofur sodium (as reconstituted is used (usually an aqueous suspension). These pre-
aqueous solution) at dose levels of 1.1 and 2.2 mg of parations provide slow absorption of the drug over
ceftiofur free acid equivalents per kilogram of body an extended period owing to its gradual or staged
weight to sheep, the peak plasma concentrations (Cmax) availability for absorption.[44] Avermectins and milbe-
of ceftiofur and metabolites (measured as desfuroylcef- mycins (macrocyclic lactones) are highly lipophilic sub-
tiofur acetamide by HPLC) were 4.33 and 7.13 mg/ml, stances that determine the extent of their distribution
the apparent half-lives were 6.5 and 7.65 h, respect- and deposition in body fat, whereas the formulation
ively, and the area under the curve (from time zero of parenteral dosage forms (for subcutaneous injection
to the limit of quantification of the assay) was pro- in cattle or sheep, but not horses) influences the plasma
portional to the dose administered.[43] The drug was concentration profile after the administration of a
rapidly (tmax, 0.5–1 h) and completely absorbed from single dose (200 mg/kg). The commercially available
the intramuscular injection site. Ketamine hydrochlor- preparations differ in formulation; ivermectin is a
ide (100 mg/ml aqueous solution) is rapidly absorbed non-aqueous preparation (60% propylene glycol/40%
from the intramuscular injection site in cats, but pain glycerol formal), doramectin is an oil-based prep-
may be evident before onset of the anesthetic effect. aration containing sesame oil/ethyl oleate (90/10),
In horses, dogs and cats, ketamine administration and moxidectin is an aqueous-based solution. The
is preceded by sedative premedication with an a2- pharmacokinetic parameters describing the rates of
adrenoceptor agonist (detomidine, medetomidine, or absorption and elimination of these endectocides after
xylazine). Parenteral solutions of an irritant nature administration by subcutaneous injection (shoulder
(thiopental sodium, ticarcillin sodium-clavulanate region) in 10-month-old Hereford calves are compared
potassium combination, cefuroxime, cefotaxime, cef- in Table 5.[45] Because the systemic availability (F) of
triaxone, flunixin meglumine) and digoxin injection the drugs was not determined in this study, the term

Veterinary–
Virtual
Table 5 Pharmacokinetic parameters describing the rate of absorption and elimination of ivermectin
(IVM), doramectin (DRM), and moxidectin (MXD) after subcutaneous injection (shoulder region) of
single doses (200 mg/kg) of the commercially available preparations to 10-month-old hereford calves
(180–210 kg body weight)
Kinetic parameter IVM DRM MXD
Cmax (ng/ml) 42.8  3.8 37.5  3.9 39.4  3.4
tmax (days) 4.00  3.94a 6.00  1.35 0.32  0.0c
t1/2 (days) 17.2  4.26b 6.25  0.16 14.5  1.20c
a
ClB/F (ml/day  kg) 457  52.5 322  164 938  62.5c
a
Mean kinetic parameters for IVM are significantly different from those obtained for MXD at P < 0.05.
b
Mean kinetic parameters for IVM are significantly different from those obtained for DRM at P < 0.05.
c
Mean kinetic parameters for MXD are significantly different from those obtained for DRM (c) at P < 0.05.
Results are expressed as mean  sem (n ¼ 4).
 3956 Veterinary Dosage Forms

clearance/systemic availability (CLB/F) is used. Com- 5

Ampicillin concentration in µg/ml plasma

parison of three commercially available parenteral pre-
parations (one conventional and two long-acting) of
4
oxytetracycline (20 mg/kg) injected intramuscularly in
the lateral neck of pigs showed statistically significant
differences between the preparations in peak plasma 3
concentration (Cmax), time of peak concentration
(tmax), and mean residence time (MRT), whereas area
2
under the curve (AUC) did not significantly differ
among the preparations (Table 6).[46] The results indi-
cate that a 24-h dosage interval should be used for the 1
conventional preparation, whereas a 48-h interval
would be appropriate for either of the long-acting pre-
parations. A single intramuscular dose (20 mg/kg) of a 0
0 1 2 3 4 5 6 7 8 9 10 11 12 2748
long-acting preparation of oxytetracycline provides Hours after administration
plasma concentrations above 0.5 mg/ml for 48 h in pigs,
ruminant calves, cattle, goats, red deer (Cervus elaphus), Fig. 5 Mean plasma ampicillin concentrations after i.m.
fallow deer (Dama dama), and camels (Camelus drome- injection of 5 different parenteral ampicillin formulations at
darius). The commercially available long-acting prepara- similar dose levels (7.7  1.0 mg/kg) to 5 calves. (From
tions of oxytetracycline should not be administered to Ref.[47].)
Equidae (horses, ponies, and donkeys). When comparing
parenteral preparations of an antimicrobial agent on the is useful. A crossover design incorporating an appro-
basis of potential clinical efficacy, it is generally useful to priate washout period between the phases of the bio-
compare the areas under the inhibitory plasma concen- availability study should be used whenever feasible.
tration–time curves (AUIC ¼ AUC/MIC90) for the The drug concentration in a parenteral suspension
duration of the recommended dosage interval because can influence the plasma concentration profile. When
this term indicates the degree of exposure of susceptible different concentrations of amoxicillin (100 and
micro-organisms to the drug. 200 mg/ml) in aqueous suspensions of amoxicillin tri-
The plasma concentration profiles can vary widely hydrate were administered intramuscularly at the
among different parenteral preparations of the same same site and same dose level (10 mg/kg) to horses,
drug administered by intramuscular injection, even the preparation of lower concentration (10%) provided
when the same injection site is used. Comparison of relatively better absorption and a more consistent
the plasma concentration profiles obtained after intra- plasma concentration profile. Intramuscular injection
muscular injection in the lateral neck of ruminant of amoxicillin trihydrate (15% in a mixed oil base) in
calves of five different parenteral preparations of ampi- the neck (10 cm behind the ear) of pigs produced two
cillin at a similar dose level (7.7  1.0 mg/kg) serves peaks, 1.7 and 0.8 mg/ml at 1.3 and 6.6 h, respectively,
to illustrate the variation that can exist among pre- rather than a single peak in the plasma concentration
parations (Fig. 5).[47] Estimation of the bioavailability profile and an eight-fold longer mean residence time
of each preparation relative to a reference preparation of the antibiotic than a preparation of the same

Table 6 Pharmacokinetic parameters describing the absorption and disposition of three oxytetracycline
Veterinary–

formulations administered intramuscularly (lateral neck) to pigs (n ¼ 8) at a dose of 20 mg/kg body weight
Virtual

Pharmacokinetic term Product A Product B Product C

Cmax (mg/ml) 6.27  1.47 5.77  1.0 4.68  0.61
tmax (h) 3.0 (2.0–4.0) 0.5 (0.083–2.0) 0.5 (0.083–2.0)
AUC (mg h/ml) 79.22  25.02 91.53  20.84 86.64  14.21
MRT (h) 11.48  2.01 25.27  9.22 37.66  15.62
Cp(24 h) (mg/ml) 0.81  0.34 1.01  0.26 0.97  0.29
Cp(48 h) (mg/ml) < LOQ 0.40  0.17 0.50  0.09
LOQ ¼ limit of quantification (0.1 mg/ml).
Product A, Engemycine 10% in polyvinylpyrrolidone.
Product B, Oxyter LA 20% in dimethylacetamide.
Product C, Terramycin LA 20% in pyrrolidone-2 and polyvinylpyrrolidone.
Results are expressed as mean  S.D.
Veterinary Dosage Forms 3957

the ionized moiety, and the extent of binding to plasma

10
proteins and milk macromolecules. It has been shown
8
that only the lipid-soluble, non-ionized moiety of a
mcg/ml plasma

Cat weak organic acid or base that is free (not bound to

6 Piglets (16kg)
proteins) in the plasma can diffuse through cellular
barriers and enter the milk.[51] In normal lactating
Dog (12kg)
4 cows (milk pH range 6.5–6.8), weak organic acids
Calf (43kg)
attain milk ultrafiltrate-to-plasma ultrafiltrate equilib-
2 rium concentration ratios less than 1; oxytetracycline
Horse and rifampin, amphoteric drugs with moderate and
high lipid solubility, attain equilibrium concentration
0 1 2 3 4 5 6 7 8
Hours ratios of 0.75 and approximately 1, respectively; weak
organic bases, apart from aninoglycosides, spectino-
Fig. 6 Effect of species and weight on the bioavailability of mycin, and polymyxin B (drugs with low solubility in
amoxicillin after i.m. injection of amoxicillin trihydrate aque- lipid) attain milk ultrafiltrate-to-plasma ultrafiltrate
ous suspension (100 mg/ml) at the same dose (7 mg/kg) in concentration ratios greater than 1 (Table 7).[52] The
the various species except cats (10–12 mg/kg). high concentration ratios attained by lipophilic organic
bases (macrolides, lincosamides, trimethoprim) are
concentration in oil.[48] The two peaks in the plasma attributed to the ion-trapping effect in acidic milk.
concentration profile could be ascribed to the mixture Enrofloxacin and its active metabolite ciprofloxacin,
of oil vehicles releasing amoxicillin at different rates. formed by N-deethylation (a microsomal-mediated
Age and body weight (45, 55, and 62, kg) of calves oxidative reaction in the liver), would be expected to
was shown to influence the systemic availability of attain concentrations in milk that would be effective
amoxicillin administered intramuscularly as amoxi- against susceptible Gram-negative aerobic bacteria, in
cillin trihydrate (10% aqueous suspension). When the particular Escherichia coli.
same preparation was administered intramuscularly In the presence of mastitis, the pH of milk increases
to different animal species, the trend was for smaller to within the range of 6.9 to 7.2. As a consequence, the
animals (piglets, dogs, cats) to show an early high peak ion-trapping effect on lipophilic organic bases is
concentration followed by a rapid decline, whereas reduced, whereas the concentrations attained by weak
larger animals (calves, horses) show a lower and rela- organic acids are somewhat increased. The higher pH
tively constant plasma concentration of amoxicillin over of milk does not change the concentrations attained
at least an 8-h period (Fig. 6).[49] by amphoteric drugs (fluoroquinolones, tetracyclines,
rifampin); however, antimicrobial activity of these
drugs is often lower in milk than in extracellular fluid
BOVINE MAMMARY GLAND or in vitro determination would predict.
Antimicrobial therapy is generally applied in the
Antimicrobial agents, as with other drugs, cross the treatment of clinical mastitis during lactation and in
blood–milk barrier, which is a somewhat restrictive treating subclinical mastitis at the end of lactation,
functional rather than anatomical barrier, primarily whereas the implementation of preventive measures is
by passive diffusion. Both non-polar lipid-soluble essential for decreasing the incidence of mastitic infec-
compounds and polar substances with sufficient lipid tion in the dairy herd. The common causative
solubility passively diffuse through the predominantly pathogenic microorganisms of clinical mastitis are
lipoidal blood–milk barrier.[50] At a moderate level of
Veterinary–
Streptococcus uberis, coliforms (E. coli, Klebsiella

Virtual
milk production, the ratio of the volume of blood spp.), S. aureus, S. dysgalactiae, and S. agalactiae. It
circulating through the mammary gland to the volume is usual to treat clinical mastitis both systemically using
of milk produced has been estimated to be 670 : 1; a parenteral antimicrobial preparation and locally
this provides ample opportunity for drugs to diffuse with a quick-release intramammary preparation. The
passively from the systemic circulation into milk. The infusion via the teat canal of an intramammary prep-
composition (g/dL) of milk varies among breeds of aration alone would be inadequate for the treatment
cow in protein, from 3.1 (Holstein) to 3.9. (Jersey, of moderate to severe infection because of the
Zebu, and buffalo) and fat, from 3.5 (Holstein) to 5.5 decreased ability of an infused drug to ascend partially
(Jersey, Zebu, and buffalo) content. occluded milk ducts and the requirement, particularly
The milk-to-plasma equilibrium concentration ratio in coliform mastitis, for frequent milkout (stripping)
of total (non-ionized plus ionized) drug is determined of the infected quarter of the mammary gland. In mild
by the degree of ionization of the drug, which is cases of mastitis diagnosed at an early stage of infec-
pKa/pH-dependent, in blood and milk, the charge on tion, the infusion of a quick-release intramammary
 3958 Veterinary Dosage Forms

Table 7 Comparison of calculated and experimentally obtained milk: plasma concentration ratios for antimicrobial
agents under equilibrium conditions
Concentration ratio (milk ultrafiltrate: plasma ultrafiltrate)

Drug Lipid solubility pKa Milk pH Theoretical Experimental

Acids
Penicillin G Low 2.7 6.8 0.25 0.13–0.26
Cloxacillin Low 2.7 6.8 0.25 0.25–0.30
Ampicillin Low 2.7, 7.2 6.8 0.24–0.30
Cephaloridine Low 3.4 6.8 0.25 0.24–0.28
Cephaloglycin Low 4.9 6.8 0.25 0.33
Sulfadimethoxine Moderate 6.0 6.6 0.20 0.23
Sulfadiazine Moderate 6.4 6.6 0.23 0.21
Sulfamethazine Moderate 7.4 6.6 0.58 0.59
Rifampina Moderate/high 7.9 6.8 0.82 0.90–1.28
Bases
Tylosin High 7.1 6.8 2.00 3.5
Lincomycin High 7.6 6.8 2.83 3.1
Spiramycin High 8.2 6.8 3.57 4.6
Erythromycin Very high 8.8 6.8 3.87 8.7
Trimethoprim High 7.3 6.8 2.32 2.9
Aminoglycosides Low 7.8b 6.8 3.13 0.5
Spectinomycin Low 8.8 6.8 3.87 0.6
Polymyxin B Very low 10.0 6.8 3.97 0.3
Amphoteric
Oxytetracycline Moderate — 6.5–6.8 — 0.75
Doxycycline Moderate/high — 6.5–6.8 — 1.53
a
Rifampin is an amphoteric drug substance; theoretical concentration ratio was based on its behavior as an organic acid.
b
pKa value given for aminoglycosides is unconfirmed.

preparation may suffice without concurrent systemic 2. high systemic availability after intramuscular
treatment. Slow-release intramammary preparations injection;
are used at the end of lactation (after the last milking) 3. lipid-soluble and predominantly non-ionized in
to treat subclinical mastitis and to prevent the establish- the blood and have a low degree of binding to
ment of new infections, including summer mastitis plasma proteins;
commonly caused by Actinomyces pyogenes during the 4. long apparent half-life to ensure that concen-
non-lactating (dry) period. Because S. aureus is the prin- trations above (preferably several fold) the MIC
cipal causative microorganism of subclincial mastitis, the are maintained at the site of infection in the
Veterinary–

slow-release intramammary preparation selected for mammary gland throughout the recommended
Virtual

treatment should contain an antimicrobial effective dosage interval (12 h is desirable);

against all strains of the bacterium. 5. minimal adverse effects in cows treated at effec-
tive dosage; and
6. short withdrawal periods (milk and slaughter).

PARENTERAL PREPARATIONS
Parenteral preparations with antimicrobial activity,
An ‘‘ideal’’ antimicrobial agent for systemic therapy of which depends on the causative pathogenic micro-
bovine mastitis should possess the following properties:[53] organism, and pharmacokinetic properties that meet
most of these criteria include procaine penicillin G (aque-
1. low minimum inhibitory concentration (MIC) ous suspension), amoxicillin trihydrate–clavulanate
for the majority of mastitis-causing pathogenic potassium combination (aqueous suspension), and enro-
microorganisms; floxacin (solution). Enrofloxacin is not approved for use
Veterinary Dosage Forms 3959

in cows producing milk for human consumption. Slow-release intramammary preparations may be
To attain effective concentrations in the mammary infused at the end of lactation (after the last milking)
gland, oxytetracycline hydrochloride (conventional and into the teat canal of non-lactating cows to treat
preparation) has to be administered by slow intravenous subclinical mastitis and to prevent the establishment
injection. Even though macrolide antibiotics attain of new infections during the non-lactating period.
high concentrations in milk, they also diffuse passively Either a poorly soluble salt of an antimicrobial agent
into ruminal fluid (pH 5.5–6.5) where the ion-trapping may be used or the formulation of the preparation be
effect applies. This feature of their distribution may such that the rate of antimicrobial release is relatively
be undesirable. Moreover, slow intravenous injection constant, approaching zero-order. The antimicrobial
is the preferred manner of administration because the must remain active (be stable) throughout the extended
available parenteral preparations cause tissue irritation duration in the udder, and the preparation should
at intramuscular injection sites. Because spiramycin not cause tissue irritation. Antimicrobial binding to
binds avidly to tissue components, long withdrawal mammary tissue components is not of particular con-
periods would be associated with the use of this anti- cern because slow-release preparations are not used
microbial agent. in lactating cows. However, the ability to penetrate cell
membranes is important because, in chronic staphylo-
coccal mastitis, the pathogenic bacteria often reside
INTRAMAMMARY PREPARATIONS within epithelial cells, neutrophils, and macrophages.[54]
Examples of slow-release intramammary preparations
Intramammary preparations are formulated to provide include cloxacillin (benzathine salt) with aluminium
either quick release of the antimicrobial agent or slow monostearate (suspension), ampicillin (trihydrate) and
release of the antimicrobial over an extended period. A cloxacillin (benzathine) with aluminium monostearate
requirement of all intramammary preparations is that (suspension) or formulated without aluminium mono-
they be reasonably non-irritating to the parenchyma stearate as an oily suspension, procaine penicillin G
(epithelial tissue) of the udder. Quick-release prepara- (oily paste), dihydrostreptomycin sulfate and procaine
tions are used primarly in lactating cows for the treat- penicillin G (oily paste). Penicillins and especially
ment, often in conjunction with systemic therapy, of aminoglycosides have limited ability to penetrate cell
clinical mastitis. They should have short withdrawal membranes, whereas fluroquinolones, rifampin, and
periods. The vehicle used and viscosity of the formu- macrolides have this capacity.
lation should allow rapid release of the antimicrobial Compound preparations containing one or more
while ensuring that effective concentrations will be antimicrobial agents and a corticosteroid (hydrocorti-
maintained throughout the recommended dosage sone or prednisolone) are available for intramammary
interval. Access of the released antimicrobial to the infusion in lactating cows. The reduction in inflam-
site of infection is determined by its uptake and distri- mation of the mammary gland is desirable; however,
bution in mammary tissue, which are governed by the the immunosuppressant effect and decrease in phago-
chemical nature and physiochemical properties (in cyte function produced by glucocorticoids are undesir-
particular, lipid solubility) of the drug. Binding to able. Amoxicillin trihydrate–clavulanate potassium
milk proteins or components of mammary tissue limits combination with prednisolone is an example of a
distribution and extends the withdrawal periods. compound preparation (oily suspension) that can be
The transfer of an antimicrobial agent from treated administered by intramammary infusion at 12-h inter-
to untreated quarters of the udder takes place via the vals and has short withdrawal periods (slaughter, 7
bloodstream and involves passive diffusion in both days; milk, 2 days). Unlike glucocorticoids, the non-

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directions across the blood–milk barrier. Examples of steroidal anti-inflammatory drugs do not cause

Virtual
intramammary preparations formulated as suspensions immunosuppression. Flunixin meglumine (2.2 mg/kg)
and having a recommended dosage interval of 12 h administered by intravenous injection at 24-h dosage
include cloxacillin sodium, ampicillin sodium–cloxacil- intervals may have a place in the treatment of acute
lin sodium combination, trimethoprim–sulfadiazine E. coli (endotoxin) mastitis. The significance of the
combination, and oxytetracycline hydrochloride (oily antipyretic, anti-inflammatory, and analgesic effects
suspension), whereas erythromycin is formulated as produced by the drug is primarily dependent on the
an intramammary solution. Cefuroxime sodium and stage of the inflammatory process at which treatment
cefoperazone sodium (third-generation cephalosporins) is begun. Early diagnosis of coliform mastitis and
are formulated as an oily paste and oily suspension, prompt initiation of treatment with flunixin greatly
respectively. Quick-release intramammary preparations increase the beneficial effect of the drug. The half-life
have short withdrawal periods, typically slaughter, 7 days of flunixin in cows is 8.1 h and the withdrawal periods
and milk-withholding, 3.5 days, or either may be are short (slaughter, 7 days; milk, 12 h). Flunixin
shorter depending on the preparation. does not interfere with the activity of concurrently
 3960 Veterinary Dosage Forms

administered antimicrobial agents, should antimicro- metabolite) are considered in the development of
bial therapy be applied. There is no evidence to sup- anthelmintic dosage forms and selection of their routes
port the contention that systemic antimicrobial of administration for various animal species.[56]
therapy causes a massive release of endotoxin in cows Because anthelmintics are administered in different
with coliform mastitis. Frequent stripping of the dosage forms to ruminant species (cattle, sheep, and
infected quarter(s) to remove bacteria and cellular deb- goats), pigs, horses, and small animals (dogs and cats),
ris is important, perhaps essential, in coliform mas- a wide variety of preparations are commercially avail-
titis.[55] The slow intravenous injection of oxytocin, able for use in animals.
5–10 U of diluted solution (10 U/ml), facilitates the
completeness of stripping (milkout).

DRUG DISPOSITION

Species variations in the disposition of a drug may be

THERAPEUTIC CONCENTRATIONS attributed to differences in the extent of distribution
and/or the rate of elimination of the drug. Variations
The principal pharmacological effects produced are are usual between ruminant and monogastric species
generally associated with the same range of therapeutic and between herbivorous and non-herbivorous species,
plasma concentrations in domestic animals as in depending on the classification system used for mam-
humans (Table 8). The mechanisms of action of drugs malian species. The rate of elimination of drugs differs
appear to be the same in mammalian species. The cal- widely between homeothermic and poikilothermic
culation of a dosage regimen (dose and dosage inter- species. Little is known regarding drug distribution in
val) for a drug preparation is based on a knowledge poikilothermic species (fish and reptiles).
of the therapeutic concentration range and the phar-
macokinetic parameters that describe bioavailability
and disposition of the drug. Complete systemic avail-
ability can be assumed only when a drug is adminis- EXTENT OF DISTRIBUTION
tered intravenously. Species differences in the dosage
regimen for a drug preparation administered by a Significant differences in the extent of distribution of
particular route can generally, but not always, be drugs, particularly lipid-soluble organic bases, are
attributed to variation among species in pharmaco- usual between ruminant and monogastric species.
kinetic behavior of the drug. Species differences in After parenteral administration, lipophilic bases dif-
susceptibility (dosage requirement) to certain drugs fuse passively from the systemic circulation into rum-
(morphine, xylazine) appear to be attributable to inal fluid (pH 5.5–6.5), where they become ‘‘trapped’’
pharmacodynamic variation (affinity and/or efficacy). by ionization. These drugs are slowly reabsorbed or,
Dosage of anthelmintics, which usually refers to a if they possess functional groups suitable for metab-
single dose, is based on semiquantitative assessment olism by hydrolysis or reduction, they may be partially
of clinical efficacy, although, when appropriate, inactivated by ruminal micro-organisms.
bioavailability and the plasma concentration profile The volume of distribution at steady state (Vd(ss)) is
of the active moiety (parent drug and/or active useful for determining the significance of changes in
the extent of distribution of a drug in the presence of
disease states or by plasma protein binding displace-
ment (drug interaction) and for comparing the extent
Veterinary–

Table 8 Principal pharmacological effect and range of of drug distribution in neonatal and adult animals of
Virtual

therapeutic plasma concentrations of some drugs

the same species. The volume of distribution at steady
Pharmacological Therapeutic state, unlike Vd(area), is at least theoretically inde-
Drug effect concentrations pendent of changes in the rate of drug elimination.[57]
Quinidine Antiarrhythmic 0–6 mg/ml
Procainamide Antiarrhythmic 6–14 mg/ml
Lignocaine Antiarrhythmic 1.5–5.0 mg/ml PLASMA PROTEIN BINDING
Propranolol Antihypertensive 20–80 ng/ml
Digoxin Positive inotropic 0.6–2.4 ng/ml Although the extent of plasma protein binding of a
Phenobarbitone Anticonvulsant 10–25 mg/ml drug varies among domestic animal species, the range
Pethidine Analgesic 0.4–0.7 mg/ml of binding is reasonably narrow in the collective spe-
cies, and the difference among individual species is
Theophylline Bronchodilator 6–16 mg/ml
often not of clinical significance. Species variation in
Veterinary Dosage Forms 3961

the binding of acidic drugs could be attributed to less lipid-soluble, and consequently less widely distri-
differences in the conformation of plasma albumin; buted extravascularly than the parent drug or phase I
the lower concentration of plasma albumin in birds metabolite, and the vast majority are pharmacologi-
than in mammals may generally account for the lower cally inactive. Apart from a few exceptions (e.g., the
binding of acidic drugs in birds. N4-acetyl derivative of most sulfonamides), drug con-
jugates are more water-soluble than the parent drug.
Glucoronide conjugates are especially suitable for
ELIMINATION PROCESSES carrier-mediated active excretion in urine or bile or
both. Species variations in conjugate formation could
In domestic animal species, the liver constitutes 1.25– be attributed to the availability of the conjugating
2.5% of live body weight, and hepatic arterial blood agent, the ability to form the ‘‘activated’’ nucleolide,
flow represents 26–29% of cardiac output. The hepatic or the activity of the transferring enzyme.[60] Fish
portal vein contributes additionally to liver blood flow. may have a low capacity to form the activated nucleo-
The kidneys constitute 0.25–0.6% of live body weight lide uridine–diphosphate–glucuronic acid, which would
and receive 22–24% of cardiac output. In avian species limit glucuronide synthesis. Prodrugs belong to a dif-
and reptiles, the renal portal system contributes to ferent category of drug, in that prodrugs per se are
blood flow to the kidneys. In proportion to organ devoid of activity but are metabolically converted by
weight (mass), the kidneys are more richly perfused phase I, usually hydrolytic reactions to the active
with blood than is the liver. parent drug. Examples of orally administered prodrugs
The liver metabolizes drugs, other foreign chemical include enalapril (enalaprilat), pivampicillin (ampicillin),
compounds (xenobiotics), and certain endogenous netobimin (albendazole), and febantel (fenbendazole).
substances (e.g., steroid hormones, bilirubin) by a var- Metabolic conversion (activation) of these prodrugs
iety of pathways. They include hepatic microsomal- occurs presystemically in that it is brought about by
mediated oxidative reactions, reductive and hydrolytic ruminal micro-organisms or takes place during passage
reactions (phase I), and conjugation (synthetic) reac- through the intestinal mucosa or the liver (first-pass
tions with various endogenous substances (phase metabolism).
II).[58,59] Phase I metabolic reactions occur ubiqui- Ruminal microorganisms are capable of at least
tously and are qualitatively similar in mammals, birds, partially inactivating orally administered drugs (chlor-
and fish, but differ widely and unpredictably among amphenicol, trimethoprim, nitroxynil, digoxin) by
species in the rates at which they take place. Although hydrolytic and reductive reactions. The susceptibility
molecular structure is the primary determinant of the of ruminant animals to toxicity caused by ingestion
phase I reaction that will most likely occur, at least a of plants containing cyanogenetic glycosides (e.g., Prunus
moderate degree of lipid solubility is required for spp., Acacia spp., Eucalyptus cladocalyx) is attributed to
microsomal oxidation. The situation is different with ruminal hydrolysis of the glycosides with liberation of
regard to phase II metabolic reactions because some hydrogen cyanide. Intestinal micro-organisms containing
of these are either defective or absent in certain species, b-glucuronidase can reactivate (by hydrolysis) glucu-
which makes the final pathways of metabolism some- ronide conjugates of drugs excreted in bile. Renal
what predictable (Table 9).[50] Requirements of phase excretion is the principal process for elimination of
II reactions include the presence in a drug molecule drugs that are predominantly ionized in the plasma,
(either parent drug or phase I metabolite) of a polar drugs, and drug metabolites. Extensive (>80%)
functional group that is suitable for undergoing conju- binding to plasma proteins limits the availability of
gation, an endogenous reacting substance (conjugating drugs for glomerular filtration but does not hinder

Veterinary–
agent), and a transferring enzyme. The major transfer- carrier-mediated active tubular secretion. Although

Virtual
ring enzymes are UDP-glucuronyltransferase, sulfo- a drug may enter tubular fluid both by glomerular
transferase, N-acetyltransferase, GSH-S-transferase, filtration and by proximal tubular secretion, its renal
and methyltransferase. Conjugates of drugs are polar, clearance may non-etheless be low when substantial

Table 9 Domestic animal species with defects in certain conjugation reactions

Species Conjugation reaction Major target groups State of synthetic reaction
Cat Glucuronide synthesis –OH, –COOH, –NH2, ¼NH, –SH Present, slow rate
Dog Acetylation Ar–NH2 Absent
Pig Sulfate conjugation Ar–OH, Ar–NH2 Present, low extent
(From Ref.[50].)
 3962 Veterinary Dosage Forms

reabsorption occurs in the distal nephron. Because bladder). Compounds excreted in bile have molecular
reabsorption takes place by passive diffusion, it is weights exceeding 300 and a degree of polarity that
influenced by the concentration of drug and its degree enables them to be transported by a carrier-mediated
of ionization in distal tubular fluid and by the rate process from hepatic parenchymal cells into bile.
of passage of glomerular filtrate through the distal Drugs and drug metabolites (primarily conjugates)
nephron. The degree of ionization is determined excreted in bile enter the duodenum, from which some
by the pKa value of the drug and the urinary pH (depending on their lipid solubility) may be reabsorbed
reaction. The usual urinary reaction in carnivorous by passive diffusion.
species is acidic (pH 5.5–7.0), whereas in herbivorous
species it is alkaline (pH 7.2–8.4). The excretion of
weak organic acids is enhanced under alkaline and RATE OF ELIMINATION
decreased under acidic urinary conditions; the con-
verse applies to weak organic bases. Species variations Half-life is the pharmacokinetic parameter used to
in the rate of elimination of renally excreted drugs measure the overall rate of drug elimination. The
(penicillins, most cephalosporins, aminoglycosides, half-life of most drugs that are primarily eliminated
most diuretics, non-depolarizing neuromuscular block- by hepatic biotransformation varies widely among
ing drugs) are primarily attributable to differences in species (Table 10). The usual trend is that half-life is
the glomerular filtration rate (GFR). Based on inulin shorter in cattle and horses (herbivorous species) than
clearance, mean values of GFR (ml/min  kg) are in dogs and cats (carnivorous species), whereas the
3.96 in dogs; 2.94 in cats; 2.80 in pigs; 2.26, 2.25, and half-life of several drugs is longer in humans than
2.20 in goats, cattle, and sheep, respectively; and 1.65 in domestic animals. There are, however, notable
in horses; the estimated value of GFR in humans is exceptions to this trend, such as the methylxanthines
1.84 ml/min  kg. Urinary pH influences the rate of (caffeine and theophylline) in horses and phenylbuta-
elimination of drugs that are moderately lipid-soluble zone in cattle. The half-life of phenylbutazone in cattle
and undergo elimination both by hepatic metabolism varies from 42–66 h and, unlike in horses, dogs, and
and by renal excretion (amphetamine, trimethoprim, humans, does not appear to be dose-dependent.
phenobarbital, sulfonamides). Because of differences in the rate of hepatic elimination
The rate of hepatocyte secretion of bile in domestic (presumably by biotransformation), the half-life of
animal species is 12–24 ml/kg per day; the lower end of various drugs (sulfamethazine, trimethoprim, ceftiofur/
the range applies to the dog and cat, and the upper end desfuroylceftiofur, closantel, clorsulon) is shorter in
applies to the horse (a species that does not have a gall goats (especially dwarf goats) than in sheep, whereas the

Table 10 Species variations in the half-life of some drugs that are primarily eliminated by hepatic metabolism
Drug Cattle Horse Dog Human
Pentobarbitone 0.8 1.5 4.5 22.3
Thiopentone 3.3 2.5 8.3 11.5
Salicylate 0.8 1.0 8.6 12.0
a a
Phenylbutazone 42–66 4.1–4.7 2.5–6.0 72.0a
Flunixin 6.9 1.9 3.7 —
Morphine — 1.0 0.95 1.9
Veterinary–

Ketamine 0.9 0.7 1.0 2.5

Virtual

Caffeine 3.8 18.2 4.25 4.9

Theophylline 6.9 14.8 5.7 9.0
Norfloxacin 2.4 6.4 3.6 5.0
Enrofloxacin 1.7 5.0 3.4 —
Chloramphenicol 3.6 0.9 4.2 4.6
Metronidazole 2.8 3.9 4.5 8.5
Trimethoprimb 1.25 3.2 4.6 10.6
Sulphadiazineb 2.5 3.6 5.6 9.9
Sulphadimethoxineb 12.5 11.3 13.2 40
a
Half-life is dose-dependent.
b
Half-life may be influenced by urinary pH reaction.
Veterinary Dosage Forms 3963

half-life of some drugs (phenylbutazone, norfloxacin) is agent in birds classified as anseriformes (ducks,
shorter in donkeys than in horses and ponies. geese) and galliformes (chickens, turkeys) but not in
Because phenytoin, valproate, carbamazepine and columbiformes (pigeons, doves).[59]
clonazepam have a much shorter half-life in dogs than Even though exceptions exist, which is inevitable in
in humans (Table 11), the dosage intervals that would view of the variety of metabolic pathways, the half-life
be required for anticonvulsant effectiveness in dogs of the majority of drugs that undergo extensive hepatic
make conventional dosage forms of these drugs biotransformation is shorter in laboratory animal
impractical for therapeutic use.[61] The half-life of species (mice, rats, guinea pigs, rabbits) and Rhesus
phenobarbital and naproxen is 2-fold longer in mongrel monkeys than in domestic animal species and humans.
(mixed-breed) dogs than in Beagles. The long half-life of Consistent with this trend is that the average half-life
naproxen in dogs (74 h in mongrels, 35 h in Beagles) is of antipyrine, a marker substance used to indicate
unusual for a non-steroidal anti-inflammatory drug. hepatic microsomal oxidative activity, is shorter in
Naproxen half-life is 13.9 h in humans, 8.3 h in horses, laboratory animals (0.2–1.4 h) and Rhesus monkeys
4.8 h in minipigs, and 1.9 h in Rhesus monkeys. (1.2 h) than in domestic animals (1.75–3.25 h) and
The half-life of salicylate, which is primarily elimi- especially humans (10.3–12.7 h). It has been hypothe-
nated by glucuronide conjugation, is 25–35 h and sized[63] that the lesser capacity of humans to metabo-
dose-dependent in cats, compared with 10–15 h in lize drugs by hepatic microsomal oxidation may be
humans, 8.6 h in dogs, 1 h in horses, and 0.8 h in cattle. correlated with their enhanced longevity, expressed as
A relative deficiency of hepatic microsomal glucuro- maximum lifespan potential. Based on an equation
nyltransferase activity appears to be characteristic of using brain weight and body weight, maximum life-
Felidae, because it applies not only to the domestic span potential is estimated to be 93 y in humans, 39 y
cat (Felis catus) but also to the lion (Panthera leo), in horses, 20 y in dogs, 14 y in cats and, for compara-
African civet (Viverra civetta), and forest genet tive purposes, 77 y in African elephants.
(Genetta pardina).[62] Even though dogs and foxes As with the situation in mammals, wide variations
(Canidae) are unable to acetylate aromatic (ArNH2) exist among avian species in the half-life of drugs
and hyrazine (RNHNH2 or ArNHNH2) amino groups, that are primarily eliminated by hepatic biotrans-
the absence of this metabolic pathway does not appear formation.[64] The half-life of antimicrobial agents is
to delay the elimination of drugs with these functional prolonged in poikilothermic species (fish and reptiles),
groups (sulfonamides, procainamide). Either a larger which is consistent with their much lower metabolic
fraction of the systemically available dose is excreted turnover rate,[65] and is influenced by ambient (in the
as parent drug in the urine or an alternative metabolic case of fish, water) temperature (Table 12). The rate
pathway compensates for the absence of acetylation of of drug elimination increases (i.e., half-life decreases)
these amino groups. Amino acid conjugation in with increase in ambient temperature and varies
domestic animal species often involves glycine as the among fish species.
endogenous reacting substance. Glycine conjugation Species variations in the half-life of drugs that are
takes place in both the liver and the kidneys of dom- eliminated by renal excretion is less pronounced than
estic animals other than the dog, in which it occurs for lipid-soluble drugs that undergo extensive hepatic
only in the kidneys. The transferring enzyme, which biotransformation. The half-life of gentamicin, which
is located in mitochondria, is acyl-CoA glycinetransfer- is eliminated solely by glomerular filtration, is 0.5–1 h
ase. Glycine is replaced by ornithine as the conjugating in laboratory animals, 1.25–2.5 h in domestic animals,

Veterinary–
Virtual
Table 11 Comparison of the average half-life (h) after intravenous administration (apart from
carbamazepine) of a single dose of anticonvulsant drugs in dogs and humans
Therapeutic range of plasma
Drug Dog Human concentrations (lg/ml)
Phenobarbitone 64 96 10–25
Phenytoin 3.5–4.5a 15–24a 10–20
Sodium valproate 2 14 40–100
Carbamazepine (PO) 1.5 15 4–10
a a
Clonazepam 1.5–2.5 24–36 0.01–0.08
Diazepam 7.6b 32.9b >0.15
a
Half-life is dose-dependent.
b
Parent drug and active metabolites.
 3964 Veterinary Dosage Forms

Table 12 Half-life of some antimicrobial agents in various species of fish

Acclimatization
Antimicrobial agent Fish species temperature ( C) Half-life (h)
Trimethoprim Carp 10 40.7
(Cyprinus carpio L.) 24 20.0
Sulphadiazine Carp 10 47.0
(Cyprinus carpio L.) 24 33.0
Sulphadimidine Carp 10 50.3
(Cyprinus carpio L.) 20 25.6
Rainbow trout 10 20.6
(Salmo gairdneri) 20 14.7
Ciprofloxacin Rainbow trout 12 11.2
(Salmo gairdneri)
Carp 20 14.5
(Cyprinus carpio)
African catfish 25 14.2
(Clarias gariepinus)
Florfenicol Atlantic salmon 10.8  1.5 12.2
(Salmo salar) (Sea water)
Oxytetracycline Rainbow trout 12 89.5
(Salmo gairdneri)
African catfish 25 80.3
(Clarias gariepinus)
Gentamicin Channel catfish 22 12.0
(Ictalurus punctatus)

2.75 h in humans, 1.25–3.4 h in various avian species, of body weight in humans and dogs, respectively. It
and 12 h in channel catfish (Ictalurus punctatus) accli- can be concluded that another organ (the lungs) or
mitazed at 22 C. In mammalian species, the half-life of extrahepatic tissue contributes to the elimination
gentamicin reflects the relative (not actual) rate of (metabolism) of propofol. The blood propofol concen-
glomerular filtration and is unrelated to urinary pH tration at which dogs return to the sternal position and
reaction. The volume of distribution of gentamicin is human beings regain consciousness appears to be the
similar (250–300 ml/kg) in the various species. same (1 mg/ml).
The half-life of drugs that undergo a high degree of When applied to conventional (immediate-release)
enterohepatic circulation may vary widely among spe- aminophylline tablets, the dosing rates that would
cies and may be relatively more prolonged in fish. The provide an average steady-state plasma theophylline
half-life of oxytetracycline is 80.3 h in African catfish concentration of 10 ug/ml and produce a sustained
(Clarias gariepinus) acclimitazed at 25 C and 89.5 h bronchodilator effect are 10 mg/kg administered at
in rainbow trout (Salmo gairdneri) at 12 C[66] 8-h dosage intervals to dogs and 5 mg/kg administered
compared with half-life of 9 h in humans, 3.4–9.6 h in at 12-h intervals to horses or cats. The systemic clear-
Veterinary–

domestic animals, and 1.3 h in rabbits. A combination ance of theophylline is 2.5 times higher in dogs
Virtual

of variables, which include the extent of distribution (100 ml/h  kg) than in horses and cats (40 ml/
and the degree of enterohepatic circulation, contributes h  kg), whereas systemic availability of the drug
to species variation in the half-life of digoxin, which from the oral dosage form is similar (90–100%).
ranges from 7.8 h in cattle to 35 h in cats.

INTERSPECIES SCALING
CLEARANCE
The basic assumption in interspecies scaling is that
The systemic clearance of propofol, based on measure- physiological variables and biochemical processes are
ment of blood concentrations, in humans (31 ml/ related to the body weight of mammalian species.
min  kg) and dogs (59 ml/min  kg) exceeds liver The use of clearance, a physiologically based param-
blood flow, which is 24 and 42 ml/min per kilogram eter, may be more appropriate than half-life (a hybrid
Veterinary Dosage Forms 3965

parameter) for interspecies allometric scaling of drug Because the clearance of highly extracted drugs that
elimination. A double logarithmic plot of the pharma- undergo extensive phase I hepatic metabolism is
cokinetic parameter of interest vs. body weight of primarily determined by a single physiological variable
the animal species is used to verify the linearity of (liver blood flow), interspecies allometric scaling
the allometric relationship. should be feasible for drugs in this category. The index
The systemic clearance of structurally unrelated of drug elimination (half-life, systemic clearance, organ
drugs that undergo elimination by various processes clearance of unbound drug) to use depends on the level
shows a high degree of correlation with the body of refinement required.
weight of several animal species (Table 13). Inulin With regard to organ (liver, kidney, heart) weights,
(marker substance) and gentamicin are eliminated by physiological variables (liver blood flow, renal func-
glomerular filtration and ampicillin by glomerular tion, cardiac output, basal oxygen consumption), and
filtration and proximal tubular secretion, whereas the pharmacokinetic parameters systemic clearance
oxytetracycline undergoes enterohepatic circulation and volume of distribution, the numerical value of
and consequently is slowly excreted by glomerular fil- the allometric exponent is generally in the range of
tration. Enrofloxacin, theophylline, and antipyrine 0.67 to 1. For physiological periods(heartbeat time,
are poorly extracted by the liver, which implies that duration of respiratory cycle), turnover times (serum
their clearance may be influenced both by the unbound albumin, total body water, blood circulation), and
fraction in blood and by the metabolic capacity of the drug half-life, the allometric exponent is often close
liver and are eliminated by phase I hepatic metabolism. to 0.25, which represents that for energy expenditure
Enrofloxacin and theophylline are eliminated primarily in mammalian species[69] and the turnover time of
by microsomal-mediated oxidative reactions, whereas endogenous processes.[63]
antipyrine (marker substance) is eliminated entirely The reliability of predictions based on interspecies
by microsomal oxidation. Interspecies scaling of allometric scaling of drug disposition depends on the
antipyrine clearance identifies the human as a non- use of a sensitive and precise analytical method for
conforming species, which is attributed to the substan- quantification of the drug (and active metabolite) in
tially lower microsomal oxidative capacity of humans blood or plasma, a knowledge of the principal elimin-
relative to the 10 other species studied. Because of ation process for the drug, the use of at least four
species variations in binding to plasma proteins, the animal species representing a wide range of body
use of hepatic intrinsic clearance of unbound drug, weight (based on log body weight ratio), and the
rather than systemic clearance, would represent a identification, for possible exclusion, of any non-
refinement of the allometric scaling technique. This conforming species. Because of the uniqueness of each
refinement has been applied to interspecies scaling of animal species, application of the technique for pre-
antipyrine[67] and theophylline.[68] Additional correc- dictive purposes should be limited to the preclinical
tion could be made for maximum lifespan potential.[63] stage of drug development.

Table 13 Allometric relationship between the clearance (ml/min) of some drugs and body weight (kg) of various
mammalian species
Allometric

Drug Elimination process No. of species Coefficient Exponent Correlation coefficient

a

Veterinary–
Inulin E (r) 8 4.13 0.86 0.989

Virtual
a
Gentamicin E (r) 8 2.60 0.86 0.970
Ampicillin E (r) 8b 5.47 0.94 0.959
c
Oxytetracycline E (r) 8 7.96 0.73 0.978
Enrofloxacin M (h) 8d 12.53 0.93 0.984
Theophylline M (h) 9e 1.98 0.83 0.978
Antipyrine M (h) 10f 8.16 0.85 0.989
a
Cat, dog, goat, sheep, pig, human, cattle, horse.
b
Rabbit, dog, sheep, pig, human, donkey, cattle, horse.
c
Rabbit, dog, goat, sheep, pig, donkey, cattle, horse.
d
Rabbit, cat, dog, sheep, pig, ilama, horse, cow.
e
Rat, guinea pig, rabbit, cat, dog, pig, human, cattle, horse.
f
Mouse, rat, guinea pig, rabbit, monkey, dog, goat, sheep, pig, cattle (human is a non-conforming species).
For each drug, the level of significance is P < 0.001.
 3966 Veterinary Dosage Forms

IMPLICATIONS OF STEREOISOMERISM (Table 14), whereas values of the pharmacokinetic

parameters in calves did not differ between the enan-
Stereoisomerism has implications in the formulating of tiomers.[71–73] The S(þ)- to R(
) ratio of area under
dosage forms and because a chiral environment exists the curve was 1.35 : 1 in horses, 0.54 : 1 in sheep, and
within the body, in determining both the degree of 1.05 : 1 in calves. The predominant enantiomer in
activity and the disposition of racemates. The enantio- plasma was S(þ) in horses, R(
) in sheep, and both
selective behavior of drugs used in domestic animals enantiomers were present in equal concentrations in
was comprehensively reviewed by Landoni et al.[70] calves. After the administration of each enantiomer
Because stereoselective processes are species-related, separately to these species, the extent of chiral inver-
the enantiomeric ratios of plasma concentrations sion from the R(
)- to S(þ)-enantiomer was estimated
at various times and areas under the plasma concen- to be 49% in horses, 5.9% in sheep, and 31% in
tration–time curves may differ among animal species calves.[72,74,75] Unidirectional chiral inversion was
after the administration of a drug racemate. Chiral estimated to be 49% in Cynomolgus monkeys
inversion, which occurs to a variable extent in different (Macaca),[76] 9% in humans,[77] and varied from 27 to
species, can be equivocally established only by admin- 66% in laboratory animal species.[78] The oral bioavail-
istering individual enantiomers to the animal species of ability of the S(þ)-enantiomer of ketoprofen in Beagle
interest and measuring, using a sensitive stereospecific dogs is not affected by the proportion of the R(
)-
analytical method, the enantiomer administered and enantiomer in the oral dosage form, even though con-
the optical antipode in biological fluids and tissues. siderable (73%) metabolic inversion from the R(
) to
The pharmacokinetic parameters based on plasma the S(þ)-enantiomer occurs in dogs.[79]
concentration–time data for each of the enantiomers It is usual in humans for the S(þ)-enantiomer of
can be statistically compared. 2-arylpropionic acids to predominate in plasma and for
The 2-arylpropionic acid (‘‘profen’’) non-steroidal the S(þ)- to R(
)-enantiomeric ratio of plasma con-
anti-inflammatory drugs, each of which contains a centrations to increase with time after administration
single chiral center, are formulated as racemic of the racemate, which is often attributed to metabolic
(50 : 50) mixtures of the S(þ)- and R(
)-enantiomers, inversion of the chiral center of the R(
)-enantiomers
with the exception of naproxen, which is formulated to their S(þ)-antipodes.[80] In humans, the S(þ)-
as the S(þ)-enantiomer. Based on inhibition of cyclo- enantiomer is generally eliminated more slowly than
oxygenase activity, the S(þ)-enantiomer is the eutomer is the R(
)-enantiomer. The extent of chiral inversion
(more potent enantiomer). These drugs differ markedly of fenoprofen, which has been attributed to the differ-
in both pharmacodynamic activity and pharmacoki- ential rate of formation of the CoA-thioester by
netic behavior and, in addition, enantiomer pharmaco- hepatic microsomes,[81,82] varies widely among species.
kinetics of each drug varies among animal species. It has been estimated to be 90% in dogs,[83] 80% in
After intravenous administration of racemic keto- sheep,[81] 73% in rabbits,[84] 60% in humans,[85] 42%
profen to horses, sheep, and 20-week-old calves and in rats,[86] and 38% in horses.[83]
measurement of individual enantiomers in plasma, Carprofen, a weak inhibitor of cyclo-oxygenase but
significant differences between the enantiomers were which produces a significant antioedematous effect
found in systemic clearance in horses and in both sys- in dogs[87] and horses[88] and has potent antiplatelet-
temic clearance and volume of distribution in sheep aggregating properties, does not appear to undergo

Table 14 Pharmacokinetic parameters describing disposition of S(þ)- and R(
)-enantiomers after intravenous administration
Veterinary–

of racemic ketoprofen (KTP) to horses (n ¼ 6) and sheep (n ¼ 6)

Virtual

Horses Sheep

Pharmacokinetic parameter S(þ)-KTP R(
)-KPT S(þ)-KTP R(
)-KTP

t1/2(a) (h) 0.13  0.03 0.10  0.02 0.14  0.01 0.13  0.03
t1/2(b) (h) 1.51  0.45 1.09  0.19 0.86  0.08 0.87  0.10
Vd (ml/kg) 491  206 472  146 256  21 168  15a
CLB (ml/h  kg) 202  22 277  35a 351  50 196  32a
MRT (h) 2.23  0.15 2.63  0.33 0.79  0.11 0.95  0.13
a
AUC (mg  h/ml) 5.67  0.47 4.19  0.37 4.74  0.71 8.73  1.22a
a
P < 0.05.
(From Refs.[71,72].)
Values are expressed as mean  S.E.M.
Veterinary Dosage Forms 3967

chiral inversion in either direction in horses, calves, may influence oral bioavailability of the enantiomers
dogs, cats, and humans. After intravenous or oral of chiral drugs that are highly extracted by the liver
administration of racemic carprofen, which contains and administered as racemic mixtures. When both
a 50 : 50 mixture of the enantiomers, to horses (i.v.), enantiomers of a drug with a single chiral center show
8- to 10-week-old calves (i.v.), cats (i.v.), and dogs distinct and desirable effects (e.g., most opioids, dobu-
(p.o.), the R(
)-enantiomer predominated in the tamine, bupivacaine), even though they differ in phar-
plasma and the R(
)- to S(þ)-enantiomeric ratio of macodynamic activity or when their action and the
plasma concentrations increased with time after effects produced are not stereoselective, the formulat-
administration of the racemate. The increasing ing of racemic mixtures may be entirely justifiable.[98]
R(
) : S(þ) enantiomeric ratio of plasma concentra- Nonetheless, because of species variation, pharmacoki-
tions with time can be attributed to stereoselective netic profiles of the individual enantiomers should be
hepatic metabolism, although stereoselective binding determined using stereospecific analytical methods[99–102]
to plasma albumin could contribute. A contrasting to calculate optimum dosage for the various animal
situation to that which occurs in domestic animals species. Use of the more active enantiomer (eutomer)
was found in rats and humans, in that the S(þ)-enan- in formulating dosage forms should be considered when
tiomer predominated in plasma and the R(
) : S(þ) the enantiomers differ widely in pharmacodynamic
enantiomeric ratio of plasma concentrations decreased activity [e.g., S(
)-propranolol, the S(þ)-enantiomer
(although only slightly in humans) with time after of the 2-arylpropionic acid non-steroidal anti-inflamma-
administration of the racemate.[89,90] The R(
)- to- tory drugs, D-propoxyphene] or toxic potential (levami-
S(þ) ratio of area under the plasma concentration–time sole is the L-isomer of racemic tetramisole, which can
curve (AUC) was 4.5 : 1 in horses,[91] 2.0 : 1 in cats,[92] produce many side effects). To selectively produce a
1.8 : 1 in dogs,[87] and 1.4 : 1 in calves,[93] after administra- certain effect, a distomer (less potent enantiomer) could
tion of the racemate. Area under the curve enantiomeric be formulated as a particular dosage form, e.g., R(þ)-
ratios in rats and humans were not determined. timolol as eyedrops to reduce intraocular pressure
Ketamine, which is present in the commercially (glaucoma), R(þ)-verapamil as a parenteral or oral
available preparation as a 50 : 50 mixture of the S(þ)- dosage form for the treatment of angina.[103] The use
and R(
)-enantiomers, is metabolized by hepatic of an enantiomer, a single chemical entity, would
microsomal N-demethylation to the corresponding increase selectivity of action, reduce total exposure to
norketamine (metabolite I) enantiomers. Based on the racemate, and simplify dose–response relationships.
the reported eudismic ratio of S(þ) : R(
) for ketamine When an enantiomer is used in formulating dosage
enantiomers of 2.9 : 1 and the observed duration of forms, it must be optically pure. Bioequivalence assess-
unconsciousness in dogs and the plasma concentra- ment of a generic drug preparation requires that the
tions in humans at time of emergence from anesthesia generic preparation contain the racemic drug or the
which are 0.5 [S(þ)] and 1.7 [R(
)] mg/ml, it can be enantiomer corresponding to whichever is present in
concluded that the S(þ)-enantiomer is three times the innovator (reference) dosage form.
more active than is the R(
)-enantiomer.[94,95] After
intravenous injection of racemic ketamine, the S(þ)-
enantiomer of norketamine predominated in the plasma PERCUTANEOUS ABSORPTION
of horses[96] and dogs.[97] This could be attributed to
enantioselective N-demethylation. Systemic clearances The skin accounts for approximately 10% of live body
of racemic ketamine are 15, 28, and 29 ml/min  kg in weight in cattle, goats, and dogs; 7.5% in horses; and
humans, horses, and dogs, respectively. Because the dis- 3.7% in humans. Although skin receives approximately

Veterinary–
position of the individual enantiomers administered sep- 6% of cardiac output, cutaneous blood flow rate to

Virtual
arately has not been studied, comment cannot be made various regions differs among species.[104] In humans
regarding the extent of chiral inversion. and pigs, the cutaneous circulation supplies blood
Whether a racemate or an enantiomer of a chiral (in musculocutaneous arteries) to both the skin and
drug should be used in formulating dosage forms the underlying musculature, whereas in dogs and
depends on the relative pharmacodynamic activity cats (loose-skin species), blood is supplied directly to
and the potential toxicity (or side effects) of the indi- the skin.
vidual enantiomers, their pharmacokinetic profiles,
and, importantly, the proportions formed over time
in the target animal species. Binding to plasma and ABSORPTION PROCESS
tissue proteins, hepatic microsomal oxidative reactions,
and probably glucuronide conjugation and carrier- The absorption process for a topically applied drug
mediated excretion processes are stereoselective and essentially involves the following stages: dissolution
vary among animal species. First-pass metabolism of the drug in and release from the vehicle, drug
 3968 Veterinary Dosage Forms

penetration (by diffusion) through the stratum, and exocrine secretions may enhance dissolution and thus
permeation through the ‘‘living’’ layers of the epider- facilitate percutaneous absorption of topically applied
mis to the underlying dermis where absorption into compounds in cattle and sheep. Seasonal changes in
the systemic circulation takes place. Although the the composition of secretions may cause variations in
initial stage is formulation-dependent in that it relates absorption of moderately lipid-soluble drugs (e.g.,
to the form of the drug and the nature of the vehicle, levamisole applied by ‘‘pour on’’) at different times
the translocation stages are primarily governed by the of the year.[109] Because of the larger number of skin
molecular structure and physiocochemical properties appendages per unit of surface area, the appendageal
of the drug. Penetration of the stratum corneum is path will likely contribute more to percutaneous
generally the rate-limiting step in the absorption pro- absorption of hydrophilic substances in cattle and
cess.[105] Only lipid-soluble drugs can diffuse through sheep than in other species. Horses and humans have
the dead, compacted, keratinized cells (corneocytes) highly effective sweat glands, whereas cattle, sheep,
of the stratum corneum. However, passive diffusion pigs, dogs, and cats are unable to sweat profusely.
through the epidermis, including the stratum corneum, Changes in ambient temperature appear to affect
can take place by one, or more, of the following animal skin temperature to a higher degree than
routes: transcellular through the corneocytes, inter- human skin temperature, which suggests that skin
cellular through the lipid matrix (a tortuous path), plays a greater role in thermoregulation in humans
or along the sweat gland ducts and hair follicles than in animals.
(appendageal path). Even though lipophilic sub- The state of hydration of the stratum corneum,
stances may penetrate the stratum corneum by trans- which is normally maintained at 10–15%, affects the
cellular diffusion, some degree of water solubility is rate of penetration of chemical substances. By increas-
required for passage through the ‘‘living’’ layers of ing the state of hydration to 50%, the rate of per-
the epidermis. Polar drugs have a low capacity to meation of some chemical substances through the
penetrate the stratum corneum but may gain access epidermis can be increased up to 10-fold.[110] Occlusion
to the ‘‘living’’ epidermal layers by shunt diffusion has been shown to enhance the pharmacological effect
along the appendageal path. Additional factors that of topically applied hydrocortisone and fluocinolone
influence percutaneous absorption include the nature acetonide;[111] however, percutaneous absorption of
of the vehicle, the state of hydration of the stratum drugs is not necessarily increased. The degree of
corneum, drug persistence in the stratum corneum occlusion-induced absorption enhancement appears
or other strata of the epidermis, biotransformation to increase with increasing lipophilicity of drug sub-
in the epidermis, and species differences in histologi- stances.
cal structure of skin. In aquatic mammals, the stra- Formulations containing an absorption-promoting
tum corneum is very thick, and the corneocytes are substance, such as propylene glycol or sodium lauryl
solidly apposed, whereas the epidermis is devoid of sulfate, may increase the permeability of the stratum
a stratum granulosum regardless of whether the skin corneum to water-soluble drugs. Propylene glycol is
is glabrous (as in whales) or hairy (as in seals).[106] a commonly used vehicle in topical corticosteroid
The average thickness of the stratum corneum in preparations for veterinary use. Various aprotic sol-
laboratory and domestic animals and humans is in the vents, which include dimethylacetamide, dimethyl-
range of 10 to 35 mm. The thickness of this epidermal formamide, dimethylsulfoxide, tetrahydrofurfuryl
layer might not influence the penetration of chemical alcohol, and 2-pyrrolidone, serve as penetration enhan-
substances, whereas the density of appendages per unit cers of polar drugs.[112] Dimethylsulfoxide (DMSO) is
surface area, which differs among animal species, does used in formulating some topical veterinary prepara-
Veterinary–

influence passage through the epidermis. Human skin tions. The penetration-enhancing property of DMSO
Virtual

contains an average of 40–70 hair follicles and 200– is markedly concentration-dependent. At concentra-
250 sweat glands per square centimeter, whereas cattle tions below 50% DMSO is water, the penetration rate
skin contains approximately 2000 hair follicles, with of many drugs differs little than from aqueous solu-
associated sweat and sebaceous glands, per square cen- tions. The penetration rate of levamisole through skin
timeter.[107] The mean follicle density in 10 British of cattle and sheep was somewhat slower from a for-
breeds of sheep varies from 1000 to 2000 per square mulation containing DMSO (concentration not speci-
centimeter of skin, with a secondary-to-primary follicle fied) than from an aqueous solution of the drug.[107]
ratio of 2.4 : 1–5.9 : 1.[108] The ratio of secondary to pri- Plasma and gastrointestinal fluid concentrations of
mary follicles appears to be higher in Merino sheep levamisole were lower after pour-on application to
skin in wool-growing regions. The sebaceous glands cattle than after oral administration or subcutaenous
of cattle and sheep exude large quantities of lipoid injection of the drug.[109] Parathion penetrated the
material (lanolin in the case of sheep) that serve to skin of pigs more rapidly when formulated in DMSO
protect their skin. The emulsifying properties of than in other vehicles (glycerol–formal/isopropranol
Veterinary Dosage Forms 3969

Table 15 Pharmacokinetic parameters for parathion (50 mg/kg) applied topically in various vehicles to pigs
Vehicles
a
Pharmacokinetic parameter GFI DMSO Octanol Macrogol 400
AUC (mg  h/L) 1460–1795 1630–3050 2010–3310 595–600
MRT (h) 57–106 9.7–14.5 22–31 54–60
MAT (h) 55–104 7.5–12.5 20–29 52–58
Bioavailability (%) 16–20 19–28 15–29 3.9–54
Bioavailability (absolute) was based on AUCtopical/AUCIV, with correction for dose.
a
GFI ¼ glycerol–formal/isopropanol mixture.
(From Ref.[113].)

mixture, octanol, macrogol 400) (Table 15).[113] The pig skin occurred more slowly than through skin of
formation of a stable 2 : 1 water hydrate at a concen- the other species. Even though pig skin and human
tration of 67% v/v DMSO may explain its dehydrating skin are similar in many respects,[116] percutaneous
and penetration-enhancing effects when present at high absorption of a variety of compounds in the pig was
concentration.[114] These effects are accompanied by, found to range from zero to four times that in humans
or perhaps attributable to, epidermal tissue damage. in vivo.[117] Advantage can be taken of the often similar
Mineral oil may be used in formulating long-acting, permeability characteristics of pig and human skins
water-based topical preparations of synthetic pyre- with avoidance of the systemic/fat distribution differ-
throids (permethrin, cypermethrin) for application to ence in vivo by using the isolated perfused porcine skin
ruminant animals. This type of preparation would flap in vitro model.[118,119] The diffusion of chemical
not be washed off by rain and could provide protection substances through skin and metabolism within the
against flies for an extended period. Water-insoluble skin can be determined by assay of the perfusate.
substances can be formulated as emulsifiable concen- The sum of the amount of compound that diffused into
trates. The emulsifiable concentrate contains one or the perfusate and the residual amount in the skin prep-
more surfactants and produces an emulsion or micellar aration at the end of the exposure period to the drug
solution with the water-insoluble drug in the non- provide an estimation of percutaneous absorption.
aqueous phase when mixed with water.[107] Regional variations in percutaneous absorption
contribute to differences in the systemic availability
of a drug depending on the site of topical application.
The Rhesus monkey (Macaca mulatta) could probably
SPECIES VARIATIONS serve as an animal model for human skin regional
variation.[120]
The barrier properties of skin vary with the species of The absolute bioavailability of a topically applied
animal and within a species may differ between regions drug can be determined only by measurement of
of the body. Based on limited data obtained from in plasma concentrations and comparing total areas
vitro studies of skin permeability, it could be specu-
lated that species can, in general, be ranked in the fol-
lowing order: rabbits > rats > guinea pigs > cats > Table 16 Maximal penetration of radiolabeled

Veterinary–
dogs > pigs and Rhesus monkeys > humans (least per- organophosphorus compound through excised

Virtual
meable skin). Because of a lack of data, horses, cattle, skin from dorsal thorax of various species
sheep, and goats are not included in the comparison
Species Rate (lg/cm2/min)
of skin permeability. The emulsifying property and
occlusive effect of sebum and the high density of Pig 0.3
appendages per unit of surface area would be expected Dog 2.7
to facilitate percutaneous absorption of substances in Monkey 4.2
ruminant species. Goat 4.4
The maximal rate of penetration of an organophos- Cat 4.4
phorus compound through skin sections excised from
Guinea pig 6.0
the dorsal thorax of various species generally supports
the skin permeability ranking of species vide supra Rabbit 9.3
(Table 16).[115] The compound rapidly penetrated the Rat 9.3
skin of rabbits and rats, whereas penetration through (From Ref.[115].)
 3970 Veterinary Dosage Forms

under the curves or, less reliably, the amounts excreted dehydrogenase (a non-microsomal enzyme); however,
in urine over a period of at least six half-lives after top- whether the enzyme is present in skin does not appear
ical application and intravenous injection of the drug. to be known.
An appropriate washout period must be allowed to
elapse between the phases of a crossover study, which
is the experimental design that should be used when- TOPICAL PREPARATIONS
ever feasible. Because of species variations in ultra-
structure of skin, cutaneous blood supply, density of There is a wide variety of veterinary drug preparations
appendages per unit of surface area, and activity of available for topical application to the skin. Although
biotransformation pathways, the percutaneous absorp- most of these preparations are intended to produce
tion (rate and extent) of a drug is best determined by local effects, some are formulated to distribute via
performing the study in the species of interest. the systemic circulation to skin covering all regions
of the body or to produce systemic effects. Because
of the prevalence of ectoparasites, several ectoparasiti-
CUTANEOUS BIOTRANSFORMATION cidal preparations are available for topical application
by various methods depending on the animal species
Epidermal cytochrome P450 and hydrolytic enzymes in (Table 17). Liquid concentrates, which must be appro-
the lipid matrix of the stratum corneum as well as in priately diluted before use, and prepared solutions are
the stratum granulosum may be involved in conversion convenient to apply and, depending on susceptibility of
reactions of topically applied steroids.[121] In pigs, epi- the ectoparasite and persistence of the drug in skin,
dermal cytochrome P450 has been shown to convert may provide effective treatment and protection against
parathion to paraoxon by oxidative desulfuration.[122] reinfestation for an extended period. The usual method
The oxygen analog formed is rapidly hydrolyzed to of application of prepared solutions is by pour on to
inactive metabolites. The significant difference between cattle, sheep, pigs, and horses, whereas ‘‘spot on’’ is
mammals and insects in the rate at which the hydro- preferred for dogs and cats. The active ingredient in
lytic reaction takes place accounts for the selective tox- preparations for pour-on application must be suffi-
icity of thiophosphate insecticides. The extent to which ciently lipid-soluble for percutaneous absorption to
the ultrastructural difference in the epidermis of aqua- occur, and the preparation should provide residual
tic and terrestrial mammals affects the activity of drug- activity in the stratum corneum and stratum germina-
metabolizing enzymes in skin is not known. Fish appear tivum and not be removed from the skin by environ-
to be unable to detoxify thiophosphate insecticides. mental conditions (such as rain) or by rubbing.
Benzoyl peroxide, the active ingredient in some Diluted liquid concentrates are applied by spray to
shampoos for dogs, is almost completely metabolized cattle, horses, pigs, and poultry and by either dip
to benzoic acid in the epidermis. Benzoic acid under- or spray to sheep. Because of the large quantity of
goes conjugation with glycine, primarily in the liver, drug required for dipping sheep and the difficulty of
and is excreted in urine as hippuric acid. Methylation safe disposal of dip, application of diluted liquid
of norepinephrine to epinephrine, an N-transferase- (dip) concentrate as a spray, but at a higher concen-
mediated conjugation reaction, in human and animal tration than that used in the dip bath, is becoming a
skin preparations has been reported.[123] popular method of ectoparasiticide application by
Biotransformation of propranolol, which involves sheep farmers. The relative effectiveness of spraying
microsomal-mediated oxidative reactions and glucuro- versus dipping sheep for the treatment and prevention
nide conjugation, is stereoselective in both the liver and of ectoparasite infestation is open to question. When
Veterinary–

the skin. Based on in vitro studies, using intact human choosing between an organophosphorus compound
Virtual

skin and microsomal preparations, of percutaneous (dimpylate, propetamphos) and a synthetic pyrethroid
absorption and metabolism of racemic propranolol, it (flumethrin, cypermethrin) for dipping sheep, a con-
was concluded that the S(
)-enantiomer (eutomer) is sideration (in addition to residual protection) that
metabolized more efficiently by skin than is the could be of practical relevance is that there is no with-
R(þ)-enantiomer.[124] The converse applies to hepato- drawal period associated with the use of synthetic pyr-
cytes, which metabolize the R(þ)-enantiomer more ethroids. The low toxicity of pyrethroids in mammals
efficiently.[125] The cytochrome P450 mono-oxygenase is primarily attributed to the rapid biotransformation
enzymes in skin, as in the liver, can be induced (by by ester hydrolysis and/or hydroxylation (phase I
dexamethasone, for example) or inhibited (chloram- metabolic reactions). Unlike mammals, fish are
phenicol, imidazole antifungal agents). The clinical extremely sensitive to pyrethroid toxicity.
significance of altered microsomal enzyme activity in Synthetic pyrethroids (cypermethrin, deltamethrin,
the epidermis has not been established. Metronidazole, fenvalerate, flumethrin, permethrin) are available in a
which is available as a topical gel, inhibits acetaldehyde variety of dosage forms for topical application to
Veterinary Dosage Forms 3971

Table 17 Dosage forms and methods of application of parenteral solutions (for subcutaneous injection), vari-
topical ectoparasiticide preparations to individual species ous oral dosage forms, and topical solutions (Table 19).
Method of The different preparations are specifically designed for
Animal species Dosage form application use in certain animal species. The topical solution of
Cattle Solution Pour on ivermectin or doramectin is applied to cattle by
Liquid concentratea Spray pouron, whereas the topical solution of selamectin is
Ear tag Attach to ears applied to dogs and cats by spot-on. Preparations for
Sheep Liquid concentratea Dip, spray spot-on application contain concentrated solutions of
Solution Spot on, pour on ectoparasiticides and should be applied directly to
Pigs Solution Pour on, spot on
the skin at one (cats) or two (dogs) locations where
Liquid concentratea Spray the animal is unable to ingest the drug by licking.
The back of the neck is one such site. The total dose
Horses Solution Pour on
Liquid concentratea Spray, shampoo
of drug to apply will vary with the animal species
Lotion Dab on (lower dose for cats) and with the range of body weight
(dogs). Selamectin solution, for example, is commer-
Dogs Solution Spot on, spray
and cats (dogs)
cially available in six dose sizes (from 15 to 240 mg).
Collar Surrounding neck The classes of ectoparasiticide and the dosage forms
Dusting power Apply to coat used in cattle, an arbitrarily selected species, are shown
Liquid concentratea Sponge on (dogs) in Table 20. Even though different classes of drug may
Shampoo Wash (dogs) have activity against a similar range of ectoparasites,
a
Liquid concentrates must be appropriately diluted before use on the drugs that belong to the various classes differ
animals. in clinical effectiveness. Quantitative differences in
clinical effectiveness could be attributed to parasite
susceptibility, which would be decreased by the devel-
opment of resistance, and to the combined effect of
domestic animals (Table 18). The appropriate dosage drug access to the site where the parasite is located
form of a pyrethroid is primarily determined by the and persistence of the drug in skin. Both access and
animal species, whereas due consideration is given to persistence are influenced by the physicochemical
the ectoparasites and insects that affect the species. properties of the drug, the formulation of the prep-
Aerosol sprays are more suitable for application to aration, and the method of application to the animal.
dogs than to cats, because cats resent (fearful response) Dermatological preparations containing antibac-
being sprayed and lick the applied preparation from terial agents include aerosol sprays (oxytetracycline
their coat. The combination preparation containing hydrochloride, neomycin sulfate in propylene glycol),
fenvalerate (0.09%), a synthetic pyrethroid, and gels (metronidazole, fusidic acid), creams (gentamicin
diethyltoluamide (9.5%), a cutaneous penetration- sulfate, neomycin sulfate and zinc bacitracin, silver sul-
enhancing substance, has been reported to cause acute fadiazine), ointments (chlortetracycline hydrochloride,
toxicity in cats and occasionally in dogs.[126,127] sodium fusidate, framycetin sulfate and gramicidin,
Macrolide endectocides (avermectins and milbemy- neomycin sulfate, zinc bacitracin, polymyxin B sulfate),
cins) have activity at low concentrations against both and dusting powders (chlortetracycline hydrochloride
internal (nematode) and external (arthropod) para- and benzocaine). The type of preparation influences
sites. Veterinary preparations of avermectins include the stability and release of the active ingredients.

Veterinary–
Virtual
Table 18 Dosage forms of preparations containing synthetic pyrethroids and the methods of topical
application to various species
Dosage form Animal species Method of application
Solution Cattle, sheep, horses, pigs, dogs, cats Pour on
Spot on, spray (dogs)
Liquid concentrate Cattle, sheep, horses, poultry Spray, dip (sheep)
Dusting powder Dogs, cats Apply to coat
Shampoo Dogs Wash
Collar Dogs, cats Surrounding neck
Ear tag Cattle Attach to ears
Synthetic pyrethroids: cypermethrin, deltamethrin, fenvalerate, flumethrin, permethrin.
 3972 Veterinary Dosage Forms

Table 19 Dosage forms and routes of administration of avermectins to various animal species
Dosage form Avermectin Animal species Route of administration
Parenteral
Solution Ivermectin, doramectin Cattle, sheep S.C. injection
Ivermectin pigs, (cats) S.C. injection
Oral
Solution Ivermectin Sheep, goats Drench
Controlled-release ruminal bolus Ivermectin Cattle P.O.
Controlled-release ruminal capsule Ivermectin Sheep P.O.
Premix Ivermectin Pigs Addition to feed
Paste Ivermectin Horses P.O.
Tablet Ivermectin Dogs P.O.
Topical
Solution Ivermectin, doramectin Cattle Pour on
Solution Selamectin Dogs, cats Spot on

Gels and creams are generally easier to apply to animals In the treatment of superficial skin infections, a topi-
but are less occlusive than ointments. Ointments pro- cally applied antibacterial preparation may suffice.
vide a longer duration of drug action, and the occlusive However in severe and deep-seated skin infections,
effect may enhance preparation of the active ingredi- both local and systemic therapy should be applied.
ent(s) to the site of infection. The avoidance of antagonism requires that due con-
The selection of an antibacterial agent should be sideration be given to the mechanisms of action of
based on the clinical diagnosis and, whenever feasible, drugs selected for concurrent use.
in vitro culture and susceptibility testing (when con- Cutaneous mycotic infections caused by filamentous
sidered necessary) of the micro-organisms(s) isolated. or dermatophytic fungi can be treated either topically

Table 20 Representative ectoparasiticides for application to cattle

Ectoparasiticide class
(representative drugs) Spectrum of activity Dosage forms (methods of application)
Organophosphate:
(Phosmet) Mites, lice Solution (pour on)
Warble-fly larvae
Pyrethroid:
(Cypermethrin) Lice, flies Solution (pour on)
(Deltamethrin) Solution (spot on)
Veterinary–

(Fenvalerate) Liquid concentrate (spray)a

Virtual

(Permethrin) Ear tag

Amidine:
(Amitraz) Lice, mites, ticks Liquid concentrate (spray)
Avermectin:
(Ivermectin) Mites, lice, Solution (pour on)
(Doramectin) Horn fly, Parenteral solution
(Abamectin) Warble-fly larvae (S.C. injection);
(Eprinomectin) Controlled-release ruminal
bolus (ivermectin)
Milbemycin:
(Moxidectin) Similar to avermectins Solution (pour on); parenteral
solution (S.C. injection)
a
Liquid concentrates must be appropriately diluted before use.
Veterinary Dosage Forms 3973

or systemically, depending on the location and severity hormones and to show a higher incidence of pharma-
of the skin lesions. Topical antifungal preparations cokinetic interactions with other drugs that undergo
should be formulated to promote penetration and per- microsomal-mediated biotransformation.
sistence of the drug at the site of infection. Drugs that The therapeutic effectiveness of topically applied
may be applied topically include various imidazole corticosteroids is attributed primarily to their anti-
derivatives (clotrimazole, enilconazole, ketoconazole, inflammatory activity. The relative efficacy of topical
miconazole), natamycin, nystatin, and tolnaftate, corticosteroids appears to be in the following order:
whereas orally administered drugs include ketocona- hydrocortisone, prednisolone, betamethasone <
zole, fluconazole, itraconazole, nystatin, and griseoful- hydrocortisone valerate or butyrate, betamethasone
vin. Superficial infections caused by Candida spp. may valerate, triamcinolone acetonide, flucinolone
be treated locally by applying an imidazole derivative acetonide < betamethasone dipropionate, fluocino-
or nystatin. nide. In addition to the nature of the corticosteroid,
In domestic animals and humans, the common fun- its solubility, and, to a lesser extent, the concentration
gal organisms that cause skin infections are Trichophy- used, clinical efficacy is influenced by the formulation
ton mentagrophytes (the usual cause of ringworm in of the preparation. Glucocorticoids appear to have
calves between 2 and 7 months of age), Microsporum greater efficacy when formulated in ointment bases
gypseum, and T. verrucosum (in species other than than in cream or lotion vehicles. This could be attribu-
dogs and cats). Superficial mycotic skin infections are ted to the occlusive effect provided by ointments. The
also caused by T. equinum (horses, cattle, humans), application of an occlusive dressing further enhances
M. nanum (pigs, cattle, humans), and M. canis (dogs penetration and persistence of the steroid (reservoir
and cats). Filamentous fungal keratitis in horses is effect) in the stratum corneum.[112,128]
usually caused by Fusarium spp. Yeasts, primarily
Candida spp., can cause mastitis in cows and metritis
in mares and occasionally inhabit the skin, primarily TRANSDERMAL THERAPEUTIC SYSTEMS
at mucocutaneous junctions, in dogs.
For topical application, individual imidazoles and A transdermal therapeutic system is a rate-controlled
nystatin are formulated as creams, whereas tolnaftate drug-delivery system that, applied to the surface of
and nystatin are available as ointments. Enilconazole the skin, continuously releases the drug at a rate that
is formulated as a liquid concentrate, that must be will provide a desired steady-state plasma concen-
diluted before topical application by wash to horses tration for a specified duration.
and dogs or by spray to cattle. Topically applied enil- The transdermal therapeutic system containing
conazole may be used in conjunction with systemic fentanyl [m (primarily)-opioid agonist], which was
(oral) treatment with griseofulvin. Ketoconazole alone designed to release the drug at a constant rate for
and miconazole nitrate combined with chlorhexidine 72 h, may have application in dogs for the control of
gluconate are commercially available as shampoos postoperative (surgical) pain. Secure placement of the
for dogs. After topical applciation, imidazoles attain transdermal system that releases 50 mg of fentanyl/h
effective concentrations and persist in the outer layers on the dorsal aspect of the thorax of Beagle dogs
of the epidermis, and percutaneous absorption appears (11.4–16.5 kg body weight) provided an average steady-
to be minimal. Natamycin is commercially available as state plasma fentanyl concentration of 1.6 ng/ml, which
a suspension, to be diluted before application by is within the range of plasma concentrations (1–2 ng/ml)
spray to horses or cattle. In addition to spraying horses considered to provide analgesia without producing other
affected with ringworm, all grooming utensils and significant effects.[129,130] A steady-state concentration of

Veterinary–
tackle must be thoroughly cleansed and immersed in fentanyl in plasma was reached at approximately 24 h

Virtual
the natamycin suspension, which should be diluted in after placement of the transdermal system, as would be
plastic or galvanized containers. Natamycin (5% aque- expected because the half-life of fentanyl in Beagles is
ous suspension) is effective in the treatment of filamen- 6 h and was maintained until removal of the system at
tous fungal keratitis, particularly when caused by 72 h. Because the dosing rate (3.7 mg/h  kg) exceeded
Fusarium spp. in horses. the systemic clearance (2.7 mg/h  kg) and the trans-
The selectivity of action of antifungal azoles is dermal bioavailability of fentanyl is 64%, a fraction of
attributed to their greater affinity for fungal than for the released drug either persists in the stratum corneum
mammalian cytochrome P450 enzymes. Because imi- or is metabolized in the epidermis before absorption into
dazole derivatives (clotrimazole, enilconazole, ketoco- the systemic circulation, or both may contribute to
nazole, miconazole) are less selective in their action incomplete systemic availability of the drug. Because of
than are triazoles (itraconazole, fluconazole), the for- the 24 h delay in achieving plasma concentrations within
mer subgroup would be expected to cause greater inter- the analgesia-producing range, either an intravenous
ference with the biosynthesis of endogenous steroid dose (approximately 30 mg/kg) could be administered
 3974 Veterinary Dosage Forms

at the time of placement of the transdermal system or the concentrations of oral flukicides in sheep. J. Vet. Pharma-
system could be securely placed on the animal 12 h col. Ther. 1993, 16, 48–54.
18. Rowlands, D.ap T.; Shepherd, M.T.; Collins, K.R. The
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on an animal, the increased potential for drug inter- tion. J. Controlled. Release 2000, 63, 235–259.
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Bovine Udder and Mastitis; Sandholm, M., Honkanen- tive pharmacokinetics and inversion of (R)-ketoprofen in
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78. Aberg, G.; Ciofalo, V.B.; Pendleton, R.G.; Ray, G.; 99. Foster, R.T.; Jamali, F. High-performance liquid chroma-
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83. Benoit, E.; Soraci, A.; Delatour, P. 6th International Con- sites in nine species. J. Invest. Dermatol. 1990, 95,
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1274–1280. taneous absorption and recovery of parathion in pigs.
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Virtual
 Veterinary Pharmaceuticals: Factors Influencing
Their Development and Use
Marilyn N. Martinez
Laura Hungerford
U.S. Food and Drug Administration, Rockville, Maryland, U.S.A.

Mark G. Papich
Department of Anatomy, Physiological Sciences and Radiology, North Carolina
State University, Raleigh, North Carolina, U.S.A.

INTRODUCTION spectrum of physiological characteristics associated

with the intended market population.
Veterinary pharmaceuticals serve an important role in There are also key differences in the evaluation pro-
preserving and restoring animal health, thereby also cess for human and animal products due to separate
enhancing human well being. Efficient development language in the law and regulations and to human
of safe and effective new animal drugs and continued health considerations for drugs used in food animals.
availability of approved products is essential to main- Unlike human drugs, each pharmaceutical product
tenance of animal health and productivity. For animal receiving U.S. Food and Drug Administration (FDA)
companions, veterinary pharmaceuticals are used to approval for use in an animal species must be codified
treat a range of disease conditions similar to those of [i.e., cited in the U.S. Code of Federal Regulations
human patients. For example, in dogs, drugs are (CFR), listed the 500 series]. The product information
needed to treat conditions such as infection, pain, heart in each CFR citation is summarized in Table 3.
disease, anxiety, and cancer. For poultry, livestock and
aquatic species, therapeutic needs include treatment of
bacterial and parasitic infections, management of MANAGEMENT SETTING AND METHOD
metabolic disorders, productivity enhancers (e.g., OF DRUG ADMINISTRATION
enhancing growth, reproduction, feed efficiency, and
milk production), and control of pain and pyrexia. Products reflect the methods of drug administration
Understanding the needs and constraints of animal most closely aligned to the lifestyle and husbandry
species and the structure and agencies involved in the practices for the targeted animal species. Drugs for
regulatory process is crucial for optimizing animal companion animals share similarities with pediatric
drug development and use. The process leading to treatments because they must be administered by a
approval and marketing of new animal drugs is similar human caretaker. Solid oral dosage forms, such as
to the process for human pharmaceuticals. In fact, the tablets and capsules, are often used for dogs because
majority of animal drugs were initially developed for their mouths can be opened to insert the medication.
potential use in humans. However, veterinary pharma- This may be more troublesome for cats because their
Veterinary–

ceutical development must deal with challenges beyond mouths are more difficult for owners to safely open.
Virtual

the within-species variability experienced in human Medications for dogs can be flavored, administered
studies (Table 1). as chewable tablets, or disguised in a taste treat
Effects of species differences in drug pharmaco- (cheese, for example). However, cats tend to be more
kinetics have been reviewed in a previous volume of discriminating with regard to tastes and consistency
the Encyclopedia of Pharmaceutical Technology for and will refuse medications disguised in food. Many
a wide range of dosage forms. These physiologic differ- liquid medications or, when scored, broken tablets
ences are critical considerations in study design and are so unpalatable to cats that they will salivate and
evaluation during drug development. Variation among resist attempts to administer the drug. Products for
breeds, and individuals within the same species also pet birds and reptiles are often prepared as liquids or
affects treatment outcome (Table 2). Therefore as in powders to add to water or food to limit stress from
clinical trials for human pharmaceuticals, clinical trials animal handling. Because of the problems with medi-
used to support approval of veterinary dosage forms cating some species, various compounding practices
are designed to include animals representing the wide have been used.
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-120026640
3978 Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved.
Veterinary Pharmaceuticals: Factors Influencing Their Development and Use 3979

Table 1 Challenges facing veterinary drug product feed or water, individual treatments (e.g., parenteral
development administration) must be used.

Enormous diversity in size, behavior, metabolic needs, These characteristics have led to specific animal spe-
and lifespan among animal species cies–dosage form combinations among the range of

Species and breed differences in both pharmacokinetic approved veterinary drug products.[1] The United
and toxicity profiles States Pharmacopeia (USP) also has definitions for

A wide spectrum of disease agents that produce these formulations:[2]
different disease manifestations under different
conditions
Liquid dose forms: oral suspensions, solutions,

A range of husbandry practices, which includes a pastes, syrups: These formulations are used across
variety of settings in which animals are kept—ranging all animal species. In some cases, liquid formula-
from animal companions in the home to large tions are added to feed (e.g., added to grain for
livestock operations horses or to moist food for dogs and cats).

Lack of ability to directly communicate with and
Pellets and granules: These are most commonly
educate the animal patient added to or used as feeds for horses and cattle.

Economic constraints Occasionally, this type of medicated feed is used
as a method of drug delivery for swine. When used

Public health concerns
for the treatment of companion animals, pet owners
mix the medications with food. However, this prac-
tice may result in palatability problems and drug
Mass medication of water or feed is an efficient stability concerns. This type of dosage form may
method of delivering medication to a group of animals, also be used to administer drugs to poultry and fish.
such as in a livestock pen. However, treatment of ill
Solid dose forms: tablets, capsules and boluses:
animals may be compromised because the healthiest These dosage forms are most commonly for admin-
animals in the group will out-compete the sick ones istration to dogs and cats. Capsules and some
for access to feed, and sick animals may simply be tablets are swallowed whole. The dosage form is
too anorexic to eat or drink. Therefore, livestock are placed on the back of the pet’s tongue and its
often separated into pens of animals that vary in sever- mouth is held shut, thereby forcing ingestion.
ity of illness. In situations where there is a question of Chewable formulations are often, treat-like formu-
whether or not a sick animal will consume medicated lations and may be swallowed whole, chewed, or

Table 2 Breed idiosyncrasies affecting drug pharmacokinetics

Group/species Subgroup/breed Idiosyncrasy References
Canine Greyhound Decreased body fat results in decreased volume of distribution [3,4]
of fat soluble drugs such as anesthetics and thiobarbiturates.
Lower cytochrome P450 (CYP2b11) levels and hydroxylase [5,6]
activity decreases rate of metabolism for some compounds,
leading to longer T1/2.
Beagles Higher hepatic clearance for some compounds (e.g., phenobarbital, [7]
which is twice as fast in Beagles as in mixed breeds)

Veterinary–
Virtual
Large breeds (e.g., Higher paracellular intestinal permeability as compared to [8]
Giant Schnauzers smaller breed dogs.
and Great Danes) Lower gastrointestinal mass as compared to small breed dogs, [9–11]
but no difference in GI transit time.
Collies, Australian Autosomal recessive defect in MDR1 gene, leading to defect in [12–16]
Shepherds, Old English P-gp. This has caused a defective blood brain barrier, resulting in
Sheepdogs, and some ivermectin toxicity (depression, coma, and death), loperamide
related breeds toxicity (sedation), and adverse effects from chemotherapeutic drugs.
Approximately 30% of Collie dogs are believed to be affected.
Equidae Donkeys vs. horse Difference in metabolism of NSAIDs. [17–19]
vs. ponies vs. mules
Cattle Aberdeen Angus Delayed and reduced moxidectin topical absorption in Angus vs. [20]
vs. Holstein Holstein, which may explain variability in drug effectiveness.
 3980 Veterinary Pharmaceuticals: Factors Influencing Their Development and Use

Table 3 Product information contained within the CFR The manufacture of a Type B medicated feed

Name of the drug requires that the feed mill has an approved

Name of the drug sponsor medicated feed mill license.
– Type C: This is intended as the complete feed

The approved dose, route of administration
for the animal, added to the top of the usual

The approved indication (disease and target rations, or offered ‘‘free-choice’’ in conjunction
animal species)
with other animal feed. It is manufactured by

Withdrawal time and the tolerance (for products to diluting a Type A medicated article or a Type
be used in food-producing animals). Information regard- B medicated feed with feed and may be further
ing the marker residue and target tissue can be found in
diluted to form another Type C medicated feed.
the freedom of information summaries that are posted
The manufacturer of a Type C medicated feed
on the CVM website (http://www.fda.gov/cvm/efoi/
foidocs.htm) must have an approved medicated feed mill
license if that feed is produced from a Type A
medicated article containing drugs associated
with a withdrawal period at its lowest use level.
mashed and placed on food. Oral boluses are large,
oblong shaped, formulated for cattle or horses, and
Injectable products: These products are used
often contain >1000 mg drug substance. Cattle are across all animal species (including fish). Products
medicated via a ‘‘balling gun’’ or dosing device may be formulated for subcutaneous (SC), intra-
that, when inserted into the mouth of restrained muscular (IM), or intravenous (IV) injection. For
cattle, can deliver the bolus to the back of the ton- some species (e.g., cattle), a large proportion of
gue whereupon it is swallowed. While large boluses the approved therapeutic drug products are parent-
have been available for horses (e.g., 1 g phenylbuta- erally administered. Cattle are often dosed while
zone tablets), these are generally crushed, mixed restrained in chutes. While this method of adminis-
into a thick paste (with molasses or corn syrup for tration insures that each animal receives the pre-
example), and administered with a dosing syringe. scribed dose, it can be associated with greater

Drenches: These liquid formulations, used pri- animal stress and higher costs of administration
marily in horses and ruminants, are poured into than other routes of administration (e.g., pour-on
the back of the mouth or may be administered products). Accordingly, sponsors of injectable pro-
using gastric or nasogastric tubes. ducts often focus on developing injectable formula-

Medicated water: This method for treating groups tions that are effective with the fewest possible
of animals is particularly valuable for maximizing number of doses per animal. Injectable formula-
the likelihood of drug intake during a disease out- tions also present with concerns regarding injection
break (animals will often drink fluids even when site irritation and injection site residues. Thus, sep-
anorexic). For this reason, it is often used to treat arate injection site tests for drug residues are
swine, cattle, poultry, and fish. needed, and may result in a ‘‘trim out’’ statement

Medicated feeds: These are most frequently used as in the approved product label (e.g., oxytetracycline
a method of drug administration to cattle, swine, for use in cattle[21,22]). There is also the concern
horse, poultry, and fish. Three types of medicated about carcass quality for IM injections. Bruises
feeds are defined in 21 CFR 558.3: and blemishes in cuts of meat will lower its quality,
even if there are no human food safety (HFS) con-
– Type A: This medicated article is intended cerns resulting from drug residues. Therefore, some
Veterinary–

solely for use in the manufacture of another sponsors have filed supplemental applications to
Virtual

Type A medicated article or a Type B or Type support the approval of an SC route for a product
C medicated feed. It consists of an animal previously approved for IM injection (the hide is
drug(s), with or without carrier (e.g., calcium removed at the time of slaughter, thereby removing
carbonate, rice hull, corn gluten) and with or any blemishes associated with a SC injection site).
without inactive ingredients. The manufacture Alternatively, some of the more recently previously
of a Type A medicated article requires an appli- approved IM products are labeled for injection into
cation approved under 21 CFR x 514.105. the neck muscle because when the head is removed
– Type B: This medicated feed is intended solely at the time of slaughter, the injection site is also
for the manufacture of other Type B or Type removed. With regard to IV administration,
C medicated feeds. It contains no less than problems confronting veterinary use parallel those
25% nutrients per unit weight. It is manufac- associated with human IV injections.
tured by diluting a Type A medicated article
Topical products: Examples exist for most animal
or another Type B medicated feed with food. species. Transdermal formulations that achieve
Veterinary Pharmaceuticals: Factors Influencing Their Development and Use 3981

quantifiable systemic drug concentrations are ECONOMIC CONSIDERATIONS

highly lipophilic, penetrate the skin, and achieve a
sustained effect. These drugs are typically adminis- Economics plays an important role in animal treat-
tered in a vehicle that enhances absorption. Their ment decisions. While, for owners of a beloved dog
lipophilicity and slow elimination produce a sus- or cat, cost considerations may be less important, most
tained effect after a single application. One such animal health care is not covered by insurance. There-
product is fipronil (FrontlineÕ), which is seques- fore, owners are limited to the resources they can
tered by the hair follicles and sebum of the skin. devote to the treatment of a pet. Costs are even more
The drug is gradually released from these sites, critical for farmers working within narrow profit mar-
allowing for monthly application for the control gins. In addition to the costs associated with the drug
of fleas and ticks. Other successful transdermal itself, farmers need to consider costs associated with
formulations used for control of parasites are animal handling and the potential impact of handling-
imidacloprid (AdvantageÕ) and selamectin induced stress on animal growth and feed efficiency
Õ
(Revolution ). Transdermal drugs for cattle include (the ‘‘chute factor’’). Prolonged withdrawal periods
the pour-on antiparasitic drug, ivermectin. Many can adversely affect the quality of meat and can
pour-on insecticides and powders also are available. increase farmer costs because of the need to retain ani-
Although a drug may be administered topically, mals on the farm for a longer period of time. Even rela-
animal-licking behavior can result in a portion of tively small costs can have an impact with large groups
the dose being swallowed.[23] Potential conse- of animals and slim profit margins.
quences of ingesting a portion of the topical dose
need to be considered prior to product approval.
Some topical products have minimal systemic
absorption but are associated with therapeutic ANTIMICROBIAL RESISTANCE
claims. An example is otic preparations (e.g.,
MometamaxÕ, NADA #141-177). Topically applied Resistance of pathogens to antimicrobial treatment is a
ectoparasiticides that are not systemically absorbed major concern in veterinary medicine, just as in human
are regulated by the Environmental Protection medicine. Antimicrobial resistance negatively impacts
Agency (EPA) rather than by the FDA.[24] both the current use and future development of phar-

Medicated collars and ear tags: Drugs with anti- maceuticals for animals. For sick animals, anti-
parasitic action, for example to ward off biting flies, microbial resistance directly impairs the success of
ticks, and lice, have been formulated into ear tags treatment to control disease. This can affect the prog-
for cattle, and collars for dogs and cats. These nosis and suffering for an individual animal and often
devices, with active ingredients impregnated into the productivity, survival, and economic returns for an
the plastic, are effective by gradually releasing the entire herd. Resistance may make it difficult to find
drug to repel insects. These drugs can occasionally effective, already approved drugs as active controls
produce toxic systemic effects, which have been for preapproval clinical trials. The ability of resistant
observed in dogs. Similar to other topical ectopara- micro-organisms to move between humans and
sitic preparations, these products are regulated by animals raises serious public health concerns.
the EPA and not FDA. Resistance among endoparasites and ectoparasites,

Intramammary preparations and vaginal inserts: such as those that cause malaria and schistosomiasis
These products are generally formulated for use in in humans, or for animal nematodes, is a well-
cattle. Intramammary products are intended for recognized problem. In veterinary medicine, integrated

Veterinary–
the treatment of mastitis-related claims (e.g., parasite control and rotation of different parasiticides

Virtual
PirsueÕ, NADA 141-036). Vaginal inserts are gener- has been used to slow development of resistance. For
ally used for the estrus regulation/synchronization many parasites, current agents are very effective.[25]
(e.g., Eazi-Breed CIDRÕ, NADA #141-200). Due to host specificity, there has been little problem
with zoonotic human disease caused by resistant
animal parasites.
COMPLIANCE AND COMMUNICATION Resistance to antiviral agents has been of lesser con-
cern in veterinary than human medicine and data are
In human medicine, the majority of drugs are formu- limited on the prevalence of resistance among animal
lated as oral dosage forms. Concerns of user non- viral pathogens. Antiviral treatment is not widely used
compliance have led to the development of sustained in animals, often because of cost. Management has
release formulations. While user noncompliance is also focused on prevention of animal disease through vacci-
a concern in veterinary medicine, the focus is on the pet nation or eradication of diseases from herds and geo-
owner administering the drug. graphic regions. Most animal viral pathogens are
 3982 Veterinary Pharmaceuticals: Factors Influencing Their Development and Use

non-zoonotic, with notable exceptions (avian influenza, Preventing both occurrence of resistant organisms in
Nipah virus). Although poultry in Asia had not been food and transmission of resistant organisms between
treated with antiviral agents and more than 60 million species and individuals are crucial to controlling this
were destroyed during the initial H5N1 avian influenza health risk.[42]
outbreak in 2003, viral samples from sick humans were FDA provides a website with public information on
resistant to amantadine and rimantadine.[26] this issue,[43] as well as specific guidance for indus-
Resistance to antibacterial agents strongly affects try.[44,45] These documents describe a process for decid-
animal drug development and use. Emergence of resist- ing if a proposed antimicrobial use in food animals
ance is a complex ecological phenomenon involving would likely lead to resistance that would negatively
genetic change, microbial selection, dissemination of impact human health. It outlines possible methods
resistance in the host and population, and persistence for: ranking antimicrobials based on their importance
of resistant phenotypes.[27–32] Antimicrobial use can for human use; conducting a qualitative risk assess-
decrease prevalence of pathogens, but it can also exert ment for a proposed animal antimicrobial use; and
selection pressure for resistant organisms. Increased managing risks identified in the assessment.
prevalence of resistant bacteria in animals poses a In addition to regulation, FDA has been actively
potential risk to humans as well as animals. Contro- involved in research, monitoring, and setting stan-
versy surrounds the optimal measures to preserve the dards. The Center for Veterinary Medicine research
effectiveness of these treatments for humans and ani- program includes projects to evaluate effects of anti-
mals.[33,34] Principles for judicious use of antimicro- microbial use in animals on efficacy against pathogens,
bials have been developed and continue to be refined changes in the environmental microbial ecology, and
to combat this problem in domestic animals.[35,36] development of antimicrobial resistance in pathogenic
In several countries, antibacterial use has been limited and commensal micro-organisms. The National Anti-
in animals as a public health precaution, particularly microbial Resistance Monitoring System (NARMS)
species that can serve as a source of foodborne was established as a collaborative effort between the
disease.[29] FDA, USDA, and the Centers for Disease Control
In companion animals, the bacterial infections most (CDC).[46] This program is responsible for monitoring
frequently requiring treatment involve the skin, changes in the susceptibilities of human and animal
wounds, ears, respiratory tract, and urinary tract.[37] enteric bacteria such as E. coli, Salmonella spp.,
Prevalence of resistant bacteria is difficult to assess in Enterococcus spp., and Campylobacter spp. It can
pets. Since empirical treatment is common, there is lim- serve as a resource for antimicrobial risk management
ited information on trends in resistance, and almost no plans developed during approval of animal antimicro-
data are available on the magnitude of antimicrobial bials. Its objectives include:
use in these species. Increasing resistance among E. coli
and Staphylococcus isolates from pets has been
reported from studies in a number of countries.[38]
Providing descriptive data on the extent and
Methacillin-resistant Staphylococcus aureus (MRSA) temporal trends of antimicrobial susceptibility in
have been isolated from dogs[37] and horses,[39] Salmonella and other enteric organisms from
although animal outbreaks have generally been traced human and animal populations.
to infected humans.[38]
Identifying the emergence of microbial resistance in
Among food animals, antimicrobials are used to humans and animals.
treat diseased animals, to prevent or control disease
Providing timely information to veterinarians and
outbreaks, and in the feed to enhance performance. physicians.
Veterinary–

Enteritis, mastitis, and respiratory disease are the most
Prolonging the lifespan of approved drugs by pro-
Virtual

frequently treated infections.[40] Decreased sensitivity moting prudent and judicious use of antimicrobial
of food animal pathogens has been reported for com- agents.
monly used antimicrobials, such as penicillins, strepto-
mycins, tetracyclines, and sulfonamides.[37] Resistance
impairs both success and cost effectiveness of animal Each year, Salmonella and other enteric organisms
health management. Of further concern is potential isolates are obtained from animal and human sources
foodborne transmission of resistant bacteria to and their susceptibilities are tested to approximately
humans, especially with the large-scale transport and 17 drugs of human importance. The human samples
distribution of food products. This includes increased are collected from sick people. The animal isolates
prevalence of resistance among pathogens, such as come from healthy farm animals, animal clinical
Salmonella and Campylobacter,[33,40] and resistance in specimens, carcasses of food animals at slaughter,
normal animal flora that subsequently transfer resistance ground products at processing plants, and retail meat
to opportunistic pathogens in the human gut.[41] samples collected from grocery stores and shops that
Veterinary Pharmaceuticals: Factors Influencing Their Development and Use 3983

sell meat to the public.[47] Animal-origin isolates are have not been available. The VAST is comprised of
collected from sites across the United States and are microbiologists, veterinary clinicians, veterinary phar-
sent to the Antimicrobial Resistance Research Unit macologists, representatives from industry and FDA,
(ARRU) of USDA in Athens, Georgia, for suscepti- and advisors.
bility testing. An example of the results of these tests In 2002, the committee has published an M31-A
from veterinary isolates (1997–2003) is provided in document that provides performance standards
Table 4.[48] for susceptibility testing, as well as a document for
Among other measures to insure judicious veterin- in vitro susceptibility testing criteria and quality con-
ary drug use, the National Committee for Clinical trol parameters for bacteria isolated from animals
Laboratory Standards[49] (whose name was officially (available by ordering through CLSI). The CLSI docu-
changed to the Clinical Laboratory Standards Insti- ments provide veterinary diagnostic laboratories with
tute, CLSI, in January, 2005[50]), Committee on Micro- recommended standardized antimicrobial suscepti-
biology, Subcommittee on Veterinary Antimicrobial bility test methods for bacteria, criteria for quality con-
Susceptibility Testing (VAST) was established to pro- trol, a list of drugs considered for routine testing in
vide public standards of veterinary-specific interpretive veterinary microbiology laboratories, and interpretive
criteria for the susceptibility testing of bacterial patho- criteria (determination of resistant vs. susceptible)
gens infecting animals. These interpretive criteria are relative to the clinical use of the agent. Data used by
used to establish breakpoints defining ‘‘resistant,’’ CLSI to interpret susceptibility breakpoints include:
‘‘sensitive,’’ and ‘‘intermediate’’ categories for drug target species pharmacokinetics, pharmacodynamic
susceptibility. The committee has worked to update parameters, efficacy information, and the population
standards, develop standards for new drugs, and evalu- susceptibility distributions of clinically relevant
ate older drugs for which veterinary-specific standards pathogens.

Table 4 National antimicrobial resistance monitoring system animal isolates (chicken slaughtera isolates)
Percent resistance

Antimicrobial 1997 1998 1999 2000 2001 2002 2003b

(number of isolates) (n ¼ 214) (n ¼ 562) (n ¼ 1438) (n ¼ 1173) (n ¼ 1307) (n ¼ 1500) (n ¼ 705)
Amikacin 0 0 0 0 0 0 0
Amoxicillin/ 0.5 2.0 4.9 7.3 4.5 10.2 7.8
clauvalanic acid
Ampicillin 11.7 13.0 12.4 13.0 9.4 14.3 12.2
Apramycin 0 0.2 0.1 0.7 0.6 N/A N/A
Cefoxitin N/A N/A N/A 7.2 4.1 8.7 6.0
Ceftiofur 0.5 2.0 5.2 7.6 4.1 10.2 7.8
Ceftriaxone 0 0.5 0 0.1 0 0.3 0.1
Cephalothin 1.4 4.4 5.8 7.8 4.7 10.5 8.4
Chloramphenicol 2.3 2.8 1.8 4.6 2.5 2.4 1.6
Ciprofloxacin 0 0 0 0 0 0 0.1

Veterinary–
Gentamicin 17.8 15.5 10.4 14.9 7.9 5.5 6.2

Virtual
Imipenem N/A N/A N/A N/A 0 N/A N/A
Kanamycin 2.3 3.2 1.2 4.0 2.4 2.0 2.7
Nalidixic Acid 0.0 0.2 0.2 0.5 0 0.8 0.7
Streptomycin 24.3 27.8 27.5 28.6 21.0 22.9 21.0
Sulfamethoxazole 24.8 23.8 15.9 18.4 11.8 8.9 10.2
Tetracycline 20.6 20.5 25.0 26.3 21.9 24.9 27.1
c
Ticarcillin 11.7 11.7 N/A N/A N/A N/A N/A
Trimethoprim/ 0.5 1.4 1.1 0.4 0.5 0.8 0.6
sulfamethoxazole
a
Isolates obtained from slaughter (carcass swabs) and processing plants (ground product).
b
2003 Preliminary data.
c
In 1999 Florfenicol replaced ticarcillin. No results are listed for Florfenicol due to lack of interpretive criteria for this antimicrobial.
 3984 Veterinary Pharmaceuticals: Factors Influencing Their Development and Use

U.S. REGULATION OF NEW ANIMAL DRUGS U.S. New Animal Drug Applications (NADA)

The Center for Veterinary Medicine (CVM) of the U.S. Substantial evidence of effectiveness
FDA regulates the manufacture and distribution of
food additives and drugs that will be given to animals. 21 CFR x 514.4(b) (2) requires that a sponsor demon-
These include animals from which human foods are strate that its new animal drug is effective for each
derived, as well as food additives and drugs for pet proposed intended use and the associated conditions
(or companion) animals. Veterinary biologics (vaccines, of use. This includes the dose or dose range, frequency,
bacterins, diagnostics, etc. which are used to prevent, duration, timing (e.g., in relation to the onset of clinical
treat, or diagnose animal diseases) are regulated by the signs), animal species, age, gender, class, and breed of
U.S. Department of Agriculture, Center for Veterinary animal for which the new animal drug is intended for
Biologics.[51,52] use. Subsequent to the enactment of the Animal Drug
According to the Animal Health Institute’s 2002 Availability Act of 1996 (ADAA),[56] the specific num-
Market Sales Report, the sale of animal health pro- ber and types of adequate and well-controlled studies
ducts in the United States for 2002 totaled $4.5 billion, for providing substantial evidence of effectiveness
representing an increase of 1.5% compared to 2001.[53] depends upon the number of intended uses, how nar-
In contrast, the total U.S. pharmaceutical sales—the rowly or broadly each intended use is defined, and
world’s largest market for prescription drugs, grew upon the conditions of use in the proposed labeling.
12% in 2002 to reach $219.2 billion.[54] The economic When a proposed indication is associated with a dose
constraints associated with veterinary product devel- range, substantial evidence of effectiveness is demon-
opment necessitates that regulatory decisions for ani- strated for the lowest proposed dose. The highest dose
mal health products be based on a target animal within the dose range is contingent upon a demon-
safety (TAS) and clinical effectiveness database that stration of TAS.[57] Sponsors need to justify the
is markedly smaller than that associated with new drug intended dose. The basis of this justification can
applications for human pharmaceuticals. The legal include pharmacokinetic/pharmacodynamic relation-
requirements associated with veterinary drug product ships, literature, and laboratory tests.[56]
applications are detailed in the CFR 21 CFR x 514.1 Typically, effectiveness studies contain at least 100
and are discussed elsewhere.[55] Technical sections clinical cases per indication (far fewer than the thou-
included in a veterinary drug application are provided sands of subjects included in Phase III human effec-
in Table 5. tiveness studies).[58] Although the ADAA allows for
For a veterinary drug application approved on or the requirement of substantial evidence of effectiveness
after July 1, 1975, after an approval has been published to be met with one well-controlled clinical field trial,
in the Federal Register, a Freedom of Information independent substantiation continues to be necessary.
(FOI) summary must be prepared that summarizes
the safety and effectiveness data and the information Target animal safety
submitted with or incorporated by reference in the
application file [21 CFR x 514.11(e) (2) (b) (ii)]. Infor- 21 CFR x 514.1(b) (8) (i) requires ‘‘adequate tests by all
mation pertaining to veterinary FOI summaries can be methods reasonably applicable to show whether or not
found on the CVM website (http://www.fda.gov/ the new animal drug is safe and effective for use as
cvm/efoi/efoi.html). suggested in the proposed labeling.’’ As with human
pharmaceuticals, published literature and preliminary
studies (including pharmacokinetics and pharmaco-
Veterinary–

dynamics, extrapolation from toxicity testing in lab-

Virtual

Table 5 Technical sections associated with veterinary drug

oratory animals, and observations from clinical trials)
product applications
are considered in the safety evaluation. But for new

Chemistry, manufacturing, and controls section animal drugs, safety is also directly assessed through

Effectiveness laboratory (overdose) studies in the target species.

Target animal safety The specific TAS information needed for a product

Human food safety depends on factors such as proposed usage regimen
and dose, type of drug, chemistry and manufacturing

Environmental impact
considerations, claims, previous use history, and ani-

Labeling mal species including class and breed. Studies generally

‘‘All other information,’’ which includes, but is compare control animals with small numbers of ani-
not limited to, any information derived from other mals of the target species treated with multiples of both
marketing (domestic or foreign) and favorable and the recommended dose and duration of administration
unfavorable reports in the scientific literature
of the product. Studies should be well designed and
Veterinary Pharmaceuticals: Factors Influencing Their Development and Use 3985

conducted in compliance with Good Laboratory Prac- compound, its metabolites, and any other substances
tice (GLP) standards. Appropriate observations, physi- formed in or on food because of the sponsored com-
cal examinations, clinical pathology tests (hematology, pound’s use.’’[62] These residues are the subject of all
blood chemistry, urinalysis, fecal analysis, etc.), nec- HFS evaluations. Toxicology tests are designed to deter-
ropsy, and histopathology should be conducted to mine the dose at which the compound produces an
identify possible adverse effects. Additional specialized adverse effect and a dose that produces no observed
studies, such as evaluation of injection sites, topical effect (NOEL). If the drug is not a carcinogen, the
administration sites, reproductive safety and mammary NOEL of the most sensitive effect in the most sensitive
gland safety, may be needed depending on the con- laboratory species, divided by a safety factor, is used
ditions of use and the characteristics of the drug. There to determine the human acceptable daily intake (ADI)
may also be species-specific study requirements based for that drug. The ADI is estimated as:
upon unique physiology and or husbandry practices.
For example in fish, TAS can change as a function of ADI ðmg=kg=dayÞ
water temperature, ionic strength and composition, ¼ ½ðNOEL ðmg=kgÞÞ=ðSafety Factor ðday1 ÞÞ
and life stage.[59]
FDA is helping to develop internationally accept- The safety factor acts as a buffer, protecting against
able, harmonized guidelines for conducting TAS studies. errors in interspecies extrapolations or human popu-
This will benefit animal welfare and decrease drug devel- lation predictions. Within the World Health Organiza-
opment costs, by reducing redundancy in animal testing tion (WHO),[63] a safety factor of 100 units per day is
for different countries, while continuing to assure the applied when the NOEL is derived from a long-term ani-
safety of new animal drugs. mal study. It is based on an assumption that humans
may be up to 10 times more sensitive than the test ani-
Human food safety mal used and that there may be up to a tenfold range
of sensitivity within the human population. Higher
By the general safety provisions of Sections 409, 512, safety factors (e.g., 200, 500, 1000, and 2000) have been
and 706 of the Federal Food, Drug, and Cosmetic applied when incomplete or inadequate data are submit-
Act,[60] FDA must determine whether each food addi- ted (e.g., too few animals) or when irreversible or terato-
tive, new animal drug, or color additive proposed for genic and carcinogenic effects have been observed.[63]
use in food-producing animals is safe for those animals Average consumption values (grams consumed) for
and whether the edible products derived from treated edible muscle and organ tissue are described in Table 7.
animals are safe for human consumption. Accordingly, These values are factored into the estimate of a safe
the sponsor must demonstrate reasonable certainty of concentration. The safe concentration is, by definition,
no harm to human health for all drug products the amount of residue that can be eaten in any edible
intended for use in food-producing animals. tissue each day for an entire lifetime without exposing
The components of a HFS package are provided in the consumer to residues in excess of the ADI.[64] To
Table 6.[61] estimate the safe concentration, FDA considers the
A residue is ‘‘any compound present in the edible ADI, the weight in kilogram of an average adult
tissues of the target animal which results from the use human (60 kg), and the amount of the product that
of the sponsored compound, including the sponsored may be consumed in grams per day.

Safe concentration ðmg=gÞ

ADI ðmg=kg=dayÞ  60kg

Veterinary–
Table 6 Information comprising the HFS technical section ¼

Virtual

A toxicological assessment grams consumed per day

The determination of an ADI A drug may be intended for use in multiple animal

Determination of a safe concentration of drug species (e.g., where residues may be present in eggs,
residues within the target animal species

Development and validation of an analytical method
that serves as the regulatory method for determining the Table 7 Consumption values
withdrawal time and for assessing whether or not Edible product Grams consumed
animal-derived products contain residue concentration
in excess of the established limit (violative residues of Muscle 300
meats and animal-derived food products] Liver 100

Establishment of a tolerance Kidney 50

Establishment of a withdrawal time Fat 50
 3986 Veterinary Pharmaceuticals: Factors Influencing Their Development and Use

milk, and meat) or may enter humans via other routes milk discard times are established on the basis of some-
(e.g., pesticides on crops). In these cases, the ADI will what different assumptions.[61]
need to be partitioned across the various edible foods To determine the withdrawal time, the U.S. FDA
such that if all are summed daily, the sum of all those recommends the use of regression methods because it
sources does not exceed the ADI for that compound. allows sponsors to obtain information across a wide
For this reason, many drug sponsors may elect to range of potential time points. The necessary confi-
request that a safe concentration be used that is less dence is achieved by the use of tolerance limits rather
than 100% of that which would be allowed by the than confidence (prediction) limits. While confidence
ADI established for that compound. limits enclose a range of possible values for a popu-
A tolerance (the FDA maximum concentration of lation parameter, tolerance limits cover, with a set
the marker residues within the target tissue) or the degree of certainty, a specified percentage of the indivi-
European version of a tolerance, the maximum residue duals within a given population. Accordingly, toler-
limit (MRL) is determined based on this marker resi- ance intervals are be wider than confidence intervals
due.[65] Selection of a marker residue, target tissue, (Fig. 3).[66] This increased width is achieved through
and tolerance is graphically described in Figs. 1 and the use of the noncentral t distribution.[67] Ultimately,
2. The marker residue (the residue that monitors the the regulatory objective of the U.S. FDA procedure is
depletion of total residues in a tissue) is in a known to predict a time when we can be 95% certain that the
relationship with the total radiolabel drug residues tissue residue concentrations in 99% of the animal
such that when the concentration of the marker residue population receiving the drug product (when dosed
in the target tissue is less than the target tissue toler- in accordance to the approved product label) will be
ance, total residues in all of the edible tissues are less at or below tolerance. Concentrations of the marker
than their respective safe concentrations. The marker residue must be measured with an FDA-approved ana-
residue is generally the edible tissue from which the lytical method (the regulatory method). This is the
residues most slowly deplete. same method that is used by validated analytical
The withdrawal time (the time between the last laboratories during inspections, insuring the absence
administered dose and when the drug residues drop of violative residues in marketed human food products
below the safe concentration) is generally established derived from animal sources.
by obtaining the target tissues of 20 animals, with five In addition, Section 512 of the Act has been inter-
animals being sampled at each of four evenly distribu- preted to include the need to confirm that the level of
ted time points. The assumptions associated with the antimicrobial residues present in food have no clini-
statistical analysis are that the measurements are inde- cally relevant effect on the human intestinal microflora.
pendent, the depletion of the natural logarithm (ln) Accordingly, for antimicrobial compounds, antimicro-
concentrations is linear with time, and that the mea- bial-specific issues are considered along with tra-
sured ln concentrations are normally distributed (hav- ditional toxicological studies and additional testing
ing a constant variance over time). Since repeated milk may be necessary.[68] CVM has recently published a
samples are obtained from an individual lactating cow, guidance document that serves as an initial step in

10,000.0
tissue expressed in [ ] units (e.g., PPB)
Total residue levels in various edible

TL= Largest value among TT1,TT2 or TT3

1,000.0
= Target tissue
Veterinary–
Virtual

100.0
Tissue 1

10.0 Safe
Tissue 2 concentration
Tissue 3

1.0

Fig. 1 Total residue depletion curves used

0.1 to select the target tissue, e.g., muscle, liver,
kidney, and fat. (From Ref.[66], reprinted
with permission from Dairy, Food and
0.01
0 1 2 3 4 5 6 7 Environmental sanitation. Copyright held
Time by the International Association for Food
(Appropriate units, i.e., hours, days, etc.) protection.)
Veterinary Pharmaceuticals: Factors Influencing Their Development and Use 3987

5.0 If P2 is selected to be marker residue then the

(In appropriate units, i.e., PPM, PPB, etc.) 4.0 level of marker residue that the regulatory assay
must be capable of measuring is Rm.
3.0
Residues in target tissues

Total residue in target tissue,

2.0 i.e., P0 + P1 + P2
Safe
concentration

1.0
P2
Fig. 2 Selection of a marker from among the indi-
0.5 Rm P1 vidual residues (metabolites) within the target tissue.
P0 The Rm (tolerance) represents the concentration of
TL from the marker residue when the total residue achieves
figure 1 its safe concentration. (From Ref.[66], reprinted with
0.2 permission from Dairy, Food and Environmental
0 1 2 3 4 5
Time sanitation. Copyright held by the International
(In appropriate units) Association for Food protection.)

developing harmonized technical guidance for appro- determine if the drug is approvable under specific risk
val of therapeutic antimicrobial veterinary medicinal management conditions. FDA is concerned about a
products intended for use in food-producing animals range of deleterious effects that antimicrobial resistant
with regard to characterization of antimicrobial bacteria may have on human health, including an over-
resistance selection in bacteria of human health riding concern about the potential for a decrease or
concern in the European Union, Japan, and the United loss of effectiveness of antimicrobial drugs in humans
States.[69] as a consequence of human exposure to resistant
Human food safety is also evaluated with respect to bacteria through ingestion of animal derived food pro-
the potential microbiological effects of antimicrobial ducts. Due to the difficulties associated with measuring
new animal drugs on foodborne bacteria of human these effects, regulatory decisions are made on the
health concern. As shown in Fig. 4, the FDA/CVM basis of the probability of the occurrence of the hazard.
Guidance #152[70] provides a qualitative risk assess-
ment approach for examining this issue. This integrat-
ive method of generating a risk assessment provides an U.S. Abbreviated New Animal Drug
overall conclusion regarding the potential risk to Applications (ANADA)
human health of the proposed use of the antimicrobial
new animal drug. FDA/CVM then uses the resulting In 1984, President Reagan signed into law the Drug
risk estimation ranking, along with other data and Price Competition and Patent Term Restoration Act
information submitted in support of the NADA, to (Waxman Hatch Act). This legislation created a system

4.0
3.5
Log residue [ ] in PPM, PPB, etc.

Veterinary–
3.0 99% Statistical tolerance limit

Virtual
with 95% confidence
2.5
2.0
1.5 Mean of
residue data
1.0
0.5 Fig. 3 Determination of the withdrawal time
0.0 Tolerance value from the 95% confidence bound on the 99%
statistical tolerance limit on the residue deple-
– 0.5
tion data rounded to the next day. (From
–1 Ref.[66], reprinted with permission from Dairy,
0 1 2 3 4 5 6 7 8 Food and Environmental sanitation. Copyright
Days Withdrawal time held by the International Association for Food
or (WDT) protection.)
 3988 Veterinary Pharmaceuticals: Factors Influencing Their Development and Use

Hazard characterizaton

Qualitative risk assessment

Probability that resistant bacteria are present in

Release
target animal as a consequence of drug use
assessment
(rank as High, Medium, or Low)

Probability for humans to ingest bacteria in question

Exposure from the relevant food commodity
assessment (rank as High, Medium, or Low)

Consequence Probability for humans exposure to resistant

assessment bacteria results in an adverse health consequence
(rank Important, Highly important, or Critically important)

Overall risk estimate: Integration of

release, exposure and consequence
Risk estimation
assessments
(rank as High, Medium, or Low)

Fig. 4 Components of a qualitative antimicrobial resistance risk assessment. (From Ref.[70].)

for the review and approval of generic versions of post- blood levels are measured under steady-state con-
1962 human ‘‘pioneer’’ drug products through the use ditions, the AUC is measured over a single dosing
of abbreviated new drug applications (ANDAs). In interval (AUC 0–t). Statistically, CVM advocates use
1988, Congress enacted the Generic Animal Drug of the 90% confidence intervals (the 2 one-sided test
Patent Term Restoration Act (GADPTRA) to create procedure originally described by Schuirrman[74]).
a similar system for animal drug products. ANADAs The limits necessary to define bioequivalence are gen-
contain all eight technical sections applicable NADAs, erally 0.80–1.20 (untransformed data, based upon
but the information needed to fulfill these requirements treatment differences and expressed relative to the ref-
is different. For example, while every application erence mean) and 0.80–1.25 (Ln-transformed data,
must contain either an environmental assessment, expressed as the ratio of the test/reference means).
sponsors of an ANADA generally file a request for a Alternatively, one can use statistical methods described
categorical exclusion.[71] Technical sections for TAS in CDER guidance documents.[75] Prior to initiating
and effectiveness are obtained through the demon- studies, CVM protocol concurrence is always strongly
stration of product bioequivalence. encouraged.
Bioequivalence studies (i.e., blood level, pharmaco- CVM has concluded that tissue residue depletion of
Veterinary–

logic end-point, and clinical end-point studies) and generic products are not adequately addressed through
Virtual

tissue residue depletion studies are conducted in bioequivalence studies unless the pioneer product is
accordance with GLP regulations.[72] When absorption approved with a zero withdrawal time (i.e., the animal
of the drug is sufficient to enable the quantification of may be sacrificed for human consumption at any time
drug concentrations in the blood (or other appropriate after drug administration). Therefore, sponsors of
biological fluid or tissue) and if systemic absorption is ANADAs for drug products for food-producing ani-
relevant to drug action, a blood (or other biological mals are asked to conduct bioequivalence and tissue
fluid or tissue) level bioequivalence study should be residue studies unless blood concentrations of the
conducted. When using blood level studies, the pivotal active drug can be measured out to the withdrawal
parameters supporting product comparability are the time of the pioneer product. A tissue residue study
area under the concentration vs. time curve (AUC) should generally accompany clinical end-point and
measured from time zero to the last quantifiable drug pharmacologic end-point bioequivalence studies.[76]
concentration (for a single dose study), and the The holder of the generic drug product application
maximum observed concentration (Cmax).[73] When uses the existing tolerance, marker residue, target
Veterinary Pharmaceuticals: Factors Influencing Their Development and Use 3989

tissue, and analytical method (as contained within the important minor species.[79,80] It conducts research in
pioneer product’s approved NADA) to determine the cooperation with animal producers, drug manufactu-
withdrawal time for their product.[73] Accordingly, rers, FDA’s Center for Veterinary Medicine (CVM),
generic sponsors need only monitor the depletion of The U.S. Department of Agriculture/Cooperative
the unlabeled marker residue in the target tissue to State Research, Education, and Extension Service
establish a withdrawal time. This results in substantial (USDA/CSREES), other government agencies, univer-
time and economic savings. However, there is a risk sities, State Agricultural Experiment Stations, and
that the generic formulation will be associated with a veterinary schools. This group prioritizes and approves
withdrawal time that differs (longer or shorter) than research projects to evaluate safety and effectiveness of
that of the innovator product. For this reason, users needed drugs in minor species. Data from completed
of generic products should carefully read each product research projects are made available as Public Master
label to avoid the presence of violative residues. Files (PMF) that drug manufacturers can reference at
Since veterinary generic drug products must contain no cost to support approval of new animal drugs for
all of the same indications, warnings, cautions, direc- minor species. By 2003, the NRSP-7 was responsible
tions for use, etc. that are associated with the approved for 30 PMFs, involving 13 animal species. An
pioneer product (except as deemed appropriate on the additional 18 research projects were active, involving
basis of the Suitability Petition[77]), a bioequivalence 17 unique animal species and 15 different drugs.
study is generally needed for each species for which Approximately 39% of the active projects involved
the pioneer product is approved on the label, with ruminant species, 11% avian, 33% aquatic, and 17%
the exception of ‘‘minor’’ species. Based upon a survey other veterinary species.
of the 1999 CFR, more than 30% of veterinary pro- The enactment of the Minor Use and Minor Species
ducts approved for oral administration are indicated Animal Health Act of 2004 (MUMS)[81] should help
for use in two or more target animal species. For par- ease the critical shortage of FDA-approved limited-
enteral products, about 45% of the products approved demand animal products, much as the Orphan Drug
in two or more target animal species and approxi- Program has done in human medicine. The MUMS
mately 25% are approved for use in three of more Act created new incentives and procedures to increase
target animal species (note, these estimates exclude the availability of safe and effective drugs for treating
products that are eligible for waivers of in vivo bio- diseases in common domestic species (dogs, cats,
equivalence study requirements). This multiple study horses, cattle, swine, chickens, and turkeys) that occur
(species) requirement increases the risk of failing to infrequently or in a limited geographical location, and
have an approvable application. for treating minor species such as small ruminants, zoo
animals, fish, wildlife, and nontraditional pets. The Act
delineated a process for conditional approval of drugs
Veterinary Pharmaceuticals for Minor for minor uses or minor species; established an index
Uses and Minor Species for legally marketed unapproved drugs for minor
species that are not intended for human or animal
There remains a significant shortage of drugs approved food; authorized research funding for developing these
for use in uncommon species and for treating diseases new drugs; provided a period of exclusivity designated
and conditions that occur infrequently in animals. In for minor use drugs following approval; and author-
aquaculture, the USDA estimates loss of more than ized an office of minor animal species within HHS/
$100 million each year due to fish death resulting from FDA/CVM. With regard to approval requirements,
about 50 different disease conditions.[78] Rare and CVM’s Guidance #61[82] provides an explanation

Veterinary–
valuable zoo animals may die because there is little of the approval requirements and approaches for

Virtual
information on safe and effective treatments. Veteri- demonstrating the safety and effectiveness of these
narians have few options for treating newly emerging applications.
diseases in limited geographic areas. Although there New projects continue to help facilitate drug devel-
is little economic incentive for development of these opment and approved use for minor species and minor
products, approval is needed to define safe and uses. To reduce the expense of drug development in
effective dosages, as well as to determine appropriate aquatic species, efforts are underway to define species
withdrawal times for treated animals intended as food groups (crop grouping) for studies such as TAS, HFS
sources. and effectiveness.[83,84] A searchable aquaculture data-
The National Research Support Project No. 7 base of residues and pharmacokinetic parameters was
(NRSP-7) was created in the United States in 1982 to recently published[85] to make such data more widely
encourage, fund and oversee research on efficacy, available and easily used.
animal safety, HFS and environmental impact of Despite these measures, development of these
drugs needed to preserve the health of agriculturally limited-demand products poses unique difficulties.
 3990 Veterinary Pharmaceuticals: Factors Influencing Their Development and Use

Often, low total numbers of animals make clinical Databank.[91] Accepted indications listed in the USP-
trials difficult to design. Additionally, even the defi- DI do not simply reflect the approved label indication,
nition of minor species differs between countries, but also includes science-based off-label indications.
although efforts to increase drug availability through The credibility of the information is maintained by
harmonization of regulations are underway. For subjecting all USP-DI monographs to an extensive
example, within the United States, minor species are review process by the Expert Panel followed by public
all species other than humans, cattle, horses, swine, review.
chickens, turkeys, dogs, and cats. In the EU, minor The other veterinary drug panel in USP is the
species are those with small total animal numbers, or Expert Committee on Veterinary Drugs. This panel
low economic value for individual animals or their was formed in 2000 to establish public standards pub-
products. But the greatest challenge is the diversity lished in the USP-NF.[92] The monographs published
of species, diseases, management settings, and pharma- in the USP-NF provide pharmacopeial information
ceutical products to be considered. For example, in on definition, packaging storage and other require-
fish, drug pharmacokinetics and systemic ion concen- ment, and a specification that defines the active sub-
trations can markedly change as water temperature stance in the formulation. The monograph provides
increases from 6 to 18 C.[86,87] information on specific tests (such as assay information
Despite these challenges, pharmaceuticals for minor and tests for impurities) the criteria used to define the
uses and minor animal species are sorely needed. active compound, and acceptance criteria. Of the over
Creative approaches and incentives will continue to 4000 monographs in USP-NF, there are 146 veterinary
be needed to bring new products to market while main- drug monographs, with several more in production
taining safety and effectiveness. stages.
Veterinary drug sponsors are encouraged to provide
information and data that can be used to develop new
THE USP monographs. These monographs establish a public
standard to assure practitioners, consumers, regula-
The USP was established in 1820 to provide public tors, and manufacturers that drugs comply with basic
standards for therapeutic products, names, and recipes standards of strength, purity, and quality. The publi-
for their preparation. In 1880, the USP shifted from cation of USP monographs is the result of a collabora-
being simply a recipe book to one that contained medi- tive effort that involves pharmaceutical manufacturers
cation standards for purity, quality, and potency. The and the USP. This process is voluntary, but the USP
USP standard monographs are published in the USP monograph is named as the official compendia of the
and National Formulary (NF), the latter containing United States by the FDA. The advantage to the drug
pharmacopeial information for thousands of drugs sponsor for establishing an official USP drug mono-
and excipients.[88] graph is that as long as there is an official public stan-
In the 1970s, the USP formed the USP-Drug Infor- dard, the less that a company has to rely on developing
mation (USP-DI) Division to develop therapeutic drug private procedures.
information. Advisory panels (Expert Committees) The Expert Committee on Veterinary Drugs is cur-
were created to develop drug monographs that provide rently developing recommendations for in vitro dissolu-
information to health care professionals. A veterinary tion of veterinary solid oral dosage formulations. The
DI panel was established in 1983, first chaired by Dr. current guidelines that exist for human drugs may
Lloyd E. Davis. Today, the USP Veterinary DI Expert not apply to veterinary drugs for a variety of reasons,
Committee consists of 12–15 members (pharmacol- including potential differences in tablet size and
Veterinary–

ogists, clinical pharmacologists, veterinary pharmacists, composition, as well as important differences in the
Virtual

and veterinary practitioners) and USP staff. Their gastrointestinal physiology of humans vs. veterinary
efforts have produced drug monographs for antibio- species.[93] These differences can affect the in vivo rel-
tics,[89] anti-inflammatory drugs,[90] and antiparasitic evance of in vitro dissolution data. Consistent with this
drugs (not yet published). Information contained effort is a recent initiative by the USP Veterinary Drug
within these USP-DI drug monographs include drug Expert Committee to explore the application of in
chemistry, general considerations (such as spectrum vitro–in vivo correlations (IVIVC) to predict bioequi-
of activity for antibiotics), accepted indications, regu- valence for oral drugs administered to dogs using the
latory considerations (including withdrawal times for Biopharmaceutics Classification System (BCS) pre-
label use and off-label use), pharmacokinetics, precau- viously proposed by Amidon et al.[94] and used as the
tions, drug interactions, side effects, veterinary dosing basis for the 2002 guidance published by the FDA
information, and details on dosage forms (including Center for Drug evaluation and Research (CDER) to
brand names). The extralabel withdrawal times are support the biowaiver of certain human oral drug
provided by the Food Animal Residue Avoidance products.[95]
Veterinary Pharmaceuticals: Factors Influencing Their Development and Use 3991

The USP also provides guidelines for drug com- maximum residual limits (MRLs) for veterinary drugs
pounding. The challenges associated with compounded (MRLVD), as well as documents related to the use and
drugs for veterinary use is discussed in Section VIII, F. control veterinary drugs and their residues (e.g., cri-
teria for the validation of analytical methods).
In 1985, the Codex Alimentarius Commission
INTERNATIONAL HARMONIZATION recognized that ‘‘the occurrence and safety of residues
EFFORTS of veterinary drugs in foods of animal origin was of
significance to public health and consumer concern.’’
A range of other organizations support prudent use of Therefore, they established a Codex Committee on
pharmaceutical agents and facilitate the international Residues of Veterinary Drugs in Foods (CCRVDF).
harmonization of veterinary medicine within our global The primary tasks of the CCRVDF include:
environment. Many of theses represent joint ventures
with the WHO, the Food and Agriculture Organization
Determining priority veterinary drugs for the con-
(FAO), and the Joint FAO/WHO Food Standards sideration of residues of veterinary drugs in foods.
Program (Codex).
Recommending maximum levels of such sub-
stances.

Developing codes of good veterinary practice.
Codex Alimentarius Commission
Considering methods of sampling and analysis for
(Codex)[96,97] the determination of veterinary drug residues in
foods.
The volume of world food trade is enormous and is
valued at between US$ 300 billion and 400 billion Proposals of compounds for which MRLs are ela-
(Understanding Codex Alimentarius). borated or proposals for the revision of existing MRLs
During the early 1960s, the FAO Conference and are considered initially by the CCRVDF and then sub-
the WHO launched a joint effort to ‘‘protect the health sequently submitted to the Codex Commission for
of the consumers and ensure fair practices in the food approval as new work. The scientific data assessments
trade.’’ With this in mind, the resulting FAO/WHO for these compounds are undertaken by the Joint
Food Standards Programme works towards the FAO/WHO Expert Committee on Food Additives
following objectives: (JECFA).
JECFA recommendations are used by Codex to ren-

Promote the coordination of all food standards der the final determination of internationally accepted
work undertaken by international governmental MRLs. JECFA provides advice, and CCRVDF makes
and nongovernmental organizations. the risk management decisions. Withdrawal times are

Determine priorities, initiate, and guide the prep- routinely maintained within the domain of each indi-
aration of draft standards with the aid of appropri- vidual national government. The establishment of resi-
ate Codex Committees and organizations. dues standards involves an eight-step procedure,

Finalize standards and, after acceptance by govern- encompassing the initial identification of the need for
ments, and publish them in Codex Alimentarius an MRL, the elaboration of MRL recommendations,
either as regional or world wide standards. debate, comment, and finally acceptance (the Pro-

Amend published standards, after appropriate cedural Manual can be obtained on line at www.
survey in the light of developments. codexalimentarius.net). This eight-step process typically
requires several years from nomination to approval.

Veterinary–
The Codex Alimentarius Commission serves as the

Virtual
seminal global reference point for consumers, food
producers and processors, national food control agen- The Joint FAO/WHO Expert Committee
cies and the international food trade. Codex standards on Food Additives JECFA[101]
and guidelines cover over 200 food commodities and
provided the safe limits for over 3000 contaminants JECFA is an international expert scientific committee
and residues. In 1985, UN Resolution 39/248 adopted that is administered jointly by the FAO and the
guidelines advising governments to support and, as far WHO and evolved from the 1955 FAO/WHO Joint
as possible, adopt standards from the Codex.[98–100] Conference on Food Additives. It has been meeting
The primary product resulting from this program is since 1956, and currently works towards the evaluation
the Codex Alimentarius, which literally means the food of contaminants, naturally occurring toxicants, and
code. This code represents a collection of food stan- residues of veterinary drugs in food. To date, it has
dards and guidance documents published jointly by evaluated more than 1300 food additives, approxi-
the FAO and the WHO. Among these standards are mately 25 contaminants and naturally occurring
 3992 Veterinary Pharmaceuticals: Factors Influencing Their Development and Use

toxicants, and residues of approximately 80 veterinary Recently, the VICH has completed the development
drugs. When a residue has been identified for risk of a Safety Working Group, which is responsible for
assessment, JECFA is responsible for the following evaluating the safety of residues of veterinary drugs
determinations: in human food.

The elaboration of principles for evaluating residue

safety ENHANCING THERAPEUTIC OPTIONS

The establishment of ADIs

The recommendation of MRLs Animal Drug Use Clarification Act of 1994

A determination of the criteria for the appropriate (AMDUCA)
methods of analysis for detecting and/or quantify-
ing residues in food. Prior to 1996, the Federal Food, Drug, and Cosmetic
Act required on-label use of all animal health drug
JECFA serves as a scientific advisory body to FAO, products. This extralabel use restriction precluded
WHO, FAO and WHO member governments and to veterinarians from exercising their clinical judgment
the Codex Alimentarius Commission. Advice to Codex to render therapeutic decisions, be it drug use in an
on veterinary drug residues is normally provided via unapproved animal species, use for an unapproved
the CCRVDF. In the case of veterinary drugs, data indication, or the use of an approved drug in an
on good practices are evaluated and the corresponding approved species at dosage levels higher than that sta-
MRLs in animal tissues, milk, and/or eggs are recom- ted on the label. The use of human drugs for treating
mended. Such MRLs are intended to provide assur- companion animals was also illegal. Thus, veterinar-
ance that when the drug has been used properly, the ians were restricted to a very limited therapeutic
intake of residues of drug present in food is unlikely arsenal and certain kinds of medications (e.g., antican-
to exceed the ADI for that compound. cer drugs) were not legally available. On November 7,
1996, Title 21 of the CFR was amended to add part
530, titled ‘‘Extralabel Drug Use in Animals’’ to the
International Cooperation on Harmonization Food, Drug and Cosmetic Act. AMDUCA allows
of Technical Requirements for Registration veterinarians to prescribe extralabel uses of certain
of Veterinary Products (VICH)[102] approved animal drugs and approved human drugs
to animals under certain conditions. This amendment
The VICH is a trilateral (EU–Japan–USA) program, provided veterinarians with the flexibility necessary
initiated in 1983, whose mission is the harmonizing to meet patient needs.[103]
of technical requirements for veterinary product regis- Post-AMDUCA, veterinarians are permitted to
tration. The objectives of the VICH are along the same prescribe extralabel uses of approved drug.[104] The
lines as those of the ICH which include: key constraints are that any extralabel use must not
result in violative residues in food-producing animals,

Providing a forum for a constructive dialogue the use must be by or on the order of a veterinarian
between regulatory authorities and the veterinary within the context of a veterinarian–client–patient
medicinal products industry on the real and per- relationship, and the use must be in conformance with
ceived differences in the technical requirements for the new regulations, including the exclusion of
product registration in the EU, Japan, and the compounds prohibited from extralabel use in food-
U.S.A., with the expectation that such a process producing animals.[105,106]
Veterinary–

may serve as a catalyst for a wider international

Virtual

harmonization.

Identifying areas where modifications in technical FARAD
requirements or greater mutual acceptance of
research and development procedures could lead Created in 1982, FARAD is a computer-based decision
to a more economical use of human, animal, and support system designed to provide livestock produ-
material resources, without compromising safety. cers, extension specialists, and veterinarians with prac-

Forwading recommendations on practical ways to tical advice on how to avoid drug, pesticide, and
achieve harmonization in technical requirements environmental contaminant residue problems when
affecting registration of veterinary products and to products are used in an extralabel manner.[91] It serves
implement these recommendations in the as the largest repository of animal pharmacokinetic
three regions. Once adopted the VICH recommen- data in the world,[107] maintaining an up-to-date com-
dations should replace corresponding regional puterized compilation of current label information
requirements. (including withdrawal times) on all drugs approved
Veterinary Pharmaceuticals: Factors Influencing Their Development and Use 3993

for use in food animals in the U.S., Canada, Europe, agents while they are at the farm where the animals
and Australia. It also contains the official tolerances are being treated. It promotes the prudent use of anti-
for drugs and pesticides in tissues, eggs and milk, infor- microbial compounds to minimize the selection of
mation on screening tests, and the fate of chemicals in resistant microbial strains. VADS correlates dosages
food animals. with the pharmacokinetics of that compound in the
FARAD is a collative effort between the USDA, target animal species, the susceptibility of the targeted
FDA, and three universities (North Carolina State micro-organism (breakpoints when available), the
University, University of Florida, and University of relationship between drug exposure and antimicrobial
California, Davis). It was authorized by Congress in effect, and clinical effectiveness information when
1998 and is sanctioned to provide these withdrawal available.[110]
time estimates to the USP (Public Law 105–185).[107] VADS is spearheaded by individuals from Iowa
Numerous sources of residue avoidance information State University, Mississippi State University, and Vir-
(including proprietary information provided by drug ginia-Maryland Regional College of Veterinary Medi-
sponsors) are collected and reviewed by residue experts cine. Funding for VADS comes from several veterinary
to insure accuracy and consistency. organizations, two producer organization, and CVM
The focus of FARAD has been and remains as the through an extramural cooperative agreement.[111]
development of recommendations on withdrawal times
associated with extralabel drug use. FARAD offers
emergency response assistance for accidental or delib- CONCLUSIONS
erate chemical exposures to food animals, providing
emergency hotline assistance to State and Federal Narrow profit margins, interspecies differences, and
agencies dealing with chemical contamination in food HFS concerns lead to some distinctly different regulat-
animals.[107] When a field veterinarian contacts FARAD ory and use considerations for veterinary pharmaceuti-
regional access centers, the veterinarian provides infor- cals. Oftentimes, harmonized efforts between industry,
mation on their specific condition of extralabel use governments, and academia are needed to insure
(Table 8). FARAD personnel analyze the information judicious product use while minimizing barriers to
to provide withdrawal interval recommendations. international trade, minimizing public health concerns
FARAD estimates of withholding intervals may (residues and microbial resistance), and insuring
rely on pharmacokinetic data to help define the final environmental safety. It is our hope that this chapter
terminal elimination phase of a drug. In these cases, provided insights into the complexities confronting
it is assumed that the final edible tissue elimination scientists, regulators, and practitioners involved in
phase will be at least as long as that observed in blood. pharmaceuticals and animal health.
Although specific safety factors are not built into these
estimates, the estimated withholding times are
expanded to help ensure that residues drop well below
the established tolerance by the end of the recom- ACKNOWLEDGMENTS
mended period.[108]
The authors would like to express sincere appreciation
to the following individuals for their invaluable com-
The Veterinary Antimicrobial Decision Support ments and suggestions: Dr. Steven Vaughn and Dr.
System (VADS)[109] Joan Gotthardt (general manuscript review); Dr. Mark
Robinson (human food safety); Dr. Margaret Oeller
(veterinary pharmaceuticals for minor uses and minor

Veterinary–
VADS is a web-based database intended to help veteri-

Virtual
narians optimize the use of antimicrobial therapeutic species); and Dr. Richard Ellis (international harmoni-
zation efforts).

Table 8 Examples of extralabel use conditions needing

FARAD input
ARTICLE OF FURTHER INTEREST

The use of drugs for diseases not included in the label
indications
Veterinary Dosage Forms, p. 3941.

The use of extralabel doses

Treatment administration for a duration exceeding
that on the approved product label
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Veterinary–
Virtual
Viral Inactivation Issues in Aseptically Processed Parenterals
J. Hotta
A. Klos
S. Petteway, Jr.
D. Pifat
Bayer Corporation, Research Triangle Park, North Carolina, U.S.A.

INTRODUCTION mixtures that cannot be terminally sterilized and must

be aseptically processed. In addition, all biologics derived
Plasma-derived therapeutic proteins are parenteral bio- from human sources, such as plasma, carry the risk
logics that are purified on an industrial scale. All biologics of viral contamination, as the use of plasma-derived
derived from human sources, such as plasma, carry products has resulted in the transmission of hepatitis
the risk of viral contamination. Thus, in order to market and AIDS. Thus, in order to market a medicinal pro-
a medicinal product derived from human plasma, duct derived from human plasma, manufacturers must
manufacturers must assure the absence of specific viral assure the absence of microbial, as well as specific viral
contamination. Virus validation studies are performed contamination.
to evaluate the capacity of a manufacturing process to Viral contamination of complex biologics can be
remove viral contaminants. Virus clearance across three minimized by strict control of the source material
different terminal inactivation steps, low pH incubation (plasma) and the production process. Controlling the
of immunoglobulins (IgG), pasteurization of albumin, source material involves donor selection, rigorous test-
and freeze dry/dry heat treatment of plasma-derived ing of plasma for virus markers, and inventory hold
products (Factor VIII and Protein G), is discussed in this (Fig. 1). Screening of plasma donors begins with a
article. The data show that, like all other upstream virus detailed health questionnaire to identify high-risk fac-
reduction steps, the methods used for terminal inacti- tors (e.g., family history of Creutzfeldt–Jakob disease)
vation are process and product dependent, and that the and behaviors (e.g., tattoos), followed by a physical
reduction factors for an individual step may be overesti- exam and a background check against the National
mated or underestimated due to inherent limitations or Donor Deferral Registry (NDDR). Source plasma
inadequate designs of viral validation studies. collected by plasmapheresis (i.e., clear liquid remaining
Parenteral drugs are medicinal products that are not after removal and reinfusion of red cells, leukocytes,
ingested but are injected through the skin, directly into a and platelets back to the donor) from qualified donors,
blood vessel, organ, tissue, or lesion. Since the normal rather than recovered plasma (i.e., plasma removed
body defenses against infection are bypassed during from entire blood donations) is used. Qualified donors
parenteral administration, preparation of these drugs are those applicants who have successfully passed two
requires the highest level of contamination control. Con- donor screenings and testing for viral markers within
tamination can be controlled by terminal sterilization or 6 mo. If a donor does not return to donate within
by aseptic processing. Terminal sterilization involves 6 mo, the previous plasma donation is destroyed.
filling final containers with product and sealing them Plasma units from applicants who are disqualified on

Veterinary–
under a high quality, but not necessarily sterile environ- the basis of test results are also destroyed and the

Virtual
ment. The product in its final container is then subjected names of the rejected donors are entered into the
to a sterilization process. During aseptic processing, the NDDR, a national database that lists all donors who
product, container, and closure undergo separate sterili- are excluded from further donations.
zation processes and are then brought together. Since
microorganisms may be inadvertently introduced dur-
ing filling, these operations must be performed under BACKGROUND
stringent conditions to minimize the risk of microbial
contamination (e.g., operating in a clean room environ- Plasma inventory hold refers to the withholding of all
ment, implementing environmental monitoring programs, plasma units from qualified donors for 60 day. If a
and training personnel in aseptic techniques). donor, on a repeat visit, shows evidence of viral infec-
Plasma-derived therapeutic proteins are parenteral tion (e.g., seroconversion) or indicates involvement in
biologics that are purified on an industrial scale. Most potential high-risk activity, the donor’s previous units
biologics are highly complex, heat-sensitive, protein held in inventory can be retrieved and destroyed.
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-120012029
Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved. 3997
 3998 Viral Inactivation Issues in Aseptically Processed Parenterals

biological functions. Because of the complex mixture

of proteins, the inability to apply terminal sterilization
and the need for tight control of source material, the
entire manufacturing process must follow current good
manufacturing practices (cGMP). Biological products
are defined more by their manufacturing processes,
rather than by analytical testing of final product, to
assure the final product is pure, potent, stable, free of
microbial contamination, and carries a low risk of viral
contamination.
Virus validation studies are performed to evaluate
the virus reduction capacity of a manufacturing pro-
cess and a number of regulatory guidance documents
have been issued that describe how to perform and
interpret virus clearance experiments.[1,2] During these
studies, production product intermediates are spiked
with significant amounts of virus, the material is pro-
cessed and virus reduction is measured by comparing
the level of virus infectivity remaining in the final
product-containing fraction with that in the starting
material. Process validation is usually performed using
the same type and size of equipment that will be used
at commercial scale. However, the deliberate contami-
nation of the designated manufacturing equipment and
facility with infectious virus would be unacceptable. As
a result, virus validation studies are conducted using
small-scale replicas and processing conditions rep-
resentative of production scale.
Reduction, across individual steps of a process, is
expressed on a logarithmic scale because virus vali-
Fig. 1 Controlling source material by the screening and dation studies can only assure reduction of the virus
testing of plasma. load by a certain factor. Total reduction of the virus
load to zero cannot be guaranteed. The removal/inacti-
vation of the input virus load to below detection or the
All donations must be tested and found non-reactive reduction of 4 log10, or more, virus is considered signifi-
for syphilis, hepatitis B surface antigen (HBsAg) and cant. Virus reduction factors for process steps that clear
negative for antibodies (Ab) to human immunodefi- the entire input virus load to below detection, are
ciency virus (HIV) types 1 and 2, and hepatitis C virus distinguished from virus reduction factors for steps
(HCV). Many manufacturers are also testing all plasma whose final fractions still contain infectious virus, by
donations that are negative by standard serological test- the symbol ‘‘
.’’ For example, a virus reduction factor
ing, by nucleic acid amplification techniques (NAT) for of
3.7 log10 would indicate that no infectious virus was
the presence of viral genomes such as HCV, HIV-1, detected in the final fraction and that virus reduction
Veterinary–

hepatitis B virus (HBV) and parvovirus B19. Plasma could have been greater than 3.7 log10 if the input virus
Virtual

units that are NAT reactive for HCV, HIV-1 and spike had been greater. A virus reduction factor of
HBV or contain high levels of parvovirus B19 DNA 4.3 log10 means a process step cleared significant levels
are discarded. of virus but residual virus could be detected in the final
The plasma units that meet viral testing standards by product-containing fraction.
NAT are pooled at the production facility and the pools The overall virus reduction capacity of a manufac-
are retested for the presence of selected viral markers turing process is the sum of the individual reduction
(e.g., HBsAg, Ab to HIV-1/2 and HCV, and HCV factors, and should be greater than the potential virus
RNA). Thus, all source material is screened to reduce load in the starting material. At least one step in the
or to confirm the absence of many clinically relevant process should clear significant levels of infectious
viruses and to minimize any potential viral challenge virus so that the overall clearance is not made up of
to the production process. individual small, and possibly negligible, reductions.
During the production process, plasma is frac- Virus reduction steps may be placed upstream or
tionated or separated into fractions with different at the very end of the manufacturing process.
Viral Inactivation Issues in Aseptically Processed Parenterals 3999

Reduction in viral infectivity occurs by inactivation of a wide range of physico-chemical properties, then vali-
virus or by removal of virus particles. During removal dation studies should also include non-specific model
steps, virus is not inactivated but is separated from viruses with many different properties. Inactivation/
the protein of interest using methods such as precipi- removal of these non-specific model viruses provides
tation, chromatography, or filtration. For example, assurance that the manufacturing process is capable of
during an ethanol precipitation step, ethanol is added clearing a diverse range of known and unknown viruses.
to a suspension to precipitate unwanted contaminating The virus reduction studies of the three process steps
proteins and viruses. The ethanol-containing suspen- discussed here were performed with HIV-1, Bovine
sion is then centrifuged so that the contaminants in viral diarrhea virus (BVDV), Pseudorabies virus (PRV),
the precipitated paste fraction can be separated from Reovirus type 3 (Reo), Hepatitis A virus (HAV), and
the product in the effluent fraction. Porcine parvovirus (PPV). HIV-1 was included as a
During inactivation steps, viral infectivity is reduced relevant enveloped virus, while BVDV and PRV were
by treatment with chemicals and/or physical methods. tested as specific model viruses for HCV and HBV,
Remnants of virus particles (e.g., viral nucleic acids) respectively (Table 1). Reo was chosen as a non-specific
may remain in the product-containing fraction but are model non-enveloped virus, HAV was included as a
not infectious. Chemical methods of virus inactivation, relevant virus and PPV was used as a surrogate for
such as treatment with solvent–detergent or acetone, human parvovirus B19. All viruses were propagated
must be placed upstream, since subsequent steps are using standard cell culture conditions.[3,4] The appro-
needed to remove or reduce the levels of the toxic priate cell lines were infected, at a low multiplicity of
chemicals. Terminal inactivation is often achieved using infection, and incubated until 4þ cytopathic effects were
physical methods, such as heat and low pH, because observed. The infected cells were frozen and thawed three
these methods leave no chemical residues. After treat- times to release virus, centrifuged at low speed to remove
ment, the final products are delivered to patients, so cell debris and the clarified supernatants were removed
aseptic processing conditions must be maintained for use as virus spikes.
throughout terminal inactivation steps and the param-
eters for virus inactivation must be balanced with the
conditions to preserve product quality and yield. Terminal Inactivation Step: Low pH Final
The focus of this article is to discuss virus reduction Container Incubation of IgG
issues that arose during studies of three different
terminal inactivation steps: low pH incubation of immu- Although the partitioning of viruses during ethanol
noglobulins (IgG), pasteurization of albumin, and freeze fractionation and the presence of neutralizing anti-
dry/dry heat treatment of plasma-derived products bodies are thought to be the main factors involved in
(Factor VIII and Protein G). The data show that, like establishing the safety of immunoglobulins,[5] trans-
all other upstream virus reduction steps, the methods mission of HCV by intravenous immunoglobulin
used for terminal inactivation are process and product (IVIg) has been reported.[6] Many IgG preparations
dependent, and the reduction factors for an individual are formulated at low pH and held for extended peri-
step may be overestimated or underestimated due ods. The low pH hold step was originally designed to
to inherent limitations or inadequate designs of viral stabilize IgG, reduce anticomplement activity, and allow
validation studies. for intravenous administration.[7,8] Virus validation
studies showed incubation at low pH also inactivated
significant levels of enveloped viruses.
SELECTION OF VIRUSES FOR VIRUS The three human IgG products discussed here were

Veterinary–
VALIDATION STUDIES purified from Fraction II þ III paste by slightly dif-

Virtual
ferent processing methods but all three shared the same
A major issue in performing virus validation studies is final formulation in 0.2 M glycine, pH 4.25. All three
determining which viruses should be used. The Com- were 10% protein solutions, of which 98% was IgG,
mittee for Proprietary Medicinal Products (CPMP) and their monomer contents were greater than 90%.
has issued guidelines on the selection of viruses to evalu- At production scale, IgG final bulks (pH 4.25) are typi-
ate in validation studies.[1] Processes must be validated cally sterile filtered, aseptically filled into sterile final
for their capacity to inactivate/remove relevant viruses, containers and incubated at 20–27 C for 21 days–
or viruses that are known to contaminate plasma or 28 days.[9] For the virus studies, however, IgG final
other materials in the production process. If relevant bulk material was spiked with virus and adjusted to
viruses cannot be easily propagated in cell culture or pH 4.3 or 4.5 before incubating at 5, 20, or 23 C, for
assayed, then validation studies should include specific up to 28 days. Hanks’ Balance Salt Solution (HBSS) or
model viruses with characteristics similar to relevant IgG, adjusted to pH 7, were also spiked with virus
viruses. If relevant viruses do not represent viruses with and included as positive controls. Aliquots for virus
 4000 Viral Inactivation Issues in Aseptically Processed Parenterals

Table 1 Test viruses

Enveloped viruses

Human immuno-deficiency Bovine viral diarrhea Pseudorabies virus

Characteristic virus (HIV) virus (BVDV) (PRV)
Relevant or model for HIV-1, HIV-2 HCV CMV, EBV, HSV, HBV
Strain (III B) Kentucky-22 (BRFF) PRV dl tk (ATCC VR-2074),
Becker (Dupont/Merck)
Nucleic acid RNA RNA DNA
Enveloped Yes Yes Yes
Size (nm) 80–130 80–100 120–220
Resistance to Low Medium Medium
physico-chemical agents
Non-enveloped viruses

Reovirus type 3 (Reo) Hepatitis A virus (HAV) Porcine parvo virus (PPV)
Relevant or model for Non-enveloped virus HAV Human parvovirus B19
Strain Abney (ATCC VR 232) HM175 18f (ATCC) NADL-2 (ATCC VR 742)
Nucleic acid RNA RNA RNA
Enveloped No No No
Size (nm) 60–80 27–32 18–26
Resistance to High Very high Very high
physico-chemical agents

titration were removed on day 0 and at various times 4.2 log10 at pH 4.3 but at pH 4.5, BVDV reduction
during incubation. was only 2.6 log10.
The extent and kinetics of reduction were dependent BVDV reduction was also a function of the IgG test
on the test viruses and the exact processing conditions solution. After 21 days in IgG solution 3, pH 4.3, 5 C, no
(e.g., pH, time, and temperature). Virus inactivation virus could be detected but approximately 4 log10 BVDV
was greater with lower pH, higher temperatures and was still present in IgG solution 1 under the same con-
longer incubation times and the kinetics of HIV-1 ditions. For IgG solution 1, significant BVDV reduction
and PRV inactivation were more rapid than BVDV. (4.2 log10) was observed at pH 4.3 at 20 C and 23 C but
HIV-1 and PRV were always inactivated to below at pH 4.5, less than 4 log10 inactivation was achieved.
detection after incubating for 14 days at low pH, In contrast, significant BVDV reduction (greater than
20–23 C but the kinetics of inactivation were slightly 4 log10) was observed in IgG solution 3 at both pH 4.3
different in different IgG solutions (Fig. 2 and and 4.5, regardless of temperature.
Table 2). The entire input load of HIV-1 or PRV was The results are consistent with other studies in dem-
inactivated sooner in IgG solution 1 than in IgG solu- onstrating the effectiveness of low pH incubation to
Veterinary–

tions 2 or 3. HIV-1 was below detection after 21 days in inactivate enveloped viruses. After incubating IVIg
Virtual

IgG solution 1, regardless of pH and temperature, but (6% protein, 8% sucrose) at pH 4.4 for one week at
significant levels of infectious virus were still present 35 C, or for four weeks at 23 C,
6 log10 Vesicular
after 21 days in IgG solution 2, pH 7 or 5 C. These Stomatitis Virus (VSV) was inactivated.[10] IVIg (8%
discrepancies could be due to minor differences in protein, 16% maltose) incubated at pH 4 (in the pres-
pH (pH 4.3 vs. pH 4.5), temperature (23 C vs. 20 C), ence of pepsin) for 22 hr at 35 C inactivated mumps,
or virus preparation. Semliki Forest Virus (SFV), Herpes simplex virus
To control these experimental differences, BVDV (HSV), and vaccinia virus but had no effect on the
inactivation was monitored, in parallel, during low non-enveloped poliovirus type 2.[11] IVIg (7–8% pro-
pH incubation, in IgG solutions 1 and 3, using the tein, glucose-containing buffer) at pH 4.25 (in the
same stock of virus (Fig. 3 and Table 2). The data show presence of pepsin), 37 C, for 30 hr inactivated HIV,
minor changes in pH and temperature resulted in PRV, and BVDV to below detection.[12]
major differences in virus reduction. For example, Immunoglobulins are biologics that differ in many
BVDV reduction in IgG solution 1, 20 C, was respects from classically synthesized drugs as there
Viral Inactivation Issues in Aseptically Processed Parenterals 4001

Fig. 2 Kinetics of virus inactivation during low pH incubation in IgG solutions. Virus was spiked into IgG solutions, before
adjusting to pH 7, 4.5, or 4.3, and the solutions were incubated at 5, 20, or 23 C. HBSS was also spiked as a positive control.
Aliquots for virus titration were removed immediately after spiking (t ¼ 0 day) and at various times during incubation. IgG sol-
ution 1: HIV-1 (A) or PRV (C), IgG solution 2 or 3: HIV-1 (B) or PRV (D), dashed line, no symbol ¼ virus detection limit.

Veterinary–
are no chemically defined formulas for the many differ- pH (e.g., pH 4.0). Thus, data which demonstrate the

Virtual
ent classes of IgG molecules, each with their own virus safety of a product should not be transferred to
unique antigen recognition capability. Differences in other products, even though the two products may
upstream processing may yield product with slightly appear to be biochemically similar, and each process
different protein profiles and these changes could should be carefully validated.
impact virus reduction. For example, in the studies pre-
sented here, IgG solutions 1 and 3 were 10% protein
solutions with the same formulation but the kinetics Terminal Inactivation Step: Pasteurization
and extent of virus inactivation were different in the of Albumin
two products. After incubating at pH 4.5, 20 C in
IgG solution 3, BVDV inactivation was 4.1 log10 but Pasteurization is the heating of aqueous, stabilized
only 2.6 log10 in IgG solution 1. Significant reduction protein solutions. Pasteurization has been used for
of BVDV in IgG solution 1 will most likely require bulk preparations of Factor VIII, antithrombin III,
incubation at higher temperatures (e.g., 25 C) or lower and alpha1-antitrypsin but wet heat treatment of these
 4002 Viral Inactivation Issues in Aseptically Processed Parenterals

Table 2 Summary of virus reduction during incubation at low pH

Log10 virus reduction

HIV-1a PRVb BVDVc

Test solution pH 5 C 20/23 C 5 C 20/23 C 5 C 20 C 23 C

HBSS 7
6.6 & 1.2 1.0 1.6 & &
IgG solution 1 7 &
6.5 &
4.3 & & <1.0
4.5 & & & & & 2.6 3.6
4.3
6.5
6.5
3.9
4.3 1.8 4.2 4.2
HBSS 7 na & <1.0 & na & &
IgG solution 2 or 3 7 <1.0 <1.0 <1.0 1.6 & & 1.2
4.5 2.5
4.5
6.3
6.1 & 4.1
4.1
4.3 & & & &
4.5
4.5
4.4
na ¼ not applicable; gray boxes ¼ not done.
a
HIV-1 studies with IgG solutions 1 and 2 were conducted at different times with different virus stocks and slightly different
temperatures. Solution 1 study was 28 days, the incubation temperatures were 5 and 23 C, and titers were calculated as
SFU/ml. Solution 2 study was 21 days, the incubation temperatures were 5 and 20 C and titers were calculated as TCID50/ml.
b
PRV studies with IgG solutions 1 and 3 were conducted at different times, using different virus stocks and slightly different
temperatures. Solution 1 study was 14 days and the incubation temperatures were 5 and 23 C, while solution 3 study was 9 days
and the incubation temperatures were 5 and 20 C.
c
BVDV studies with IgG solutions 1 and 3 were conducted at the same time, using the same virus stock and lasted 28 days.

proteins requires the addition of stabilizers that must be sequestered in a ‘‘sampling neck’’ of a bulk processing
removed by a downstream process step.[13] Removal of container and had not been adequately heated. In con-
stabilizers and denatured proteins from pasteurized trast, final containers of albumin, made by the same
materials extends processing times and may involve sig- manufacturer, were completely submerged in a water-
nificant losses in yield.[14] The development of antibodies bath before heating and did not transmit HBV.[18,19]
to FVIII is a complication of treatment and pasteuriza- The data discussed here are for PlasbuminÕ-25,
tion of FVIII concentrates raised concerns regarding the which is purified from Fraction V paste and consists
possible induction of FVIII neoantibodies.[15] of 23.5%–26.5% protein, of which no less than 96.5%
Pasteurizing at 60 C, for 10 hr, in final container, is is albumin. The preparation is stabilized with 0.02 M
the pharmacopoeial method to inactivate viruses in sodium caprylate and 0.02 M acetyltryptophan and
albumin preparations. Sodium caprylate and acetyl- contains 145 mEq/L sodium.[20] For the virus clear-
tryptophan are often added as stabilizers, which bind ance experiments, virus was spiked into albumin, pH
to albumin to prevent denaturation or aggregration 6.4–7.4, and the solution was heated at 60 C for
during heating. So far, only albumin preparations can 10 hr. Aliquots for virus titration were removed as
withstand pasteurization in the final container because soon as virus was added (preheat), when the tempera-
the quantity of caprylate and acetyl tryptophan added ture of the solution reached 60 C (0 hr) and at various
are compatible for intravenous administration.[16] times during the pasteurization cycle. Unheated albu-
Pasteurization is an effective inactivation step for min and HBSS were also spiked to the same dilution
Veterinary–

both enveloped and non-enveloped viruses, as other with virus and tested as positive controls.
Virtual

investigators have shown pasteurization of 20% albu- Enveloped virus inactivation was biphasic, as
min resulted in
5 log10 HIV-2,
7.5 log10 MuLV, HIV-1, PRV, and BVDV were quickly inactivated

3.5 log10 PRV, 4.2 log10 Sindbis,
5.5 log10 Polio, within the first 2 hr of pasteurization. Although inacti-
and 6.6 log10 SV40 reduction.[17] The strongest evidence vation was much slower after 2 hr, all enveloped virus
for safety is in the clinical setting where historical data infectivity was below detection after 5 hr (Fig. 4).
show that albumin (and plasma protein fraction) solu- Although not completely inactivated, the levels of non-
tions have not transmitted viral disease since pasteuri- enveloped viruses, Reo and HAV (strain HM175/18f),
zation in final container was introduced.[18] decreased significantly, resulting in 5.6 and 4.4 log10
Evidence for the increased assurance of safety from reduction, respectively (Fig. 4 and Table 3). PPV was
heating in final container rather than heating in the most resistant to pasteurization, as less than 2 log10
bulk comes from the 1973 outbreak of HBV that was virus reduction was observed (Fig. 4 and Table 3).
associated with plasma protein fraction (PPF). Investi- Additional experiments were performed to examine
gations revealed that a small amount of PPF had the robustness of pasteurization to inactivate the most
Viral Inactivation Issues in Aseptically Processed Parenterals 4003

Fig. 3 Kinetics of BVDV inactivation during low pH incubation in IgG solution 1 or 3 at 23 C (A) or 20 C (B). The methods
were as described in Fig. 2.

physico-chemically resistant viruses, HAV and PPV. acetyl tryptophan) are also five times higher. Thus,
For these experiments, one parameter (e.g., pH, tem- the discrepancy in HAV reduction in 25% and 5% albu-
perature, or protein concentration) was deliberately min may be related to the concentrations of stabilizers
set at the extreme limit for processing or set outside (or virus destabilizers) in the solutions.
normal production processing ranges. All other param- All viruses used in clearance studies are actually
eters were maintained at the production setpoints model viruses since they have been adapted to grow
(Fig. 5 and Table 3). The data showed that PPV in cell culture and laboratory adapted virus strains
reduction did not change significantly under any of may be slightly different from clinical isolates. Anti-
the conditions tested, while HAV reduction was affec- genic and genetic variations have been reported for
laboratory adapted strains of HAV.[21] Two different
Veterinary–
ted by changes in temperature and protein concen-

Virtual
tration. When heated below or at the lower limit for clones of HAV strain HM175, 18f and 24a, were pas-
temperature, HAV reduction was less than 4 log10. In teurized in 25% and 5% albumin and their kinetics of
25% and 26.5% protein, greater than 4 log10 HAV inactivation were compared. The results show HAV
reduction was achieved but at 5% and 7.5% albumin, HM175/18f was much more resistant to pasteurization
HAV reduction was approximately 3 log10. Lower than HAV HM175/24a (Fig. 6, Table 4). The titers of
reduction at lower protein concentrations was unexpec- HAV HM175/18 were slightly higher than that in the
ted since higher concentrations of protein are expected previous experiment but the extent and kinetics of
to provide virus with better protection from physico- inactivation were similar. HAV HM175/18f inacti-
chemical treatments than lower concentrations. The vation in 25% albumin was 4.5 log10 but was 3.1 log10
ratio of stabilizer to protein is the same for both 5% in 5% albumin. Inactivation of HAV HM175/24a
and 25% albumin. However, since 25% albumin con- was not significantly affected by protein concentration,
tains five times more protein than 5% albumin, the as 4.2 log10 reduction was observed during pasteuriza-
concentrations of stabilizers (sodium caprylate and tion in both 25% and 5% albumin.
 4004 Viral Inactivation Issues in Aseptically Processed Parenterals

Fig. 4 Kinetics of virus inactivation during pasteurization of 25% albumin. (A) HIV-1, (B) PRV, (C) BVDV, (D) Reo, (E) PPV,
and (F) HAV. Virus was spiked into albumin and an aliquot was removed for immediate titration (t ¼ Preheat). Timing of the
pasteurization cycle started when the temperature reached 60 C (t ¼ 0 hr). Unheated albumin and Hanks’ Balanced Salt Sol-
ution (HBSS) were also spiked and incubated at 5 C, as positive controls. Aliquots for virus titration were removed at various
times during pasteurization (closed circles ¼ HBSS, 5 C, closed triangles ¼ 25% albumin, 5 C, closed squares ¼ 25% albu-
min, 60 C, dashed line, no symbol ¼ virus detection limit).

Laboratory adapted strains of HAV are often may be a parvovirus-specific trait, which is not due
associated with cellular membranes. Propagation of to virus association with cellular membranes.
HAV involves freezing and thawing of infected cells In summary, pasteurization is effective for inactivation
to release virus. During its release from disrupted of many enveloped and non-enveloped viruses but their
cells, virus may become entangled or associated with kinetics of inactivation are different. In the experiments
cellular membranes, which may shield virus from neutra- with 25% albumin, enveloped viruses were below detec-
lization by antibody.[22] To determine if cell associated
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tion after heating for 5 hr, leaving a wide margin of safety.

Virtual

lipids protect virus from heat denaturation, HAV In contrast, non-enveloped virus reduction was slower
HM175/18f was extracted with chloroform and inacti- and required the entire 10 hr of heating. Temperature,
vation of the extracted virus and untreated virus, during time, protein concentration, and possibly stabilizers were
albumin pasteurization, was compared. The results show critical parameters for virus inactivation.
inactivation of chloroform-extracted virus was signifi- The data also demonstrated that the subtle differences
cantly faster than untreated virus (Fig. 7). in viruses should not be underestimated. The different
PPV was also extracted with chloroform, but HAV strains/clones and the method of virus prepa-
chloroform extraction had little effect on PPV inacti- ration had a significant impact on virus clearance. Virus
vation during albumin pasteurization (Fig. 7). These stocks used in virus validation studies are produced in
results are comparable to previous reports of only cell culture and the behavior of tissue culture derived
1–2 log10 Minute Virus of Mice, another parvovirus, viruses may be different from that of native viruses. Lab-
inactivation after 6 hr of pasteurization in 1% albu- oratory adapted strains of viruses may also have unpre-
min.[23] Thus, heat stability during pasteurization dicted properties, such as association with lipids, which
Viral Inactivation Issues in Aseptically Processed Parenterals 4005

Table 3 Summary of virus reduction during pasteurization of 25% albumin

Log10 virus reduction

Test parameter HIV-1 PRV BVDV Reo PPV HAV

pH 6.4 & & & & 2.3
4.6
6.9/7.0
5.9
4.8
5.2 5.6 1.6 4.4
7.4/7.5 & & & & 1.7 4.5
Temperature ( C) 58 & & & & 1.6 3.2
59 & & & & 1.9 3.3
59.5 & & & & & 3.9
60
5.9
4.8
5.2 5.6 1.6 4.4
60.5 & & & & &
4.4
Protein concentration (%) 5.0 & & & & 1.7 3.3
7.5 & & & & 1.9 3.5
25.0
5.9
4.8
5.2 5.6 1.6 4.4
26.5 & & & & 1.5 4.6
Gray boxes ¼ not tested.

may affect their properties. Regardless of the panel of products with liquid formulations for some of the
test viruses used in clearance studies, all virus stocks used following reasons:
should be strictly controlled and produced from quali-
fied and traceable cell lines and virus seed banks. 1. Assuring specified fill volumes is difficult because
Repeated passage of the original stock virus is not many formulations consist of small amounts of
recommended because mutations and deviations from the active agent and large amounts of stabilizers.
the original reference material can result. Unlike a powder or liquid fill, a low fill is not
readily apparent after freeze drying.
2. The number of variables to ensure aseptic pro-
Terminal Inactivation Step: Dry Heat cessing is high. Before aseptic fill, different parts
Treatment of Freeze Dried Product of the final product must be sterilized by different
methods, such as dry heat to sterilize and depyro-
Proteins undergo chemical (e.g., deamidation or oxi- genate glass containers, autoclaving and drying
dation) and/or physical degradation (e.g., aggregation to sterilize and remove moisture from rubber
and precipitation) in aqueous solutions. Many of these stoppers, and sterile filtration for the liquid for-
destabilizing reactions are minimized when water is mulated product. After aseptic fill, the stoppers
removed and proteins remain in a dried solid state. are placed on top of the vials and the partially
This process of removing water from the product is stoppered vials are transported, loaded into a
called freeze drying or lyophilization, and is an effec- freeze dryer, and then freeze dried. During freeze
tive way to improve the stability and extend the drying, the sterilized material in the vials in the
shelf-life of labile proteins. During the freeze drying freeze dryer will be exposed to the chamber’s
step, final containers are aseptically filled, the product nitrogen or air, so bacterial retentive filters must

Veterinary–
is frozen and then the frozen solid is dried by the sub- be used in all vacuum/air lines to the chamber.

Virtual
limation of ice under vacuum. Thus, the risk of contamination exists through-
Many freeze dried coagulation concentrates are out the process until the vials are actually stop-
heated to reduce the risk of virus transmission. During pered at the end of the freeze drying cycle.
terminal dry-heat treatment, freeze dried FVIII concen- 3. Equipment reuse also carries a risk of contami-
trate in final container is heated for periods ranging nation, so cleaning procedures for freeze dryers
from several minutes to several days at temperatures should be validated for their effectiveness to
ranging from 60 to 100 C.[4,24–26] This method was orig- inactivate virus.
inally developed to inactivate hepatitis viruses[27] but 4. The technology for freeze drying/dry heat treat-
was later recommended by the National Hemophilia ment is complex and there are many process para-
Foundation[28] and the Centers for Disease Control[29] meters to control (e.g., temperatures of the shelf,
to reduce the transmission of HIV. product and condenser; freezing and drying rates;
Process validation associated with a freeze dried/ pressures of the chamber and condenser; and
dry heated product is more challenging than for temperature and distribution of heat in the oven).
 4006 Viral Inactivation Issues in Aseptically Processed Parenterals

Fig. 5 Effect of (A) pH, (B) temperature, and (C) protein concentration on PPV and HAV inactivation during pasteurization of
Veterinary–

25% albumin. Methods were as described in Fig. 4, except different pH, temperature, or protein concentrations were tested.
Virtual

A process step with many variables to control is HIV (
3 log10 HIV-1 reduction), but was less
equally difficult to scale down for bench scale studies. effective for HBV-like viruses (>1 log10
However, results from animal studies and human clini- PRV reduction, 2.4 log10 VSV reduction) and
cal trials have been consistent with the ‘‘in vitro’’ for HCV-like viruses (2.9 log10 Sindbis
model virus clearance studies of the terminal freeze reduction).[30] Studies in animals and humans
dry/dry heat treatment. confirmed the model virus results. Chimpan-
zees inoculated with Hemofil T, that had been
1. Virus clearance studies of Factor VIII (Hemofil T) spiked with HBV (30,000 chimpanzee infec-
indicated the freeze dry/dry heat treatment tious doses) and freeze dried/dry heated, later
(60 C, 72 hr) step was effective in inactivating developed hepatitis B.[27] During clinical trials,
Viral Inactivation Issues in Aseptically Processed Parenterals 4007

Fig. 7 Effect of chloroform extraction on virus inactivation

during pasteurization in albumin. For these experiments, the
virus spike was divided into half. One-half was extracted with
an equal volume of chloroform while the other-half was pre-
Fig. 6 Kinetics of HAV strain HM175/18f or HM175/24a pared as usual. The untreated and chloroform extracted virus
inactivation during pasteurization in 25% or 5% albumin. were then used as virus spike in pasteurization experiments as
The methods were as described in Fig. 4. described in Fig. 4.

none of the patients treated with heated Hemo- KoateÕ-DVI is a freeze dried concentrate of human
fil T became HIV-1 positive[31] but 11 out of 13 plasma-derived Factor VIII (FVIII) that is stabilized
patients developed non-A, non-B hepatitis.[32] with albumin (10 mg/ml) and formulated in a buffer
2. Virus clearance studies of a solvent/detergent containing 0.05 M glycine, 3 mM calcium, 0.06 M histi-
treated, freeze dried/dry heated (100 C, dine.[34] After freeze drying, it is terminally heated in
30 min) Factor VIII concentrate (Emoclot), final container for 72 hr at 80 C. Water molecules
demonstrated significant inactivation of HAV normally help to maintain proteins in their native state
and poliovirus 1 by the terminal freeze dry/dry but, in freeze dried material, water molecules may
heat step.[24] No model parvovirus was included contribute to degradative reactions. The following stu-
as a test virus in these in vitro studies. During dies were performed to demonstrate that controlling

Veterinary–
clinical trials, 13 previously untreated patients residual moisture levels in freeze dried/dry heated

Virtual
were given Emoclot and none of them became cakes may be a method for achieving viral inactivation
anti-HAV positive. In contrast, B19 seroconver- in the Koate-DVI process.[4]
sion and viremia was observed in 5 of 11 suscep- Vials of virus-spiked FVIII were freeze dried, sealed
tible patients.[33] with stoppers containing different amounts of moisture,
and then heated at 80 C for various times. Moisture
in the virus-spiked samples was monitored by Near
Table 4 HAV reduction during pasteurization in albumin InfraRed (NIR) spectrometry, whose values had been
correlated to loss on drying (LOD) moisture values.[37]
Log10 HAV reduction
After converting the NIR values to LOD moisture
Test solution HM 175/18f HM175/24a levels, the freeze dried/dry heated materials were recon-
25% albumin 4.5 4.2 stituted and titrated for residual virus.
As shown in Figs. 8 and 9, the moisture content in
5% albumin 3.1 4.2
freeze dried material was not constant after freeze drying.
 4008 Viral Inactivation Issues in Aseptically Processed Parenterals

significant levels of enveloped and non-enveloped

viruses if a minimum threshold level of moisture
(0.8%) is attained. Increased moisture levels during
dry heat treatment also destroys FVIII activity and
at 1.6% moisture, the upper limit for moisture at this
step, specific activity decreases an average of 11%.[38]
To compensate for the loss in potency, the bulk is
made more concentrated to assure FVIII activity at
9 IU22 IU activity per mg protein.
Other investigators have also shown parvovirus
inactivation by the terminal freeze dry/dry heat
(80 C, 72 hr) step. Canine parvovirus, spiked into
FVIII concentrate, ReplenateÕ, was inactivated by
3.0 log10, while 4.7 log10 bovine parvovirus spiked into
FVIII concentrate, LiberateÕ,[39] was inactivated after
heating lyophilized product for 72 hr, 80 C. Unfortu-
nately, in these studies, product loss after treatment
was as high as 30%.
Although viruses from different animal species
(e.g., porcine, canine, and bovine) were used to test the
terminal freeze dry/dry heat treatment of the KoateÕ-
Fig. 8 Kinetics of enveloped virus inactivation during 80 C DVI, ReplenateÕ, and LiberateÕ processes, the
heat treatment of freeze dried Factor VIII (FVIII) concen- differences in parvovirus reduction and product recov-
trate. (A) HIV-1, (B) BVDV, (C) PRV (>0.8% moisture) ery were probably due to the different formulations
and (D) PRV (<0.8% moisture). Virus was spiked into FVIII and freeze drying cycles of all three FVIII products.
concentrate and a sample was immediately removed for virus For freeze dried products, formulation and the freeze
titration (pre-FD). Vials were aseptically filled with virus- drying process are interrelated. Without the appropri-
spiked FVIII concentrate and then freeze dried. After freeze ate buffers and stabilizers or optimal temperatures
drying, the vials were closed, using stoppers containing differ- and cycle times, many protein preparations would be
ent levels of moisture, and sealed before heating at 80 C for
denatured by the physical stresses associated with the
up to 72 hr. Near infrared (NIR) readings were taken on vials
freeze dry/dry heat treatment.
after freeze drying (0 hr) and at various times after heating at
80 C (24 hr, 48 hr, or 72 hr). After converting the NIR read- The impact of formulation on virus reduction and
ings to loss on drying (LOD) moisture values, the product in product recovery (e.g., potency) was evaluated during
each vial was reconstituted and assayed for residual virus terminal freeze dry/dry heat treatment studies with
(gray boxes ¼ log10 virus titer, closed circles ¼ % moisture, an unlicensed Fraction I derived product, that will be
and dashed line ¼ virus detection limit). designated ‘‘Protein G.’’ As shown in Fig. 10, the
optimum formulation, that achieved
4 log10 PPV
inactivation and 80% product recovery, was one that
Autoclaved rubber stoppers hold measurable amounts contained 2% albumin, no NaCl, and 0.3% moisture
of water that are transferred to the freeze dried pro- by the Karl Fischer coulometric method. In contrast,
duct[35,36] and heating at 80 C accelerates this transfer. freeze dry/dry heat treatment of product formulated
Virus reduction data are presented in Figs. 8 and 9 with no albumin, 150 mM NaCl and low moisture
Veterinary–

and Table 5. Little or no HIV, BVDV and PPV resulted in approximately 3 log10 PPV reduction and
Virtual

reduction occurred during the freeze drying process, 35% product recovery. Thus, minor changes in formu-
but PRV, Reo, and HAV titers, before (Pre-FD) and lations such as the addition of 2% albumin may impact
after freeze drying (0 hr), decreased 1–2 log10. Regardless virus reduction and product recovery during a freeze
of moisture content, HIV, and BVDV were inactivated dry/dry heat treatment.
to below detection after heating 80 C, 24 hr, and Reo In summary, the large number and complexity of
was completely inactivated after 48 hr. Inactivation of the variables to control when implementing a terminal
HAV, PRV, and PPV was dependent on the moisture freeze dry/dry heat treatment makes validation very
levels in the cake. When the moisture in the system was difficult and increases the probability for error. Since
0.8% or greater, virus inactivation was approximately the reliability of the results from virus clearance studies
4 log10. However, when less than 0.8% moisture was is dependent on the appropriateness of the models used
present, virus reduction after heating was 2.5 log10. in the studies, it is essential that the freeze dry/dry
These data show the freeze dry/dry heat treatment heated material and the experimental conditions used
of the KoateÕ-DVI process can effectively inactivate at small scale be representative of production scale.
Viral Inactivation Issues in Aseptically Processed Parenterals 4009

Fig. 9 Kinetics of non-enveloped virus inactivation during 80 C heat treatment of freeze dried Factor VIII (FVIII) concentrate
(A) in the presence of high cake moisture (>0.8% moisture) or (B) in the presence of low cake moisture (<0.8% moisture). Methods
were as described in Fig. 8 (gray boxes ¼ log10 virus titer, closed circles ¼ % moisture, and dashed line ¼ virus detection limit).

During the studies, temperature and pressure must rubber stoppers should be examined, as they influence
be controlled during all stages of the freeze drying the rate and extent of viral inactivation and the recov-
cycle and uniform temperatures must be distributed ery of biological activity.
throughout the oven used for the dry heat step. Process Perhaps more difficult to duplicate or control dur-
parameters such as the amount of moisture retained by ing small scale virus clearance studies, are the precise

Table 5 Summary of virus reduction during 80 C heat treatment of freeze dried AHF
HIV-1 BVDV PRV

Log10 virus Log10 virus Log10 virus

Veterinary–
Enveloped viruses reductiona Moistureb reduction Moisture reduction Moisture

Virtual
Low moisture
5.3 1.3 (24 hr)
5.7 1.1 (24 hr) 2.1 0.7 (72 hr)
High moisture & & & & 4.0 2.0 (72 hr)
HAV PPV Reo

Log10 virus Log10 virus Log10 virus

Non-enveloped viruses reduction Moisture reduction Moisture reduction Moisture

Low moisture 0.1 0.7 (72 hr) 2.5 0.5 (72 hr)
6.2 0.4 (72 hr)
High moisture
4.5 1.3 (72 hr) 3.7 1.3 (72 hr)
5.5 1.3 (72 hr)
Gray boxes ¼ not tested and methods were as described in Fig. 8.
a
Log10 virus reduction ¼ Log10 total virusPre-FDLog10 total viruspost-80 C heat.
b
Freeze dried product was measured by NIR spectrometry. The NIR values were then converted to LOD moisture values using a NIR/LOD
calibration curve.
 4010 Viral Inactivation Issues in Aseptically Processed Parenterals

Fig. 10 Effect of formulation on virus reduction and product recovery. Protein G was formulated in buffer containing albumin
(0% or 2%) and NaCl (0 mM or 150 mM). Virus was added and a sample was immediately removed for virus titration (Pre-FD).
The virus-spiked material was aseptically filled into vials, freeze dried, and then heated at 80 C, 72 hr. Mock-spiked Protein G
was processed like the virus-spiked samples but was used to measure product recovery (potency).

composition and the molecular structure of freeze Screening and selection of the source plasma will
dried materials. The formulation studies with Protein only avoid contamination by known pathogens. The
G indicated virus reduction and product recovery were protein purification steps and specific virus reduction
influenced by the presence of albumin at a prefreeze methods used in production processes, however, will
drying volumetric concentration of 2%. All proteins inactivate and/or remove both known and unknown
have unique physico-chemical properties and stabiliza- viruses. Terminal virus inactivation treatments are
tion requirements, so formulation and freeze drying applied to product in final container and must balance
cycle parameters must be customized for each new virus inactivation with any modifications to protein
protein drug. For terminal viral inactivation steps, it immunogenicity, activity, and yield. While many
is equally important to develop a formulation and upstream virus inactivation steps rely on chemical
freeze dry cycle that will not stabilize virus. methods that involve the addition and subsequent
removal of toxic agents (e.g., solvent/detergent), physi-
cal methods for virus inactivation, such as pH and
CONCLUSIONS heat, are used for terminal steps.
Virus validation studies assess the virus clearance
Veterinary–

The direct testing for potential viral contamination in a capacity of a process and the evaluation and interpret-
Virtual

finished product is not considered sensitive or accurate ation of virus clearance data from terminal inactivation
enough to assure the absence of infectious virus because and upstream process steps are similar. Limitations in
many of the viruses known to contaminate plasma the design and execution of virus validation studies
(e.g., HCV, HBV, and parvovirus B19) cannot be easily may lead to an incorrect estimate of the ability of a
grown or assayed in cell culture. Testing for the presence process to inactivate/remove virus infectivity. The three
of viral nucleic acids in final product is sensitive but can- terminal inactivation treatments discussed here illus-
not distinguish infectious particles from non-infectious trate the importance of rigorously controlling virus
particles. Thus, assuring the safety of plasma-derived validation studies.
products with respect to virus transmission is an indirect
combinatorial approach: screening and selection of the 1. During the low pH incubation step of IgG,
source material, validation of the production processes minor changes in pH and temperature, as
for inactivation/removal of virus infectivity, and pro- well as in upstream processing steps, signifi-
cess control by strict adherence to cGMP. cantly impacted the kinetics and extent of virus
Viral Inactivation Issues in Aseptically Processed Parenterals 4011

inactivation. Thus, the process variables evalu- processes should contain other upstream orthogonal
ated during virus clearance studies must be care- steps to assure broad and effective virus reduction.
fully selected and defined since they may alter Regardless of the virus reduction capacity of a manu-
the efficacy of the step for virus reduction. facturing process, however, absence of virus in
2. The results from the albumin studies demon- plasma-derived products can never be guaranteed
strated that virus selection (e.g., HM175/18f because the amount of contaminating virus in source
and HM175/24a) and preparation (e.g., chloro- material is generally not known and the presence
form extracted and untreated virus) were just as and/or appearance of new unknown viruses cannot
important as the process conditions (e.g., protein be excluded.
concentration and temperature) in determining Manufacturing processes have evolved dramati-
the outcome of virus reduction during pasteuri- cally over the last few years. In the late 1980s, 76%
zation. Since laboratory adapted viruses and of hemophiliacs were HCV positive,[40] and between
naturally occurring viruses may differ in their 1979 and 1985, approximately 50% of hemophiliacs
sensitivity to physico-chemical treatments, the had acquired HIV from plasma-derived FVIII.[41]
results observed with model viruses must be care- Since then, however, most U.S.-licensed plasma deriva-
fully interpreted before they can be extrapolated tives have not transmitted HBV, HCV, or HIV as a
to relevant viruses of concern. result of improvements in donor screening and test
3. Certain process steps may be easier to model methods, and the inclusion of effective upstream
than others and duplicating the freeze dry/dry virus-reduction and terminal virus-inactivation steps
heat step at small scale is very difficult. The in manufacturing processes.[18] Residual risks of virus
conclusions drawn from virus clearance studies transmission from plasma-derived products are now
are reliable only when the appropriateness of largely associated with non-enveloped viruses.[42] Thus,
the small-scale models can be demonstrated. the need for additional terminal or upstream virus
During the freeze dry/dry heat step of inactivation/removal steps still exists, but the current
KoateÕ-DVI, virus reduction was dependent on challenge is to develop cost effective methods against
moisture levels so even the formulation of stop- physico-chemically resistant non-enveloped viruses,
pers, which could impact the absorption of such as human parvovirus B19.
water during autoclaving and its release to
freeze dried material, must be considered and
tested.
4. Formulation studies with Protein G suggested ACKNOWLEDGMENTS
that the addition of virus could affect the com-
position and freeze dry/dry heat treatment of The authors thank M. Fournel, J. Wang, and S. F.
spiked product intermediates. As specified in Chao for their comments on the manuscript. They
guidance documents for virus clearance stu- are also grateful to K. Dawson, L. Franks, E. Land-
dies,[1,2] the amount of virus spiked into starting reth, S. Nicosia, and H. Renfrow for expert technical
test materials should be as high as possible so assistance.
that the maximum virus reduction capacity of
a production step can be determined. In many
clearance studies, the volume of the virus added
may be as high as 10%. Since virus reduction REFERENCES
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Virtual Screening
Tı́mea Polgár
Gedeon Richter Ltd., Budapest, Pest, Hungary

György M. Keserü
Computer Assisted Drug Discovery and HTS Unit, Gedeon Richter Ltd.,
Budapest, Pest, Hungary

INTRODUCTION definition of the screening library in a suitable format.

In vitro HTS campaigns are then typically performed in
Identification of viable chemical leads is one of the key a highly automated environment and generates huge
tasks of the early stage of drug discovery. Historically, amount of data that should be stored, treated, and ana-
the lead discovery phase was mainly supported by lyzed. In these contexts, VS techniques can be considered
in vivo experiments that usually resulted in compounds as ‘‘in silico’’ analogues of in vitro HTS technologies
with acceptable efficacy and suitable pharmacokinetic (Fig. 1). After defining the target, VS also requires the
profile at least in animal models. On the other hand, development and optimization of the prediction tech-
however, lead optimization was slow and rarely suc- nique (i.e., the in silico analogue of the assay) applied
cessful that gave significant role to serendipity. Because for affinity predictions. Databases that serve as screening
in vivo based, purely intuitive approaches yielded library should be preprocessed to use in VS. The screening
hardly predictable results; drug discovery faced a para- process itself is performed in the computer memory and
digm shift in the early 1980s. Limitations of animal should, therefore, be fully automated while generating
models and the lack of the molecular mechanism were data similar to HTS datasets both in quantity and quality.
found to be critical factors when analyzing attrition In addition to similarities between the procedures,
rates that initiated research groups to introduce in HTS and VS have a common conceptual framework
vitro tests at the frontline of discovery programs. as well. Both methods have limited accuracy that is
Application of rational approaches was also facilitated compensated by the number of compounds investi-
by in vitro screening supplying reliable datasets for the gated. VS and HTS, are rather classification techniques
first time for computer aided drug discovery (CADD) that separate actives from inactives, than a method of
methods. In vitro assays serve the basis of structure- choice when quantifying biological affinity. These
activity relationship that drives medicinal chemistry dur- methods could identify candidates rather than vali-
ing active-to-hit and hit-to-lead processes. Dramatic dated hits. Some of the promising candidates could
developments in molecular biology, detection methods, never be validated, these are false positives. Both
computer technology, and robotics made high-through- high-throughput technologies surely miss some active
put in vitro screening (HTS) to be a characteristic tool of compounds that are called ‘‘false negatives.’’ Although
lead discovery. In the 1990s, the brut-force HTS was the every reasonable effort should be done to minimize
ultimate technology of lead discovery. Today, however, false positive and negative rates, the common philo-
it became clear that productivity of lead discovery could sophy behind these techniques suggest that if we iden-
not be increased by screening more compounds against tify even a single interesting compound, then false

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more targets even in more and more sophisticated positives and negatives can be tolerated. Finally,

Virtual
assays. In addition to limitations in target validation, candidates identified by HTS or VS should be experi-
assay development, compound and data quality and mentally validated before starting active-to-hit and
despite ultra high throughput, the number of available, hit-to-lead processes. The most characteristic differ-
or ever synthesized compounds is still negligible relative ence between HTS and VS is that the former identifies
to the drug-like chemistry space. Random screening the candidates experimentally, while the latter derives
should, therefore, be replaced by methodologies that them computationally. The computational strategy
combine the capacity of HTS approaches and the applied in VS suggests that: (i) it could be more effec-
rational basis of CADD techniques. Virtual screening tive in time, resource, and cost; (ii) it could explore a
(VS) methods exemplify this idea, as high-throughput significantly larger part of the drug-like chemistry
CADD tools are capable to investigate huge compound space; and (iii) it could yield higher hit rate than ran-
collections in reasonable time and cost. dom screening. Although these potential advantages
Application of the HTS technology requires the selec- suggest VS being more favorable than HTS, compara-
tion of the target, the development of the assay, and the tive studies demonstrated that these approaches are
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-120042127
Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved. 4013
 4014 Virtual Screening

violate more than one rule out of the followings:

molecular weight (MW) < 500, calculated log P
(clogP) < 5, number of hydrogen bond acceptors
(HBA) < 10, and number of hydrogen bond donors
(HBD) < 5. These ranges of properties define a chemis-
try space containing molecules without serious absorp-
tion, distribution, metabolism, and elimination (ADME)
problems that are expected to be orally bioavailable.
This chemistry space is usually referred as drug-like
space and its content is called drug-like molecules.
Consequently, drug-likeness of compound libraries
(both virtual and physical) can be easily investigated
by assessing their members by ROF. In addition to
properties included to ROF, Oprea proposed further
limitations in features when searching for drug-like
Fig. 1 Basic steps in virtual screening and HTS. molecules.[2] His analysis suggested that the number of
rotatable bonds (RB) should be lower than eight while
the number of rings ideally smaller than four. Kelder
rather complementary than competitive. In one hand, et al.[3] introduced polar surface area (PSA) as a relevant
VS techniques could be useful when designing target descriptor for predicting oral bioavailability. PSA
specific screening libraries. On the other hand, distinct was defined as the sum of the van der Waals surface
structural classes identified by VS and HTS are clearly of polar atoms including oxygen, nitrogen, and sulfur
the observations that most straightforwardly support with attached hydrogens. PSA showed significant
complementarities between VS and HTS. While the impact on membrane penetration including intestinal
underlying disciplines of HTS are bioinformatics (PSA < 140 A2)[4] and blood–brain barrier permeabil-
and biochemistry, VS is based on cheminformatics ities (PSA < 80 A2).[5,6] As PSA can be approximated
and computational chemistry. In fact, there are from the 2-D structure, these approaches are now
computational approaches–most of them are CADD considered as filtering methods for VS of large com-
methods–already applied in drug discovery that were pound libraries. Although purely statistical approaches
transformed to VS tools. Similar to CADD techniques are still dominating on this field, Gillet, Willett, and
VS methods can be divided into ligand-based and Bradshaw[7] used a genetic algorithm (GA)-based
structure-based approaches. Ligand-based techniques weighting scheme for property and shape descriptors
operate exclusively in the small molecule space using when discriminating between drugs and non-drugs.
chemical and biological information encoded in known The concept of lead-likeness has been established by
active compounds to identify new candidates with Hann[8] and Oprea,[9] analyzing lead molecules and
similar properties. Structure-based methods require corresponding drugs. Comparative studies on leads
the three-dimensional structure (3-D) of the target and drugs revealed that leads are typically less complex
protein that is used for docking small molecules into molecules than drugs. Consequently, their physico-
the binding site. Evaluation of binding modes and chemical properties should differ significantly from
protein-ligand interactions finally allows ranking of that of the drugs. In fact, effective leads have lower MW
docked compounds by their binding affinity. This entry and clogP, have a smaller number of rings (RNG), and
summarizes the background of both approaches, gives RB. The original ROF was therefore modified to rule-
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case studies for supporting practical applications, and of-three (ROT: MW < 300, clog P < 3, H-bond donors
Virtual

collects success stories to demonstrate capabilities. (HDO) < 3, rotatable bond (RTB) < 3) by Astex
reflecting to the reduced complexity of leads.[10] Based
on these findings, Leach et al. established a filtering
scheme of MW < 350, RTB < 6, number of heavy
DATABASE FILTERING atoms < 22, HDO < 3, high-activity moleucle (HAC)
< 8, clogP < 2.2 when selecting compounds for reduced
Physicochemical Filters complexity screening.[8] Surveying medicinal chemistry
literature Oprea and coworkers[11] have drawn less
One of the early physicochemical filters is the famous strict conclusion, when formulating the criteria of
rule-of-five (ROF) introduced by Lipinski et al.[1] lead-likeness MW < 460,
4 < Log P < 4.2, RTB
ROF is based on the distributions of easily accessible < 10, RNG < 4, HDO < 5, HAC < 9.
and interpretable physicochemical properties obtained As similar to HTS, the basic objective of VS is
from known drugs. Compounds fulfilling ROF do not to identify interesting starting points for medicinal
Virtual Screening 4015

chemistry programs. We believe that lead-likeness comprising filters for reactive functional groups,
would be a useful concept for filtering compound unsuitable leads (i.e., compounds which would not be
libraries before more sophisticated approaches. Never- initially followed up), and unsuitable natural products
theless, it should be emphasized that limits applied for (i.e., derivatives of natural product compounds known
both drug-like and lead-like compounds are based on to interfere with common assay procedures). Some of
statistical analyses of known examples. Although these these filters are depicted in Table 1.
simple filters demonstrated significant classification Finally, soft filters represented by a GA trained to
accuracy when discriminating drugs and non-drugs, identify unsuitable compounds that the other filters
leads and drugs–their fully empirical nature warns would fail to find. This algorithm scores compounds
against unconditional applications. for drug-likeness, relative to a training set classified
by medicinal chemists.
Functional Group Filters Andrews and coworkers[14] used a set of 200 drug
molecules to derive a set of ‘‘intrinsic binding ener-
Functional group filters are mainly utilized to remove gies’’ for the 10 functional groups shown in Table 2.
unstable, reactive, toxic, or otherwise unsuitable com- The inherent binding affinity of compounds was then
pounds from compound libraries. The rapid elimina- estimated by summing the intrinsic binding energies
tion of swill (REOS) method introduced by Vertex and subtracting an entropic factor.
was the first realization of this concept.[12] REOS effec- Muegge, Heald, and Brittelli described a functional
tively combines physicochemical filters with a set of group filter to discriminate between drugs and non-
functional group filters. Databases are first subjected drugs.[15] The authors assigned a score to each molecule
to property filtering similar to ROF that is followed that is based on the presence of fragments typically
by checking a set of rules based on the presence of found in drugs (Table 2). Each non-overlapping
functional groups expected to be problematic. Some drug-like fragment counts one point in this scheme.
examples of these rule-based functional group filters Molecules with a score between two and seven are
are illustrated in Fig. 2. It is important to note that classified as drugs, otherwise they are classified as
REOS allows the user to customize each functional non-drugs. As it was noted that CNS active com-
group filter as well as the set of rules applied. pounds are relatively small and typically contain only
Hann et al.[13] used a similar, multilevel approach a single pharmacophoric group, they defined a second
when pooling compounds for HTS. These authors filter. This filter ensured that compounds containing a
applied three types of filters. Basic filters were used single drug-like functional group could only be classi-
to remove molecules with non-drug-like features. Next fied as drugs if they contain one of the distinguished
functional group-based hard filters were utilized groups indicated with an asterisk in Table 2.

Topological Filters

It is generally accepted that structural similarity to

known drugs significantly increases the drug-like
character of compounds. Therefore, molecular topology
of known drugs in comparison to non-drugs served as
suitable starting point for a number of approaches.
One group of methods utilized neural networks for dis-
criminating between drugs and non-drugs. Compounds

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in drug databases such as World Drug Index (WDI),

Virtual
Comprehensive Medicinal Chemistry (CMC), and
MDL Drug Data Report (MDDR) and databases of
chemical suppliers (typically Available Chemical Direc-
tory; ACD) were represented by structural descriptors.
Sadowski and Kubinyi[16] reported the application of
Ghoose–Crippen atom types as input neurons, while
the output neuron was set to one for drugs and
zero for non-drugs when training the net by 5000
WDI and 5000 ACD compounds. The trained net was
able to classify about 80% of test compounds.
Murcko and coworkers reported a similar approach
based on drugs collected from CMC and non-drugs
Fig. 2 Examples of functionalities filtered out by REOS. from ACD.[17] Molecules were represented by 166 binary
 4016 Virtual Screening

Table 1 Functional group-based filters suggested by Hann et al.[13]

Reactive groups Unsuitable leads Unsuitable natural products
Reactive alkyl halides Disulfides Quinones
Carbazides Aziridines Saponins
Sulphate esters Thiols Polyenes
Sulphonates Cyanamides Squelastatin derivatives
Acid anhydrides Thiocyanates Cytochalasin derivatives
Isonitriles Four-membered lactones Monensin derivatives

ISIS keys and the authors added further physico- number of heavy atoms, and total charge. Predictive
chemical descriptors, including MW, number of HBD power of the trained neural network was first assessed
and acceptors, number of RBs, aromatic density, in the test set. Using a threshold of 0.15 as a criterion
clog P, and one additional descriptor reflecting the to discriminate between drugs and non-drugs, the
degree of branching. A Bayesian neural network neural network was able to classify 88% of the MDDR
(BNN) was trained using 3500 molecules from both drug-like set and ACD databases correctly. External
the CMC and the ACD databases. The trained net- validation on lead-like compounds predicted 75% of
work was then tested on CMC and ACD compounds these molecules as drug-like. Although neural network
that were not included in the training set. A subset of based methods are extremely fast and successfully
the MDDR library was used for external validation. identified the majority of known drugs, these models
The classification accuracy of both CMC and ACD are hardly interpretable for medicinal chemists and,
approached 90%, while external validation revealed therefore could rather be used as high-throughput
78% of the MDDR compounds being drug-like. filters than driving chemistry to the drug-like space.
Frimurer et al.[18] also reported a neural network Wagener and Geerestein[19] applied recursive
based approach trained on a larger dataset. These partitioning to distinguish of drugs and non-drugs.
authors partitioned the MDDR database into drug- Analyzing tolerated and non-tolerated functional
like (compounds that have progressed to at least groups in both WDI and ACD databases the authors
Phase I of clinical trials) and lead-like (compounds were able to recognize almost 75% of drug-like mole-
labeled as ‘‘Biological Testing’’) sets. Diversity selec- cules in MDDR and CMC databases.
tion from these sets resulted in 4400 MDDR drug-like Limited number of structural frameworks represented
compounds (3000 training and 1400 test compounds), in known drugs can also be used for the identification of
and 90,000 ACD compounds (60,000 training and drug-like compounds. In their pioneering work, Bemis
30,000 test molecules) dissimilar to the drug-like set. and Murcko analyzed shapes of existing drugs and
The 60,000 lead-like MDDR compounds were used identified drug-like molecular frameworks.[20] A graph-
exclusively for external validation. All compounds theoretical approach was used to decompose molecules
were represented by 77 CONCORD atom types and into fragments (Fig. 3). Rings and linkers together form
three further descriptors including the number of atoms, the framework, while acyclic side chains were removed.

Table 2 Selected functional groups used in scoring

schemes developed by Andrews[14] and Muegge[15]
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Virtual

Andrews scheme Andrews score Muegge scheme

Carboxylate 8.2 Amine
Phosphate 10 Amide
N+ 11.5 Alcohol
N 1.2 Ketone
OH 2.5 Sulfone
CO 3.4 Sulfonamide
O or S 1.1 Carboxylic acid
Halogens 1.3 Carbamate
C (sp2) 0.7 Guanidine
Fig. 3 Decomposition scheme for the identification of drug-
C (sp3) 0.8 Amidine
like frameworks. (From Ref.[20].)
Virtual Screening 4017

query using chemical similarity principles. During a simi-

larity search, the query molecule is compared to members
of the database and a similarity measure is calculated
quantifying the similarity between the query and each
molecule in the screened database. For 2-D similarity
searches, the query structure and members of the screened
database are typically represented by molecular finger-
prints that encode molecular structure and properties in
binary format (Fig. 5). Each bit of the fingerprint detects
the presence or absence of molecular fragments or
connectivity pattern, can encode descriptor value ranges
or binary transformed descriptors. Fingerprints can be
divided into two major groups: structural keys and
hashed fingerprints. Structural keys associate a specific
descriptor (structural fragment or property) with each
Fig. 4 Retrosynthetic reactions defined in RECAP.
bit, while hashed fingerprints assign overlapping bit
segments to descriptors. Generation of structural keys
requires a predefined fragment library that collects the
Based on this analysis, the authors concluded that fragments searched in each molecule of the database.
almost the 50% of known drugs in the CMC could be MACCS key (MDL Information Systems) is a typical
described by only the most frequent 32 frameworks. example for a structural key. In contrast, there is no
The diversity of drug-like side chains was found to be fragment library needed for hashed fingerprints. These
similarly low, only 20 different side chains account for fingerprints are derived algorithmically from all possible
over 70% of all the side chains appeared in CMC.[21] linear paths of the limited number of connected atoms.
It is interesting to note that this approach is also Paths define a pattern of atoms and bonds that are used
useful for designing target-based libraries when the to generate a set of bits during the hashing procedure that
topological framework analysis is applied to active typically produces 4–5 bits per pattern. Although hashed
compounds within a target family. One realization of fingerprints are not physically interpretable, they show
this concept is the retrosynthetic combinatorial analy- highly characteristic bit patterns for molecules. Bit strings
sis procedure (RECAP) algorithm that was also vary in length and complexity but Daylight (Daylight
utilized for the analysis of drug-like fragments.[22] Inc.) and UNITY (Tripos Inc.) are the most popular
RECAP first identifies active or drug-like fragments hashed fingerprints used in similarity searches.
using retrosynthetic analysis applying any of the Analysis of molecular similarity is based on the
11-retrosynthetic reactions defined (Fig. 4). In the next quantitative determination of the overlap between
step, the resulting fragments are usually clustered and fingerprints of the query structure and all database
transformed into monomers that can be used for members. As descriptors of a given molecule can be
designing targeted libraries. As fragments were considered as a vector of real or binary attributes, most
extracted from known actives, it is expected that com- of the similarity measures are derived as vectorial
pounds enumerated from the derived monomers will distances. Tanimoto and Cosine coefficients are the
be also active and are synthetically accessible because most popular measures of similarity.[24] Definitions of
of the retrosynthetic approach applied. similarity metrics are collected in Table 3.
A similar method, TOPology-Assigning System In addition to binary descriptors, molecular holograms
(TOPAS) has been reported by Schneider et al.[23] are also useful for similarity searches.[25] Similar to
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Virtual
These authors developed an evolutionary algorithm fingerprint, hologram is a vector that contains numerical
for fragment-based de novo design. This stochastic
method utilizes a set of 25,000 fragments that serves
as building blocks obtained by a rule-based fragmen-
tation procedure applied to 36,000 known drugs.

LIGAND-BASED SCREENING

Similarity Searches

Similarity searching is one of the simplest methodologies Fig. 5 Structural representation by molecular fingerprints and
of ligand-based VS, when screening databases against a holograms.
 4018 Virtual Screening

Table 3 Most popular similarity metrics

Metrics Definition
qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
Pi¼N 2
Euclidean i¼1 ðxiA
xiB Þ

Pi¼N
i¼1 xiA xiB
Cosine qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
Pi¼N 2 Pi¼N 2
i¼1 ðxiA Þ i¼1 ðxiB Þ

Pi¼N
xiA xiB
Tanimoto Pi¼N 2 Pi¼1 Pi¼N
i¼1 ðxiA Þ þ i¼N
i¼1 ðxiB Þ2
i¼1 xiA xiB

xiA: is the ith bit in object A; xiB: is the ith bit in object B.

Fig. 6 Schematic representation of pharmacophore features.

properties, e.g., number of occurrences for atoms or

fragments.
3-D similarity searches utilize 3-D information includ- constraints. Pharmacophore features can be derived
ing shape, steric, and electrostatic properties obtained for from both the structure of the target protein or from
the query molecule and the database screened. High- the collection of known ligands (Fig. 7).
throughput molecular alignment techniques of this kind Structure-based pharmacophores are typically
superpose all database entries onto the query molecule. explored by analyzing binding site interactions formed
FlexS, one of the most popular approaches, keeps the between ligands and protein atoms within the active
query rigid and considers test molecules as flexible when site of the target. In addition to direct structural infor-
offering several alternative superpositions–each scored mation, site directed mutagenesis data are also useful
and ranked by similarity.[26] Genetic algorithm similarity when identifying most important residues responsible
program (GASP) is a superposition tool using GA that for ligand binding. LigandScout uses ligand-bound tar-
handles both the query and the test molecules as flex- get structures to extract structure-based pharma-
ible.[27] Wild and coworkers[28] described an alignment cophore models.[31] Hydrogen bonding features, as
tool utilizing GA-based fitting of molecular electrostatic well as electrostatic and hydrophobic interactions, are
potential fields. In MIMIC, Mestres, Rohrer, and Mag- utilized to derive the most important interactions
giora[29] represented molecules as sets of Gaussian func- within the active site. Starting from the protein
tions, modeling property fields. Alignment is optimized structure, LUDI generates interaction sites that are
and scored assessing the overlap between corresponding preferable to occupy for ligand atoms.[32] This method-
fields. Rapid overlay of chemical structures (ROCS) ology is the basis of the structure-based focusing (SBF)
performs a shape-based superposition using a Gaussian technique available in Cerius2. The identification of
representation of the molecular volume and allows the interaction sites by LUDI is followed by the clus-
to combine 2-D and 3-D similarity.[30] tering of interaction vectors. The obtained clusters
are the starting point for generating the pharma-
cophore hypothesis. POCKET realizes a similar
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Pharmacophore Searches approach to LUDI when identifying grids within the

Virtual

active site to characterize favorable ligand inter-

Pharmacophore mapping actions.[33] Considering the flexibility of the active
site, Carlson et al. suggested a molecular dynamics
Pharmacophore is an arrangement of steric and elec- approach when generating a diverse set of protein con-
trostatic features in the 3-D space that are crucial for formations to develop structure-based pharmacophore
the biological action (Fig. 6). These kinds of feature- models.[34] Reflecting to multiple potential binding
based pharmacophore models were found to be useful modes, geometric constraints between pharmacophoric
for ligand-based VS because they can effectively fish points are here defined as ranges rather than values.
very different chemotypes from virtual libraries. Phar- Pharmacophore constraints can include not only dis-
macophore models usually consist of a number of tance, angles, and dihedrals, but also excluded
pharmacophore points including a group of atoms, volumes; surface volumes and spatial restraints might
or features such as HBDs and HBAs, charged groups, also be used in the UNITY program.[35] Molecular
hydrophobic centers, and corresponding geometric Operating Environment (MOE) also allows queries
Virtual Screening 4019

Fig. 7 Approaches used for pharmacophore mapping.

containing locations of features, chemical groups, or In Catalyst, the random search algorithm used for
shape constraints.[36] the identification of low energy conformers is conduc-
In the absence of structural information on the target, ted with a pooling function that allows the in-depth
pharmacophore could be developed using a set of coverage of the conformational space. Features are
actives that are expected to realize similar binding then identified by analyzing the surface accessibility
modes when interacting with the target. Ligand-based of the actives followed by the definition of the pharma-
pharmacophore generation that is also called as cophore using the absolute coordinates of all confor-
pharmacophore mapping usually consists of three ele- mations. HipHop is useful to generate qualitative
mentary steps. In the first step, the pharmacophore hypotheses, while HypoGen is the method of choice
features, common in all of the actives, are identified. for more predictive quantitative models. The initial
The second step involves the generation of putative model is based on the features of the two most active
bioactive conformations that might be explored by molecules that serve as a starting point to generate all
the actives. Finally, alignments of these conformations the possible pharmacophores. Models are then evalu-
are prepared that ensure matches of pharmacophore ated by analyzing the overlay between the pharmaco-
features. Identification of possible bioactive confor- phore hypotheses and the geometric arrangement of

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mations is clearly the greatest challenge in pharma- features in all conformations of the actives. For quan-

Virtual
cophore mapping. There are two basic strategies to titative models, Catalyst assigns weight descriptors to
achieve this goal. The first one generates only low each feature that correspond to its importance in
energy conformations for all of the actives and then explaining the biological activity. Disco[39] and Disco-
searches for common features. The other one, however, Tech,[35] both characterize actives by ligand and site
enumerates all possible conformations of each ligand points. Ligand points include HBDs and HBAs, and
and evaluates the orientation of pharmacophore hydrophobic and charged features. Site points are
features previously identified. generated on the basis of complementarity principles
Although pharmacophore mapping can be com- using the atomic coordinates of the corresponding
pleted manually in simple and straightforward cases, ligand atom. Similarly to the Catalyst approach, a set
most of the applications require the automated gener- of low energy conformers is first generated that are
ation of pharmacophore. There are several software aligned as rigid objects. Alignment of the conformers
suites available for this purpose. HipHop[37] and Hypo- represented by interfeature distances to a reference
Gen are both featured by the Catalyst package.[38] molecule having the fewest conformation is carried
 4020 Virtual Screening

out by a click detection algorithm. Disco produces a scaffold hopping or lead hopping that is not limited
ranked list of hypotheses that contains all of the to lead discovery programs but might contribute to
common features identified in actives. Alignment param- back-up/follow-up strategies.
eters calculated by Disco drive the users to select Pharmacophore searches are usually realized in two
good quality pharmacophore maps. In GASP,[27] the steps. Search algorithms first check the availability of
conformational analysis of actives takes place on the pharmacophore features encoded in the query and next
fly that is followed by the assignment of pharmaco- they evaluate the overlay of the spatial arrangement of
phore features including both the ligand points and site features and the query. The fastest way of the second
points. The molecule with the least number of features step is matching database entries as rigid objects. As
serves as reference. The alignment procedure is carried flexible molecules can adopt a number of different
out by a GA utilizing chromosomes that encode low energy conformations, this set of conformers
dihedrals of RBs in all actives and mapping of the should be precomputed before screening. Another
reference’s pharmacophore features to other actives. option for rigid searches is the on the fly generation
The fitness function generates conformations for each of conformers that is followed by the subsequent struc-
active that are then fitted to the reference using the tural alignment. Conformational flexibility is more
mapping information. The best possible overlay is explicitly considered in flexible 3-D searches that, how-
achieved by calculating the internal van der Waals ever, require significantly higher resources. Pharmaco-
energy of each molecule, the number, and similarity phore queries that include constraint ranges could
of the overlaid features in combination of the volume partially compensate conformational changes occur-
integral of the overlay. ring upon ligand binding. There are a number of codes
available for database searching. 3DSEARCH[40]
Pharmacophore search divides the searches into two parts, a fast prescreen
using an inverted key system and a slower atom-
The pharmacophore model obtained by the mapping by-atom geometric search using the Ullman algorithm.
procedure allows 3-D database searching (Fig. 8). Features to handle angle/dihedral constraints and to
Database searching can identify actives in different take into account ‘‘excluded volume’’ are implemented
chemotypes relative to those utilized for pharmaco- as part of the geometric search. Catalyst utilizes both
phore generation. This procedure is often termed as multiconformer databases and the on the fly conformer
generation. ChemDBS-3D generates low energy con-
formers that are filtered by conformational rules.[41]
Root mean square (RMS) deviation from the pharma-
cophore constraint can be minimized by torsional
minimization. The flexible 3-D search available in
UNITY realizes this concept based on the Directed
Tweak algorithm[42] that could match the constraints
of the query. A recent review by Langer and Wolber[43]
compares these technologies when used for pharmaco-
phore searches.

Case Study: Ligand-Based VS against

Kv1.5 Potassium Channel
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Peukert et al. reported VS experiments using different

ligand-based techniques against the voltage-dependent
potassium channel Kv1.5.[44–46] In their first attempt,
the authors utilized similarity searching to identify
new actives.[44] A potent indane derivative first pub-
lished by Eli Lilly was used as a query in a 2-D simi-
larity search performed in UNITY. Compounds in
the corporate collection were represented by 2-D
UNITY fingerprint and the fingerprint of the query
was compared to that of the library members. Com-
pounds with Tanimoto coefficient larger than 0.8 were
considered to be similar and was subjected to biologi-
Fig. 8 Strategies used in pharmacophore searches. cal testing. This protocol led to the identification of a
Virtual Screening 4021

CI
O
O N CONH2
S O N
N F 3C S
O N O
O

Initial hit Lead

Fig. 9 The initial hit from similarity searching was converted a viable lead.

structurally novel hit with moderate potency (9.5 mM) subsequent UNITY 3-D flexible search. The perform-
and limited chemical stability (Fig. 9). Suboptimal ance of this ligand-based VS tool was first evaluated
physicochemical properties and stability prompted against an in-house set of known Kv1.5 potassium
the authors to replace the central naphthalene moiety channel blockers. This test revealed that the method-
that resulted in the discovery of a good quality lead ology was able to retrieve 58% of known actives. VS
with improved potency (4.8 mM). Optimization of this was performed on the Aventis compound collection
lead was supported by solid phase parallel synthesis and identified 4234 virtual hits that were first subjected
that enabled the authors to identify a set of highly to filtering. Compounds with reactive or non-tolerated
potent and selective Kv1.5 potassium channel blockers. features regarding the Kv1.5 potassium channel inhib-
Structure-activity information gained for this com- itory activity, with non-drug-like character, as well as
pound series and also that obtained in a parallel lead known blockers were removed during this process.
optimization program allowed the development of a The final set of 1975 compounds was submitted to hier-
pharmacophore model for Kv1.5 potassium channel archical clustering that resulted in 27 clusters in total.
blockers.[45] Seven potent molecules from each series were Representatives of the available 18 clusters were tested
used to derive the model that consists of three hydro- against Kv1.5 potassium channel that identified one
phobic centers in a triangular arrangement (Fig. 10). structurally novel compound with an IC50 of 5.6 mM
The seven compounds were first subjected to an (Fig. 11). This hit was considered as a starting point
exhaustive conformational search utilizing the Monte for optimization that led to the identification of a series
Carlo (MC) Multiple Minimum algorithm as available of compounds with remarkable activity (best IC50 was
in MacroModel. The resulting conformers were then found to be 0.5 mM) and acceptable pharmacokinetics.
clustered using distances between the potential phar- In the final round, the authors reported the develop-
macophore features. A minimum energy conformation ment of a structure-based pharmacophore that was
in each of the clusters was used as input to DISCO also used for VS.[46] To achieve this goal, a homology
algorithm. DISCO models were visually inspected model of the Kv1.5 potassium channel has been
and one of them was used as a 3-D query in the developed. The pore domain of the target protein

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Virtual

Fig. 10 Definition of the pharmacophore

query for the Kv1.5 channel. Representatives
of the two compound series (A and B)
werealigned(b)toderivethepharmacophore(a).
(Copyright is permitted by Elsevier).
 4022 Virtual Screening

Fig. 12 Homology model of the Kv1.5 pore. Amino acids at

the extracellular site is colored yellow, while those at the
internal site is in magenta (A); Amino acids forming the
internal site. Amino acids found to be essential for Kv1.5
blocking activity indicated gray, while others are green.
GRID isocontours are blue, red, and orange for the H-bond
donor, acceptor and hydrophobic probes, respectively (B);
Structure-based pharmacophore derived from the GRID
analysis of the internal site colored according to correspond-
Fig. 11 The structurally novel lead identified by the phar- ing iso-contours. The excluded volume is colored yellow (C).
macophore search. Lead optimization was focused to regions (Copyright is permitted by the Copyright Office of the
indicated. American Chemical Society).

was built using the crystal structure of the bacterial maps in addition to the exclusion volume defined by
KcsA channel. A single subunit of the Kv1.5 channel protein atoms were used to define the pharmacophore
was generated by the COMPOSER program as imple- query (Fig. 12). This structure-based pharmacophore
mented to SYBYL software that was used to create the was utilized for a UNITY Flexsearch-based VS that
tetrameric structure arrangement of the target protein. resulted in 3102 hits in total. Drug-like filters, as well
The resulting model was then subjected to structural as partial match constraints (at least 3 out of the 12
refinement using a two-step minimization protocol features), were applied. Compounds with reactive or
and it was validated by a number of protein analysis non-tolerated features regarding the Kv1.5 potassium
tools including PROCHECK, WHATCHECK, and channel inhibitory activity were removed. After visual
MATCHMAKER (Fig. 12). The binding site was inspection of the hits by medicinal chemists, the
identified by applying geometrical criteria on the authors identified 244 compounds that were subjected
validated model using the putative active site for to in vitro screening. Biological evaluation of the hits
spheres (PASS) algorithm. Visual inspection of PASS revealed 19 actives in total including 5 compounds with
results led to the selection of a binding site that has an inhibition concentration (IC50)[47] under 10 mM,
been further characterized by the GRID force field. while the best compound having an IC50[47] of 9 mM.
Pharmacophore features were derived by the calcu- As all of the ligand-based efforts described in these
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Virtual

lation of molecular interaction fields using the hydro- papers were carried out on the same screening library
phobic, the amide nitrogen, and the carbonyl oxygen (i.e. the Aventis compound repository), hit rates of
probes. Local minima of the fields were visually different approaches could directly be compared.
inspected and the selected minima of GRID energy (Table 4).

Table 4 Success rates of ligand-based virtual screening for Kv1.5 potassium channel
Method Ncomp Nact Hit rate(%) Best IC50 (lM) Nclass
2-D similarity search 75 2 2.7 7.7 2
Ligand-based pharmacophore 18 1 5.5 5.6 1
Structure-based pharmacophore 244 19 7.8 0.9 5
Ncomp, number of compounds selected for in vitro screening; Nact, number of active compounds; Best IC50, IC50 values of the most active
compound; Nclass, number of chemical classes identified.
Virtual Screening 4023

Based on this comparison, one can conclude that lead molecules. The investments in structural biology
pharmacophore search–both with ligand-and structure- prompted the improvement of structure-based VS.
based pharmacophores–outperformed 2-D similarity Structure-based VS involves explicit molecular docking
search. Although the two latter techniques gave similarly of each ligand into the active site of the target protein.
good result in hit rate, the authors found no overlap Hereby, a predicted binding mode is produced, and
between their hit lists. Comparing the number of chemi- then the quality of the fit of the molecule in the
cal series identified it is clear that screening by the struc- active site is scored. This scoring information is used
ture-based pharmacophore hypothesis outperformed the to rank the compounds and only the top solutions
ligand-based approaches. It should be noted, however, can be investigated further. A pictorial overview of
that the effectiveness of different VS protocols obviously the docking workflow is shown in Fig. 13.
depends on the target and the quantity and quality Without the need of completeness, we discuss the
of structural and structure-activity relationship (SAR) most state-of-the art structure-based VS that are gener-
information actually available. The multiple chemotypes ally used at pharmaceutical industries during the drug
identified by different strategies suggest the usefulness of discovery process. Our attempt was also to give a pictur-
parallel VS programs against the same target but with esque overview on the structure-based VS process and
different approaches. show its effectiveness or its failures through a case study
and some success stories during the CADD processes.

Success Stories
Preparation of Protein Structures
Success stories that demonstrated the capabilities of
both approaches in the CADD technique have been The first step in the structure-based VS process is to
described in Table 5. obtain the coordinates of a protein. Generally, struc-
tures solved by X-ray crystallography or nuclear
magnetic resonance (NMR) are used, but protein
STRUCTURE-BASED VS structures, which have difficulties in the crystallization
process e.g., membrane proteins such as G-protein
The functions of drug molecules and protein targets coupled receptors (GPCR) can be modeled based on
are regulated by the principles of molecular recog- homology.[63] Currently, 3-D structure information
nition. The rational drug development requires the can be generated for up to 56 % of all the known
understanding of molecular recognition in terms of proteins. However, there is considerable controversy
structure and energetics.[59] Structure-based VS tools concerning the real value of homology models for
study the binding between the ligand and protein struc- structure-based VS. Recently published results on
tures with respect to structural and energetic consid- ligand-supported homology modeling might be able
erations. In the earlier era of the structure-based to provide reasonable quality homology models that
drug discovery process, the low resolution of protein are suitable for structure-based VS.[64,65]
structures and poor computational power hindered The definition of the active site is needed for docking
the rapid improvement of the method. Today, these into a target protein because scanning the entire surface
techniques have experienced a comeback because dras- of the protein would hardly be feasible with most of the
tic changes occurred. currently used docking algorithms. On the other hand,
Genomics have resulted in a huge number of poten- ignoring biochemical information or structural data on
tial therapeutic targets that are available for investi- the active site is unreasonable, although, in some cases

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gation. However, learning the sequence of a genome information about binding sites is not available. These

Virtual
is a long way from understanding its biological func- cases-binding sites can be identified by algorithms detect-
tion. Predictions of protein function can be attempted ing geometric cavities,[66,67] or algorithms based on
from the knowledge of the structure alone or other physicochemical and geometrical characterization.[68]
additional information can be used to gain information The preparation of the active site depends on the
about functional predictions. In many respects, it is docking tools being used. This step involves the
still early days for seeing the fruits of this process in addition of hydrogen atoms avoiding atomic clashes;
the products offered on the market by pharmaceutical assignment of appropriate protonation states of titrat-
companies. Therefore, there are still large investments able residues and correct tautomers of histidine resi-
in functional genomics,[60–62] HTS methodologies, dues, and involvement of structural water molecules
combinatorial chemistry, predictive ADME methods, in the binding cavity.
and structural biology. The conformational flexibility in the active site has
VS, and structure-based VS, has emerged as an inex- to be evaluated. Commonly used docking methods,
pensive and straightforward method for identifying however, are able to consider the flexibility to a limited
 Virtual
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4024

Table 5 Summary of recently published ligand-based virtual screening studies indicating hit rates (%) and best IC50 values
Target Screening database Program Method Selected Hit rate Hits (best IC50) References
Kv1.5 potassium channel Aventis in-house (N.R.) UNITY Similarity search 75 2.7 2 (7.7 mM) [44]
Kv1.5 potassium channel Aventis in-house (N.R.) DISCO/UNITY Ligand-based pharmacophore 18 5.5 1 (5.6 mM) [45]
Kv1.5 potassium channel Aventis in-house (N.R.) COMPOSER/UNITY Structure-based pharmacophore 244 7.8 19 (0.9 mM) [46]
Rhinovirus 3C protease WDI (47,660) HipHop/Catalyst Ligand-based pharmacophore 2467 0.1 3 (N.R.) [48]
Dopamine transporter NCI 3D (206,876) Chem-X Ligand-based pharmacophore 70 62.8 44 (0.5 mM) [47]
Sigma1 WDI (48,405) HypoGen/Catalyst Ligand-based pharmacophore 389 6.4 25 (1.3 nM) [49]
Muscarinic M3 AstraZeneca in-house DISCO/UNITY Ligand-based pharmacophore 172 1.7 3 (213 nM) [50]
(N.R.)
HIV-1 integrase NCI 3D (206,876) Chem-X Structure-based pharmacophore 340 2.9 10 (1.5 mM) [51]
HIV-1 integrase ChemBridge (150,000) HipHop/Catalyst þ Ligand-based pharmacophore þ 110 18.0 20 (11 mM) [52]
GOLD docking
Urotensin II Aventis in-house (N.R.) HypoGen/Catalyst Ligand-based pharmacophore 918 1.2 11 (400 nM) [53]
MCH-1 Commercially available FlexS 3-D similarity search 526 2.1 11 (55 nM) [54]
(615,000)
VLA-4 ACD (8,624 reagent) HypoGen/Catalyst Ligand-based pharmacophore 12 25.0 3 (1 nM) [55]
CDK2 Pharmacopeia in-house HypoGen/Catalyst Ligand-based pharmacophore 4478 0.8 39 (N.R.) [56]
(117,180)
Farnesyl transferase Schering-Plough in-house HypoGen/Catalyst Ligand-based pharmacophore 330 1.5 5 (0.7 mM) [57]
(N.R.)
mGluR5 Asinex Gold (20,000) CATS3D/MOE 3-D similarity search 27 33.0 9 (12 mM) [58]
The table is provided to allow comparison of searching schemes with respect to the program, method, and screening database. N.R.: not reported.
Virtual Screening
Virtual Screening 4025

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Fig. 13 The overall picture of the structure-based docking workflow.

extent. Therefore, the selection of the target structure is procedures, the representation of atomic coordinates
a crucial point in the docking procedure, so target- is not a practical choice. Thus geometric space descrip-
binding sites should be as representative as possible, tors[69,70] alone or combined with physicochemical
mimicking a real and general binding conformation. descriptors, sphere images[71] and interaction points/
surfaces[72] are commonly used instead.
Otherwise, a complete representation of ligands in
Docking Algorithms atomic coordinates is definitely feasible. The central
problem in ligand handling is the flexibility. Two gen-
In spite of the fact that only a specified part of the erally used strategies can be applied: whole molecule
target protein is considered during the docking and fragment-based methods. Earlier rigid docking,
 4026 Virtual Screening

when the whole molecules are docked, was commonly Systematic methods (incremental ligand
used. An extension of rigid body docking is the incor- building and conformational search)
poration of conformational flexibility. Historically, the
DOCK algorithm addressed rigid body docking using These kinds of algorithms make an attempt to explore
a geometric matching algorithm to superimpose the all the degrees of freedom but face the problem of com-
ligand onto a negative image of the binding pocket. binatorial explosion:
Important features that improved the algorithm’s abil-
ity to find the lowest energy-binding mode, including Y Y 360
N nincrement
force field-based scoring, on the fly optimization, an Nconformations ¼
yi;j
improved matching algorithm for rigid body docking, i¼1 j¼1

and an algorithm for flexible ligand docking, have been

added over the years. This multiconformer docking yi;j
the size of incremental rotational angle j
can be an effective tool, however, for flexible molecules
for bond i
that have significantly higher number of conformers
it may not be straightforward. Fragment-based meth- N
the number of rotatable bonds
ods provide an alternative solution for the docking
problem; molecules are dissected into fragments that Thus ligands are often incrementally built within the
can be docked individually, either separately or active site. The principle of the FlexX algorithm is that,
incrementally. physicochemical properties provide the most useful
Now there is a driving force to treat protein flexible information for ligand placement. Once a set of favor-
because ligands requiring larger conformational able placement of the base fragment (the core part of
changes in the active site upon binding cannot be the ligand from where the incremental construction
placed correctly by these methods. The available starts) has been computed, the ligand building can be
approaches have recently been reviewed.[73,74] The started. The incremental construction is formulated
degree of the applied flexibility varies in a wide range. as a tree search (Fig. 14) problem.
Small adjustments of rigid structures, explicit side As the search tree grows exponentially, a complete
chain flexibility, or the use of protein ensembles are search is definitely not feasible to calculate. Instead
very popular methods, however, the full consideration the binding energy values, approximated by the scores,
of flexibility still remains as the domain of the mole- which are represented by the interior nodes, can be
cular dynamical simulations. used to guide the search. Conversely, DOCK 4.0 algor-
This subsection discusses the search algorithms to ithm divides ligands into rigid (core fragment) and
treat ligand flexibility and to some extent protein flexi- flexible parts (side chains); the core parts are docked
bility. A representative set of the currently reviewed prior to the flexible parts. The steric complementarity
docking[75–78] programs is summarized in Table 6. is the guideline during the ligand construction.

Table 6 A representative summary of commonly used docking programs

Program Method References
DOCK Geometric matching, incremental ligand building [71]
FlexX Geometric matching, incremental ligand building [79,80]
SLIDE Geometric matching, conformational ligand ensembles [81]
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AutoDock Energy-driven/stochastic MC simulated annealing [82]

Virtual

ICM MC minimization, Pseudo-Brownian sampling and [83]

local minimization
QXP MC minimization [84]
MDD Molecular dynamics [85]
Glide Systematic search [86]
GOLD GA [87]
PRO_LEADS Tabu search [88]
MOE-Dock Tabu search or simulated annealing [36]
FRED Systematic search, conformational ligand ensembles [89]
FLOG Systematic search [90]
FRED, Fast rigid exhaustive docking; FLOG, flexible ligands oriented on grid.
Virtual Screening 4027

both the relative positions of two molecules and their

conformations by a uniform set of internal variables
and uses a precalculated grid of interaction energy
values to speed up the computation.
GAs form a class of computational problem solving
methods adapting the main principles of biological
competition and population dynamics. GAs are
stochastic optimization methods; the problem is
encoded generally in the language of genetics. Initially,
a random population is generated that composes a set
of the potential solutions for a given problem. The
members of the population are represented by chromo-
somes; the genes correspond to certain variables. When
docking into protein structures, the variables for trans-
lation, rotation, and torsion angles are encoded in the
chromosome. Then genetic operators (crossing over
and mutation) are applied for the generation of new
Fig. 14 The schematic representation of the search tree.
Red nodes indicate energetically unfavorable conformations populations. The members of the new population are
during the ligand building. decoded and the estimation of the binding free energy
is evaluated. The generation of new populations is
terminated when a fixed number of population gener-
ation or a given score is reached. A prominent example
for the GA using docking algorithms is the genetic
Another method of systematic search is the use of optimization for ligand docking (GOLD) program.
databases of pregenerated conformations. Conformations Its special characteristic is the direct encoding of
are calculated once and the search problem is reduced H-bonding motifs. The latest version of AutoDock
to a rigid body docking procedure. This is the main uses a Lamarckian GA, which is a combination of a
concept of FLOG docking program.[91] traditional GA with local search method to perform
energy minimization.
Besides MC and GA methods, further heuristic
Random or stochastic methods algorithms have been developed for the docking prob-
(MC, GA, and tabu search) lem. Tabu search was found to perform well and
provided a straightforward searching strategy of the
The most widely used random approaches are the MC, PRO_LEADS program. This method operates on
genetic, and tabu search algorithms. MC algorithms randomly generated positions and these positions are
generate initial random conformations of a ligand in examined on the basis of the tabu list. This latter list
the binding cavity then calculate scores for these initial contains pregenerated solutions and provides restric-
configurations. The generation and scoring of config- tion on the search process. A random move of the
urations are repeated until the Metropolis criterion, ligand is ‘‘tabu’’ if it generates a solution that is not
which is used to determine whether the new confi- sufficiently different from the stored ones, unless its
guration is retained. According to the Metropolis energy is more favorable, therefore, the revision of
criterion, the new solution is accepted if the score of the search space is avoided.

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the new configuration (Enew) is better than the older

Virtual
one’s (Eold). If the configuration is not a new mini-
mum, it has to pass the Boltzmann-based probability Simulation methods (molecular dynamics
function test, otherwise the solution is rejected. and energy minimization)

Molecular dynamics simulation methods are currently

 

ðEnew
Eold Þ the most popular approaches. This method is for the
P ¼ exp analysis of protein flexibility and dynamic properties
kT
of molecular systems. With respect to docking, these
P
probability for the acceptance of the
simulations could provide a realistic view of the dock-
new configuration ing process, however, these calculations are still out
of reach. Therefore, dynamical simulations during
Internal coordinated mechanics (ICM) uses global the docking process are limited to the protein-ligand
MC minimization docking procedure,[83] describes complexes. Molecular dynamical simulations are often
 4028 Virtual Screening

unable to cross high-energy barriers as well, so an some kind of experimental data. Consequently, the
attempt is generally made to simulate different parts functions reflect the accuracy of these measurements.
of a complex at different temperatures or molecular Molecular size can influence the scores as well, gener-
dynamical simulations can be started from different ally the larger the molecule the better the score is,
starting ligand positions. In contrast to molecular however biological measurements do not support this
dynamical studies, energy minimization methods observation. As scoring functions are derived from
are rarely used as search techniques but often comp- X-ray structures, only the favorable interactions are
lement other methods such as MC methods. DOCK rewarded but unfavorable interactions are not pena-
performs an energy minimization step after fragment lized because information from the crystal structures
addition, followed by a final minimization step before cannot be obtained. Uncertainties in the protonation
scoring. states and the involvement of water in ligand binding
further complicate scoring. Some of the failures can
be corrected using consensus-scoring schemes, when
Scoring Functions the disadvantages and advantages of the various
scores might provide a combination, which is able to
The interaction between protein and ligand is a describe the main characteristics of the protein-ligand
reversible equilibrium reaction. The quantity to char- binding.
acterize this reaction is the free energy of binding.
Binding free energy values with reasonable accuracy
can only be calculated requiring large computational Force field-based scoring functions
resources. For scoring thousands of ligands, functions
to estimate the binding free energy must be computed The force field is a function expressing the energy of a
fast and without much computational power. Gener- system as a sum of e.g., diverse molecular mechanics
ally used scoring functions (Table 7)[92] estimate terms. Non-bonded energy terms of molecular mech-
the binding free energy for only one position/ anics force fields were first used to score protein-
conformation, and all of them share the assumption ligand complexes. The binding energy between a
that the full energy term can be decomposed into a ligand and a protein is most often estimated by the
sum of terms. summation of the electrostatic and van der Waals
However, the free energy of binding is a state func- energy terms. Standard force field scoring functions
tion so, in a strict physical sense, additivity is not have major limitations because they were developed
allowed. These scoring functions at low computational to model enthalpic gas phase contributions to struc-
cost are suitable for the calculation of an estimation of ture and energetics, and do not involve solvation
the binding free energy. Three main classes can be and entropic terms either. These scoring functions
separated: force field-based methods, empirical meth- are restrained by the fact that in VS, significant steric
ods, and knowledge-based methods. clashes cannot be avoided because the energy terms
All fast scoring functions share several deficiencies. have steep form at short interatomic distances (12-6
First of all most of them are fitted to or derived from Lennard–Jones potential; Dock score), the Lennard–
Jones potentials used for modeling van der Waals
interactions can lead to very strong repulsion, there-
fore, these terms are often scaled down (8-4
Table 7 Overview of frequently used scoring functions
Lennard–Jones potential; GOLD score). Recently
Type Scoring functions References developed scoring functions contain entropy terms
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Force field-based CHARMM [93,94] to increase the potential of specific molecular recog-
Virtual

methods AMBER nition, H-bonding are often designed in different

LIE ways. Below, two generally applied scoring functions
OWFEG are detailed in terms of use.
MM PB/SA The GOLD score (G-score) is based on the work of
GOLD/Gold(G) Jones et al. and concentrates on H-bonding inter-
DOCK/Dock(D)
actions. The H-bonding term contains desolvation of
Empirical methods PLP [80,95,96] donors and acceptors. A pairwise dispersion term
FlexX involves the contribution of hydrophobic interactions
ChemScore
and a molecular mechanical equation is used to calcu-
ScreenScore
HINT
late the internal energy of the ligand. Therefore, this
scoring function performs well when polar interactions
Knowledge-based DrugScore [97–99]
are dominant and has difficulty with ligands that are
methods PMF
primarily non-polar in nature.
Virtual Screening 4029

GOLD score: The scoring function implemented in DOCK

Version 4.0:
Protein-ligand:
" !
X X Aij Bij Protein-ligand:
EvdW þ EH-bond ¼
4
protein ligand
d8ij dij EvdW þ Eelectrostatic
# " ! #
X X Aij Bij qi qj
þ ðEda þ Eww Þ
ðEdw þ Eaw Þ ¼
b þ 332  
protein ligand
daij dij e dij dij
Internal ligand:
!
X Cij Dij Empirical scoring functions
EvdW þ Etorsion ¼
6
ligand
d12
ij dij
Empirical type functions form the second group of the
X 1  
n scoring functions. The binding energy is approximated
þ V 1 þ cosðjnjoÞ by a weighted sum of explicit hydrogen bonding and
ligand
2 jnj
hydrophobic contact terms. Many additional terms
and different functional forms are used for the scoring.
where Aij, Bij, Cij, and Dij are van der Waals param-
Some functions describe H-bonds with, e.g., distance
eters for atoms i and j, and dij is the interatomic
terms while others penalize angular deviation from
distance. Eda, Eww, Edw, Eaw are the donor–acceptor
the idealized values. ThePbinding free energy can be
individual energy values, V is the torsional barrier, jnj is
estimated as: DGbind ffi DGi fi, where fi corresponds
the periodicity, o the torsion angle.
to a weighting factor and DGi represents an interaction
GOLD score is the original GOLD scoring function
term. The weighting factors are obtained from
implemented in GOLD docking program package. An
regression analysis using experimentally determined
optional H-bonding energy term can be added.
binding energies and 3-D structural information. The
appeal of empirical scoring functions is that their terms
Protein-ligand:
can be simple to evaluate but they are based on
EvdW þ Eelectrostatic approximations similar to force field functions. The
" ! # disadvantage of this class of scoring functions is the
X X Aij Bij qi qj
¼
b þ 332   dependence on the dataset used for the regression
protein ligand
daij dij e dij dij analysis and fitting. Empirical scores contain a rotor
term, which estimates entropy penalties on binding
Internal ligand: from a weighted sum of the number of RBs in ligands.
Currently used terms involve an incomplete descrip-
EvdW þ Eelectrostatic tion of solvation/desolvation energy upon ligand
" !#
X Aij Bij qi qj binding. In the next paragraph, three of the most
¼ a
b þ 332   commonly used scoring functions are discussed in
dij dij e dij dij 
ligand terms of their use.
þ EH-bondðoptionalÞ LUDI separates the H-bonding term into neutral
and ionic H-bonds and calculates the hydrophobic
where qi and qj are atomic partial charges and e(dij) is contributions based on the molecular surface area.

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a dielectric function. X

Virtual
Dock score (D-score) considers both electrostatic DGbind ¼ DGH-bond f ðDR; DaÞ
and hydrophobic contributions to the binding energy H-bond
X
but entropic terms. A distance dependent dielectric þ DGionic f ðDR; DaÞ
attenuates polar interactions. ionic
X  
þ DGhydrophobic Ahydrophobic 
Protein-ligand: hydrophobic

EvdW þ Eelectrostatic þ DGrotor Nrotor þ DG0

" ! #
X X Aij Bij qi qj where Ahydrophobic is the molecular surface area, DR is
¼
6 þ 332
protein ligand
d12
ij dij eðdij Þdij the deviation from the ideal H-bond length, Da is the
  deviation from the ideal H-bond angle, DG0 is a
e dij
a distance-dependent dielectric function regression constant.
 4030 Virtual Screening

FlexX score is based on Böhm’s work.[80] This equa- models in the context of protein structure predictions.
tion is basically the DG for an ideal H-bond, ionic, To establish a function for scoring, the i, j, and k
aromatic, etc., interaction, adjusted by a penalty variables are assigned to protein and ligand atom
function that depends on the deviation from the ideal types and their interatomic distances. The occurrence
geometric values of two interacting elements. frequency of their contacts is a measure of the energetic
X contribution to the binding. DrugScore and potential of
DGbind ¼ DGH-bond f ðDR; DaÞ mean force (PMF) are popular implementation of this
H-bond type of scoring functions:
X DrugScore:
þ DGionic f ðDR; DaÞ
ionic X X
X DW ¼ g DWij ðr Þ þ ð1
gÞ
þ DGaromatic f ðDR; DaÞ protein ligand
aromatic
X " #
X X
þ DGcontact f ðDRÞ  DWi ðSAS; SAS0 Þ þ DWj ðSAS; SAS0 Þ
contact
ligand protein
þ DGrotor Nrotor þ DG0
SAS
solvent accessible surface area
ChemScore is based on a diverse training set of Wij
distance-dependent pairwise potential
82 receptor-ligand complexes. It uses four terms that
estimate contributions to binding energy from lipo- g
adjustable weight factor
philic interactions, metal-ligand binding, hydrogen
bonding, and loss of ligand flexibility. ChemScore is Parametrized pairwise potential PMF score:
more accurate and robust than many other widely used
X X  
functions. PMF ¼ Aij dij
X protein ligand
DGbind ¼ DGH-bond f ðDR; DaÞ " #
H-bond   j rijseg ðr Þ
Aij dij ¼
kB T ln fvolume corr ðr Þ
X rijbulk
þ DGmetal f ðDR; DaÞ
metal kB
Boltzmann constant
X j
þ DGlipo f ðDRÞ fvolume corr ðr Þ
ligand volume correction factor
lipo

X  rijseg(r) – the number density of a ligand-protein atom

0

þ DGrotor f Pnl ; Pnl þ DG0 pa of type ij at a certain atom pair distance r
rotor
rijbulk – the number density of a ligand-protein atom pair
0 of type ij in a reference sphere with a radius of 12 Å
where Pnl and Pnl are the fraction of non-lipophilic
atoms on either side of the frozen bond.
Postprocessing
Knowledge-based scoring functions
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Filters such as the ones used for preparing the input

Virtual

In knowledge-based functions, the ‘‘knowledge’’ that screening databases can also be applied here. Although
is implicitly encoded in the protein-ligand complexes is after docking, this step does not save time, e.g.,
tried to be captured. These scores are based on the con- compounds not suitable for further drug development
cept of the inverse formulation of the Boltzmann law: can be filtered out. Currently used scoring functions are
inadequate for the precise binding affinity predictions.
Eij ¼
kT lnðpijk Þ þ kT lnðZÞ Therefore, additional postprocessing strategies are
Eij
potential of mean force often advised to apply for the minimization of the
number of false positives in the ranked database
pijk
probability density and to propagate the true hits to the top of the list.
Z
partition function A popular strategy is the consensus scoring. In this
approach, scoring functions first can be separately
This approach has been applied to assemble potentials applied for selection of the best docked position and
from databases of protein structures to score protein then for ranking the best positions. The top ranking
Virtual Screening 4031

molecules then can be rescored by other scoring

functions, and can be ranked again. Scoring functions
can be weighted, so a linear combination of the scores
can also be applied. An appropriate scoring scheme
for the particular protein target can be worked out.
Several variety of consensus scoring approaches have
been frequently applied to increase the performance
of VS.[100]
Besides the consensus scoring and 1-D filters (MW,
log P etc.) can decrease the artificial enrichment,[101]
which occurs as larger molecules are usually scored
better. Thereby normalized scores can be calculated
with e.g., solvent accessible surface, MW etc., so as to
compensate the large molecular surface.
From the visual inspection of the docking solutions
to the involvement of solvation or entropic effects,
scores are often corrected to improve the prediction
of the binding free energy and as a direct consequence
the enrichment of active compounds. Fig. 16 Crystal structures of CDK-2 (red) and ChK-1
(blue), both bound with staurosporin inhibitor and aligned
by homology.
Case Study: VS for Checkpoint
Kinase-1 Inhibitors using
Knowledge-Based VS
inhibitors for kinase targets such as cyclin-dependent
Kinase inhibitors have recently become a major area of kinase,[103] and epidermal growth factor receptor
drug discovery and structure-based design.[102] Novel kinase[104] have recently been discovered. As many
3-D structures are publicly available for kinases, they
seem to be an ideal target for VS studies. Although
only a few case studies have been published in the
pharmaceutical industry, VS is often performed for

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Virtual

Fig. 17 The ATP binding site of CDK-2 containing stauros-

Fig. 15 Screening cascade taken to the virtual screen for porin inhibitor. The polar contacts between the ligand and
ChK-1 inhibitory activity. protein are seen in black.
 4032 Virtual Screening

checkpoint kinase-1 (ChK-1). Their strategy exploits

the chemical and structural information available for
kinase inhibitors. Compounds with a minimal kinase-
binding motif were docked; only then the docked poses
were rescored using a scheme customized for a kinase,
the CDK-2. High scoring compound were further
investigated visually and selected for biological testing.
Thirty-six effective inhibitors out of the 103 best rank-
ing compounds were found.
The overview of the screening cascade is shown in
Fig. 15. The screening library was first prepared for
docking. The preprocession of this database involved
a filtering step of molecules having MW greater than
600 Da, and having rotatable bonds more than 10.
Large, non-drug-like, and flexible molecules were
excluded. Flexible molecules are often docked
improperly using the FlexX algorithm. Tautomeric
and protonation states of the ligands were generated
using Leatherface.[105] 3-D coordinates were computed
Fig. 18 The ATP binding site of ChK-1 containing stauro- using Corina.[106] Compounds without an appropriate
sporin inhibitor. The polar contacts are between the ligand kinase-binding motif were filtered out by using
and protein seen in black. Plurality (H-bond donors/acceptors).
The ChK-1 crystal structure (Fig. 16) was proto-
nated at crystallization pH. Active site contained all
the residues within 6.5 Å of the bound inhibitor. Essen-
kinase targets. As kinases share a conserved ATP bind- tial pharmacophore constraints for the Cys87 (back-
ing site, the discovery of truly novel inhibitors can be bone NH) and Glu85 (backbone CO) residues were
limited. The existence of the conserved ATP binding given (Figs. 17 and 18).
sites does suggest that a general VS strategy can be CDK-2 was chosen as a surrogate for ChK-1
employed in searching for kinase inhibitors. because more activity data were available for the
Lyne et al.[105] proved an evidence for the above CDK-2 that time. An enrichment study using a screen-
assumption using knowledge-based strategy for ing database of 8000 inactive and 100 of active
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Virtual

Fig. 19 Enrichment plot for CDK-2 using all the possible scoring schemes. P, PMF; D, Dockscore; C, Chemscore; F, FlexX-
score; G, Goldscore. (Copyright is permitted by the Copyright Office of the American Chemical Society).
Virtual Screening 4033

all the raw scores. For CDK-2 PMF and FlexX, scoring
functions were found to give the best enrichment factors
(EF). Thus this scoring scheme was applied for ranking
the docked solutions for ChK-1. The enrichment plot is
shown in Fig. 19. Enrichment factors (EFs) assess the
quality of the rankings:

 
Nactiveð%Þ =Nð%Þ
EFð%Þ ¼
Nactive =NðallÞ

Fig. 20 Two examples of the hits found by virtual screening

for ChK-1.
where EF(%) is given at the percentage of the ranked
database, Nactive(%) is the number of active compounds
in a selected subset of the ranked database, N(%) is the
compounds was performed. Three hundred docked number of compounds in the subset, and Nactive and
poses per molecule were saved. For rescoring, the Cscore Nall are the number of active molecules and the number
module of SYBYL was used. Z-scores were calculated of compounds in the screening database. EF were not
for all the possible scoring functions combinations: scaled, i.e. absolute values depend on the ratio of active
and inactive molecules and should be compared to the
X xi
x

maximum (ideal) achievable EFs.
Z ¼ This study proves to be a successful case study in
i
s
the field of protein kinase inhibition with ATP com-
where x is the raw score for a pose, x

average raw score petitive compounds (Fig. 20). The hit rate was 36%,
for all the poses, and s is the standard deviation for and was achieved by screening 103 compounds.

Table 8 Summary of recently published structure-based virtual screening studies indicating EFs
Short description

Target protein Program Screening database Actives EF(%) Ideal EF(%) EF/Ideal EF Reference
Thymidine kinase DOCK ACD: 990; (filters: Mw, 10 7(10) 10 0.7 [107]
FlexX reagents, inorganic), 8(10) 10 0.8
random set from
Estrogen receptor a GOLD 10 10(10) 10 1
the filtered database,
Gasteiger-Marsili charges
Protein tyrosine DOCK ACD: 179805; (filters: 1327 7(10) 10 0.7 [108]
phosphatase-1B FlexX metals, isotope, clog P), 9(10) 10 0.9
Glide random set from the 9(10) 10 0.9
filtered database,
Protein kinase B DOCK 266 0.1(10) 10 0.01

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Gasteiger-Marsili

Virtual
FlexX charges, ionization 1(10) 10 0.1
Glide 5(10) 10 0.5

Cyclooxygenase-2 FRED WDI: 7528; (filters: Mw, 128 5(10) 10 0.5 [109]
FlexX similarity, rotatable bonds, 6.2(10) 10 0.62
Glide macrocycles removal, 6.3(10) 10 0.63
steroid-type scaffolds,
Estrogen receptor a FRED pharmacophore points, 55 7.9(10) 10 0.79
FlexX ionization at pH 7) 5.8(10) 10 0.58
Glide 7.5(10) 10 0.75
p38 kinase FRED 72 1.9(10) 10 0.19
(Continued)
 4034 Virtual Screening

Table 8 Summary of recently published structure-based virtual screening studies indicating EFs (Continued)
Short description

Target protein Program Screening database Actives EF(%) Ideal EF(%) EF/Ideal EF Reference

FlexX 7.9(10) 10 0.79

Glide 4.1(10) 10 0.41
Gyrase B FRED WDI: 7528; (filters: 36 4.1(10) 10 0.41 [109]
Mw, similarity,
rotatable bonds,
macrocycles removal,
steroid-type scaffolds,
pharmacophore points,
ionization at pH 7)
FlexX 7(10) 10 0.7
Glide 5.8(10) 10 0.58
Thrombin FRED 67 7.9(10) 10 0.79
FlexX 8.1(10) 10 0.81
Glide 8.3(10) 10 0.83
Gelatinase A FRED 43 4.1(10) 10 0.41
FlexX 6.3(10) 10 0.63
Glide 6.1(10) 10 0.61
Neuraminidase FRED 51 8.3(10) 10 0.83
FlexX 6.2(10) 10 0.62
Glide 9.8(10) 10 0.98
Cyclooxygenase-2 FlexX 128 20(1) 59 0.34
Estrogen receptor a FlexX 55 39(1) 100 0.39
p38 kinas FlexX 72 18(1) 100 0.18
Gyrase B FlexX 36 21(1) 100 0.21 [96]
Thrombin FlexX 67 25(1) 100 0.25
Gelatinase A FlexX 43 16(1) 100 0.16
Neuraminidase FlexX 51 22(1) 100 0.22

Neuraminidase GOLD ATLAS 15 43(1) 100 0.43 [101]

(in house database):
3.1 million
PTP1B 25 20(1) 100 0.2
CDK-2 Mw > 250 41 10(1) 100 0.1
CDK-2 Mw < 250 23 23(1) 100 0.23
Estrogen Receptor 20 60(1) 100 0.6
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Virtual

a (agonist)
Estrogen Receptor 17 58(1) 100 0.58
a (antagonist)
PTP1B, Protein tyrosine phosphatase-1B. The table is provided to allow comparison of docking schemes with respect to the algorithms and target
proteins.

Hits corresponded to four chemical classes. These are Success Stories

the results for this particular study only and it is not
premised that these could be observed in general with Improvements in docking algorithms and the increasing
VS for protein kinases. However, this study gives a number of scoring functions have resulted in a large
proof of the applicability of this strategy in other VS number of successful VS studies. In this section, a collec-
study performed for protein kinases. tion of successful screening stories (Table 8) is described.
Virtual Screening 4035

CONCLUSIONS ARTICLES OF FURTHER INTEREST

Besides experimental high-throughput screening, VS Computer-Assisted Drug Design, p. 714.

has emerged as a knowledge-based alternative that Drug Design: Basic Principles and Applications,
can be applied in drug discovery. Recently published p. 1362.
studies have revealed that the two approaches are
rather complementary than competitive.[110] Owing to
the involvement of ever increasing numbers of com- REFERENCES
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Virtual
Water for Pharmaceuticals
Leonid Shnayder
Aker Kvaerner Pharmaceuticals, Bridgewater, New Jersey, U.S.A.

INTRODUCTION the steam system). On the other hand, water used as an

ingredient for an injectable drug has to meet much more
Water is one of the key utilities in most pharmaceutical stringent quality requirements. The US Pharmacopeia
facilities. It is used as a solvent, product ingredient, (USP) contains specifications for several grades of water
cleaning agent, and for many other applications. Some used in the preparation of medicinal products. The two
of those applications require water of higher purity grades most often used in the pharmaceutical plants are
than typically found in municipal water supplies. USP Purified Water and Water For Injection (WFI). As
Therefore companies install private water purification the name implies, WFI is used for the preparation of
systems that become a critical part of their utility injectable drugs, whereas USP Purified Water can be
infrastructure. used in the manufacturing of tablets, capsules, creams,
Why does the potable water from municipal supply lotions, etc. These types of water are called ‘‘compen-
require further treatment for pharmaceutical man- dial’’ because their quality is specified in an official
ufacturing? To understand this, let us look into the nationally recognized standard such as USP. In addi-
two main water sources for municipal plants: ground- tion, many companies use various non-compendial
water aquifers (well water) and surface supplies (rivers water systems designed for specific needs. While design-
and lakes). As rainwater falls onto the Earth’s surface, ing the facility, it is important to decide on the proper
it picks up and dissolves atmospherical gases (oxygen grades of water to use in various applications. Compen-
and carbon dioxide), industrial emissions such as nitric dial waters are typically very expensive, not only
and sulfuric oxides, as well as bacteria and dust parti- because of required treatment steps, but also because
cles. As is passes through the ground into the aquifers, of extensive validation and testing requirements. There-
the water is filtered and becomes quite clean, free of fore it is prudent to limit their use to processes where
most particles and organic contaminants. However, it such water becomes an ingredient of the pharmaceutical
now dissolves minerals contained in the limestone product, is in direct contact with such product, or is
and other geological materials. That adds calcium, used for the final rinse of critical process equipment.
magnesium, sodium, iron, chlorides, sulfates, and other For most other applications, various non-compendial
ions to the ground water. The surface waters found in grades of water (potable, softened, deionized, etc.) can
the lakes and rivers collect organic materials and par- be successfully used without any regulatory conflicts.
ticles from vegetation and ground runoff. It is not uncommon to install a water system that pro-
Municipal water plants provide limited treatment, duces water that could easily meet the USP specification
mostly intended to make the water safe to drink. A yet is not designated as USP Purified Water to avoid
lot of contaminants, such as salts, dissolved gases, unnecessary expenses for regulatory compliance.
and organic materials contained in natural sources, The current edition of USP[1] establishes the follow-
remain in the municipal water supply. In addition, ing requirements for USP Purified Water:
chlorine or other disinfectants are often added as part
of the treatment process to control microbial contami-
Purified Water is prepared from water complying
nation. For many critical applications required in with the federal regulations for drinking water
pharmaceutical plants, such water quality is not suffi-
Purified Water contains no added substance
cient, and further treatment is necessary.
Purified Water is obtained by a suitable process

Conductivity does not exceed set level (1.3 mS/cm
at 25 C)
Water–Zeta

GRADES OF WATER USED IN THE
Total organic carbon (TOC) does not exceed set
PHARMACEUTICAL FACILITIES level (500 ppb)

The water purity required varies widely with the appli- The requirements for WFI are as follows:
cation. For example, water used to feed the boilers pro-
ducing plant steam only needs to be softened (to avoid
Meets all the requirements for ‘‘Purified Water’’
scale buildup) and deaerated (to avoid corrosion in
Is obtained by distillation or reverse osmosis (RO)
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-120018249
Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved. 4039
 4040 Water for Pharmaceuticals

Contains not more than 0.25 USP endotoxin units The most common process used for this is depth
per milliliter filtration through a bed of sand or similar material

Is prepared using suitable means to minimize charged in a vertical vessel. The incoming water flows
microbial growth from top to bottom. To improve the efficiency of such
filters, two or more layers of media with various particle
The USP does not specify allowable concentrations of sizes are used. Coarse and less dense material such as
microorganisms in the official monograph, but recom- anthracite is located at the top of the bed, whereas finer
mends action limits of 100 colony-forming units per ml and denser particles of sand are placed at the bottom.
(CFU/ml) for Purified Water and 0.1 CFU/ml for WFI. Such multimedia filters can remove most particles
Putting aside the items related to the source water larger than 10–20 mm. Periodically the filter bed is back-
and treatment procedures, we can summarize the require- washed by reversing the flow direction (from bottom to
ments for Purified Water itself as follows: top) and by increasing the flow rate. During backwash,
the captured particles are removed and sent to drain,
1. Low conductivity (high resistivity). This test is whereas heavier particles of the filter bed remain in
intended to show that water contains a minimal the vessel and settle back at the end of the cycle.
amount of ions such as calcium, magnesium, In smaller water purification systems, we can often
sodium, iron, chloride, sulfate, etc. The inherent find a cartridge-type depth filter used for the particle
presence of hydrogen and hydroxide ions deter- removal. The cartridge is made of cellulose, cotton,
mines the theoretical limit of Purified Water or polymer fibers, which trap dirt particles throughout
conductivity: approximately 0.05 mS/cm (resis- its volume. Once a cartridge reaches its holding
tivity 18 MO cm) at a pH of 7.0. The practical capacity, it is disposed of and replaced by a new one.
limits specified in the USP are in the range of
1–5 mS/cm, depending on pH and temperature.
Adsorption of Dissolved Organics and Chlorine
2. Low TOC (less than 500 ppb). Water shall con-
tain a minimal amount of organic compounds.
Activated carbon filters often found in pharmaceutical
Such compounds are undesirable for two main
water purification systems are used to remove chlorine
reasons: they may be toxic, and/or they may
and many dissolved organic materials found in the
serve as sources of nutrition for microorganisms.
incoming potable water. Physically, a carbon filter
3. Low microbial count. Water shall contain a
looks exactly like the multimedia filter: a vertical tank
minimal amount of viable microorganisms,
with a valve manifold next to it. The main difference is
including spores.
that the filter bed consists of activated carbon particles
4. Low endotoxin level (required for WFI only). The
with a very high surface area. These can absorb and
term ‘‘endotoxin’’ applies to organic compounds
retain amazing amounts of organic contaminants and
that cause harmful effects when injected in the
chlorine. The major disadvantage of carbon filters is
bloodstream of laboratory animals. Such com-
that they may become a breeding ground for bacteria
pounds are lipopolysaccharides produced as a
thriving in an environment without chlorine (which is
result of microbial growth or microbial destruction.
typically adsorbed in the upper part of the bed) and
with plenty of nutrients. Therefore carbon filters are
usually periodically sanitized with hot water to contain
WATER PURIFICATION SYSTEMS microbial growth. The bed is also backwashed fre-
quently to remove captured particles. However, activated
As naturally occurring water has a variety of contami- carbon cannot be economically regenerated in the field;
nants, there are many treatment processes developed to once it has reached its adsorption capacity (typically
remove those contaminants. A typical pharmaceutical after 1–5 years), the bed is replaced with fresh carbon.
water purification system contains several unit opera- It is recommended to use acid-washed activated carbon
tions designed to remove various components. for water purification because the minerals resident in
the charcoal may lead to leaching of the metal oxides,
increasing the water hardness.[2]
Particle Removal
Water–Zeta

The municipal water treatment plants use a process Ion Exchange

called ‘‘clarification’’ based on gravity settling to
remove a majority of the large particles suspended in An ion exchange process is based on the ability of cer-
water. However, some of the suspended materials tain synthetic resins to selectively adsorb either cations
remain in the potable water and need to be removed or anions, and to release (exchange) other ions based
as part of the pharmaceutical water purification. on their relative activity. The water passes through a
Water for Pharmaceuticals 4041

bed of resin beads in a vertical vessel (again, looking Membrane Filtration

very similar to a multimedia filter), where such
exchange takes place. Once all the active sites on the Unlike depth filters where particles are captured
resin bed are filled with the new ions, the resin needs throughout the filter volume, membrane filters are
to be regenerated by passing an appropriate chemical surface-type filters: they stop particles larger than the
solution through the bed. pore size right at the upstream surface of the poly-
The most common type of ion exchange system is a meric membrane. Naturally, such filters have very
water softener. Its main function is to remove calcium limited holding capacity: once most of the surface
and magnesium ions from hard water. These are the holes are plugged with a single particle each, the fil-
components that cause the formation of the hard-to- tration process stops. To overcome this inherent limi-
remove scale in tea kettles; they have a similar effect tation, engineers invented a process called ‘‘crossflow’’
on boilers, distillation stills, and membrane filtration or ‘‘tangential flow’’ filtration. In such a system
equipment. The polystyrene resin has a greater affinity (Fig. 1), fluid moves at high velocity parallel to the
for multivalent ions such as calcium and magnesium membrane, with only a small fraction of the feed flow
than for monovalent sodium. When the water passes penetrating through it because of a pressure differen-
through the bed, calcium and magnesium ions are tial. The particles or molecules that could not penetrate
captured and exchanged for the sodium adsorbed on through the membrane are swept by the parallel flow
the fresh resin. Once the resin is exhausted, it is and exit the system in a stream called ‘‘concentrate’’
regenerated by a 10% solution of sodium chloride. because it contains a higher concentration of particles
The high concentration of sodium ions in such a than the feed stream. The stream penetrating through
solution shifts the equilibrium of the adsorption– the membrane is called ‘‘permeate.’’ Because of the
desorption process, resulting in the displacement of ‘‘sweeping’’ action of the crossflow stream, membrane
captured multivalent ions with sodium ions. The holes do not plug up and such systems can function for
regeneration cycle can be repeated many times, making a long time between cleanings. Depending on the pore
softener resin last for years. Softening is a very simple size, membrane filtration processes are called microfil-
and inexpensive process requiring only salt for resin tration, ultrafiltration, and reverse ozmosis (RO).
regeneration, and it is widely used not only in high- Microfilters use membranes with pores in the 0.1–
purity water systems but also in many utility water 1 mm range. They can filter out particles of dust, acti-
systems. vated carbon, and ion exchange resin fines, and most
A similar process also takes place in ion exchange microorganisms. Microfilters require low differential
deionizers, designed to remove practically all foreign pressures (5–20 psi) and are available both as normal
ions from water. Deionizers use two types of resin: flow (‘‘dead end’’) and crossflow configurations. In
one exchanges positively charged ions (cations) such pharmaceutical water purification systems, they are
as sodium, calcium, and magnesium with hydrogen often used as disposable cartridge filters after activated
ions (Hþ); another exchanges negative ions (anions) carbon filters, softeners, and ion exchange beds.
such as chloride, sulfate, and bicarbonate with hydrox- Ultrafilters use membranes that reject not only solid
ide (OH). Although the ion exchange process can particles but also dissolved matter with high molecular
produce very-high-quality water, its major drawback weight. The ‘‘molecular weight cutoff’’ point of such
is the type of chemicals required for resin regeneration: membranes varies in the range of 10,000–100,000 Da.
cation resins are regenerated by strong acid, and anion They can remove bacteria, endotoxins, colloidal con-
resins are regenerated by strong base solutions. taminants, and large organic molecules. Ultrafilters
Another drawback important for the pharmaceutical typically require differential pressure between 10 and
water systems is the fact that ion exchange resins can 50 psi, and they are also available in both normal and
become a breeding ground for microorganisms. For crossflow configurations. In water purification systems,
smaller systems, it is common to use service-type ion ultrafilters are sometimes used as a polishing step after
exchange units, where the vessel with exhausted resin ion exchange units.
is taken offsite for regeneration, whereas larger systems
have to be regenerated on site, resulting in the need to
handle concentrated acids and bases and to neutralize Feed Concentrated
Water–Zeta

waste solutions. A relatively new process gaining popu-

Membrane
larity in the field is the continuous deionization system,
which represents a combination of ion exchange and
Permeate

membrane separation technologies. It uses electrical

current to continuously regenerate the ion exchange
resin simultaneously with the water treatment process,
eliminating the need to handle strong chemicals. Fig. 1 Tangential flow filtration.
 4042 Water for Pharmaceuticals

Interconnector
Permeate tube

O-Ring Concentrate
flow
Concentrate
Feed flow flow
Mesh spacer
Permeate flow

Feed flow

Membrane

Mesh spacer Permeate

Adhesive carrier
bond
(at edges of membrane envelope)

Fig. 2 Spiral-wound membrane element. (Courtesy of Osmonics, Inc.)

Reverse osmosis is a filtration process using a mem- while expensive, allows the removal of almost all of
brane with the pore size in the range of 1–10 Å. Such the organic and inorganic impurities and achieves
membranes reject not only most organic compounds, very-high-quality water. It is also considered the
bacteria, and viruses, but also 90–99% of all ions. safest way to avoid microbial and endotoxin conta-
Because of the extremely small pore size, RO systems mination. That is why distillation remains the domi-
operate at high differential pressures—typically from nant method of producing WFI. Feed water to the
100 to 500 psi. Practically all commercial RO systems distillation stills is often pretreated with RO and/or
are designed in the crossflow configuration. The most other methods to reduce scaling and to increase
common physical membrane configuration for RO distillate quality.
systems uses spiral-wound elements (Fig. 2) placed One of the possible configurations for the pharma-
into horizontal pressure vessels (essentially pipes). In ceutical water purification system is shown in Fig. 3.
the production of the pharmaceutical Purified Water, The first three stages in that system (multimedia fil-
it is very common to use double-pass RO systems with tration, softening, and carbon filtration) are considered
two filtration stages connected in series. Such systems pretreatment steps for the RO unit that conducts final
can easily produce water meeting the requirements purification. The cartridge filter shown before the RO
for USP Purified Water and even WFI.[3] However, is provided to protect the membranes from the fine
European regulations do not allow RO to be used as carbon particles. There are many variations of the
a final treatment step for the production of WFI. treatment systems: sometimes carbon filter is placed
This also affects U.S. facilities as the common practice before a softener, sometimes it is eliminated completely,
in the United States is to build facilities compliant
with both U.S. and European regulations. In the last
10–15 years, RO has become the most common way
to produce pharmaceutical Purified Water—either as
a final treatment step, or as pretreatment for the City water

distillation stills.
Multi-media filter Water softener Carbon filter

Distillation

Distillation is a process that involves the evaporation

Water–Zeta

of water and then the condensation of the resulting Purified water Two-pass RO
steam. Most dissolved contaminants do not evaporate to storage tank system
at the temperature of boiling water, and therefore Cartridge filter
do not pass into the steam and condensate streams. Concentrate to drain

To improve energy efficiency, distillation is usually con-

ducted in multiple-effect stills designed to recover most
of the energy spent on evaporating water. Distillation, Fig. 3 Pharmaceutical water purification system.
Water for Pharmaceuticals 4043

and chlorine removal is accomplished by adding reduc- membranes that constitute endotoxins, both during
ing chemicals such as sodium sulfite; often ultraviolet their normal growth and after the cell’s destruction.
(UV) lights are added to improve microbial control That is why elimination of the Gram-negative micro-
and/or to reduce TOC levels, etc. The systems organisms from the water systems is one of the primary
designed to produce WFI most often include distil- goals. That also explains why test samples taken from
lation still as a final treatment step. the water system soon after sanitization sometimes
show higher levels of endotoxins that those observed
before: successful sanitization kills all or most of the
STORAGE AND DISTRIBUTION SYSTEMS bacteria in the biofilm, and pieces of their membranes
FOR PHARMACEUTICAL WATER are released in the bulk water stream where they
can be detected as endotoxins. Another consequence of
Further discussion assumes that water leaving the puri- bacterial growth in biofilms that must be kept in mind
fication system and entering the storage tank meets is that results of microbial sampling of the water sys-
all of the above requirements either for USP Purified tems are only indirectly correlated with the actual situ-
Water or WFI. The goal when designing and operating ation. First, the sample is usually taken from the bulk
the storage and distribution system is to keep the water liquid stream whereas most bacteria reside on the pipe
at these purity levels, preventing any of the four param- and equipment surfaces. Second, the testing tech-
eters listed above from exceeding allowable limits dur- niques, including the type of growth media used to
ing storage. In particular, the engineers and operators cultivate the sample before counting colonies, can have
have to ensure that: a great impact on the test results. Therefore it is no
accident that USP does not specify an allowable con-

Water is protected against ionic and organic con- centration of microorganisms in the water samples as
tamination (that would lead to an increase in the a pass–fail criteria similar to conductivity or TOC,
conductivity and TOC respectively) but only as a helpful indirect indicator that shall be

The system is protected against physical entry of used as an alert or action limit in the efforts to main-
foreign particles and microorganisms tain microbial control.

Microbial growth is prevented or minimized It is important to point out that a small number of
microorganisms surviving in the purified water system
Before moving on to the description of the typical do not necessarily affect the water quality. Because of
storage and distribution systems and their compo- the lack of nutrients and other unfavorable conditions,
nents, it is beneficial to say a few words about the issue the microbial growth in such waters is very slow. That
of the microbial growth that presents probably the provides operators with an option to maintain such
biggest challenge and causes most concerns among systems in the state of microbial control by sanitizing
engineers and operating personnel. them periodically rather than continuously, as long
Microorganisms are ubiquitous; they can be found as the frequency is high enough to keep the bacterial
in a very wide variety of environments. High-purity counts at acceptable low levels.[7,8]
water systems are no exception. Although such water Let us consider some of the engineering practices
contains very few nutrients necessary to sustain life, used to achieve the above mentioned goals. For a more
some types of bacteria developed skills that allow them detailed description, refer to the ISPE Baseline Phar-
not only to survive but also to multiply in such waters. maceutical Engineering Guides[9] recently developed to
They do it for the most part by attaching themselves to help industry professionals in designing and operating
the pipe and other system’s surfaces and by creating a purified water systems.
thin film containing microorganisms, organic and
inorganic nutrients (obtained partially from the water
and partially from the dead bacteria), and products Overall System Schematics
of bacterial metabolism and decay.[4–6] Such film,
appropriately called biofilm, constitutes a niche envi- One of the typical flow diagrams for the pharmaceuti-
ronment that is very different from the bulk of the cal water storage and distribution system is shown in
water system’s volume, and may be reasonably com- Fig. 4. The system includes a vertical storage tank,
Water–Zeta

fortable for such bacteria. The majority of micro- circulation pump, sanitary heat exchanger, and piping
organisms capable of forming biofilms in the purified designed in the form of complete loop with point-of-
water systems are classified as Gram-negative organ- use valves located at the minimum distance to the loop.
isms in reference to their quality of not retaining a The pump continuously circulates the water from the
dye after staining (Cristian Gram was the inventor of tank via distribution piping and back to the tank.
the staining method). Such organisms have a tendency The water temperature in the system is maintained at
to shed lipopolysaccharidic materials from the cell 65–85 C to maintain microbial control; any organisms
 4044 Water for Pharmaceuticals

the main header from the ceiling space to a convenient

working level where a point-of-use valve would

Steam
Purified water Vent be easily accessible. That adds a lot of elbows and
from WFI still Heater associated pressure drop to the system. In cases
where the point-of-use valve is automated, this can

Cond
Storage sometimes be avoided by locating the valve at the
tank
80˚C header, away from the actual point of use. In such case,
the valve only needs to be accessible for maintenance,
but sanitization of the branch line may have to be
addressed.
The storage tank is usually equipped with a
Points of use
sterilizing-grade (0.2 mm) hydrophobic vent filter, and
sometimes also with a nitrogen blanketing system, to
prevent the introduction of particles and organisms
Circulation pump during normal inbreathing. The additional benefits of
the nitrogen blanket are the creation of oxygen-free
Fig. 4 Purified water storage and distribution system. atmosphere in the storage tank’s upper space (better
for microbial control and corrosion protection), and
the avoidance of the slight vacuum conditions during
that found their way into the system would be killed or the inbreathing via vent filter. The vent filter on the
at least prevented from multiplying at such tempera- storage tank is often heat-traced to stay hot, both for
tures. The heat exchanger is used to compensate for microbial control and for prevention of moisture
the heat losses occurring in the piping system. The condensation.
water is returned to the tank via one or more spray The water circulation pump must be of sanitary
balls, thus providing continuous flushing of the tank’s design and may be equipped either with a single or a
top head and upper walls, and keeping them at the double mechanical seal. In the latter case, the purified
high temperature. The entire system (tank, piping, water from the pump’s discharge is used as a seal flush
and other components) is insulated for heat conser- fluid (Fig. 5). If a spare circulation pump is installed
vation. To minimize the ‘‘dead legs’’ at the points of for reliability, it is important to ensure that it does
use, it is common to design the loop with U-shaped not constitute a long ‘‘dead leg.’’ This can be avoided
sections located at each such point, in essence bringing either by valving it off and draining it free of water
Water–Zeta

Fig. 5 Double mechanical seal with purified water flush arrangement. (Courtesy of Fristam Pumps, Inc.)
Water for Pharmaceuticals 4045

when not in use, or by allowing a small flow of hot

Steam
water to constantly circulate through the spare pump.
The circulation flow rate is usually selected based Purified water
from WFI still Vent
not only on the anticipated water usage rates, but also Heater

Cond
on the requirement to maintain certain minimum Storage
velocity (typically 5 ft/sec) during periods of no usage. tank
80˚C
The minimum velocity requirement comes from a
widely held belief that 5 ft/sec is necessary to create a Clean
steam
turbulent flow and that such flow prevents the accumu-
lation of the biofilm by mechanical shear forces. How- Points of use Chilled
water
ever, this notion is not supported by experimental return
results or analysis of the fluid dynamics.[10,11] First, Cooler
the flow becomes turbulent at velocities well below Circulation pump

5 ft/sec. Second, even with the turbulent flow, there is

always a thin film of liquid adjacent to the piping wall Points of use for Chilled
cooled water water
that exhibits laminar flow. The thickness of this film, supply
although dependent on the velocity, is certainly much
higher than the size of typical bacteria. However, it Fig. 6 Purified water storage and distribution system with
point-of-use cooler.
would be unfair to say that the circulation velocity
does not matter. It shall be selected not only based
on the pipeline hydraulics, but also considering its
effect on maintaining required water temperature at (typically 20–25 C). It is important to point out that
the various points in the system, including any ‘‘dead such system cannot be expected to maintain exact loop
legs.’’ The longer the ‘‘dead leg’’ is, the higher is degree temperature at all times. Because of the control system
of turbulence needed to keep it hot. delays inherent in its design, the purified water tem-
Although hot storage and distribution systems simi- perature will fluctuate by a few degrees every time a
lar to the one shown in Fig. 4 above are excellent major use point valve opens or closes. This is normally
from the standpoint of microbial control, their main not a problem, especially in the applications involving
limitation is the fact that water is delivered to all users filling the water into a process vessel. The heater in the
at the high (65–85 C) temperature. Very often, this is ambient loop (Fig. 7) is provided for periodical loop
not acceptable for some or all users. Other disadvan- sanitization; the steam supply is shut off at all other
tages of the hot systems are the higher potential for times.
corrosion of the construction materials, and personnel A third approach is to use the system similar to
safety concerns related to the high water temperature. the one shown in Fig. 4, but to keep the water in the
Various design concepts have been developed to
provide the users with purified water at required
(usually ambient) temperature.[12] Two of them are
shown in Figs. 6 and 7. In the first case (Fig. 6), water Purified water
Steam

is cooled directly at the point-of-use by a local cooler; from WFI still

in the second case (Fig. 7), a separate ambient tempera- Vent
Heater
ture loop is created, where one central cooler serves
Cond

the needs of multiple use points. During the periods Storage

tank
of no water usage, the cooler in the system in Fig. 7 80˚C
does almost nothing (just compensating for the heat
Steam

gains from the pump). However, once a point-of-use

Points of use
valve is opened and ambient water starts flowing out Heater
Circulation pump
of the loop, it is replaced by hot water from the storage
Cond

tank. At this point, the cooler needs to be able to

remove a lot of heat to keep the set loop temperature. Chilled
Water–Zeta

water
That usually requires a very large sanitary heat return
exchanger and a lot of cooling water. From the Cooler Points of use for
cooled water
practical prospective, it is often beneficial to install Chilled
two exchangers in series: the first one uses cooling Circulation pump water
supply
tower water to remove most of the heat, whereas the
second (‘‘trim cooler’’) uses more expensive chilled Fig. 7 Purified water storage and distribution system with
water to bring the loop to required temperature ambient loop.
 4046 Water for Pharmaceuticals

storage tank and in the circulation loop at ambient by creating an electrical arc between the weld electrode
temperature most of the time (this requires an extra and the tubing pieces, melting the ends of the tubes
heat exchanger with cooling water on the shell side). being joined. Unlike conventional welding, this tech-
To achieve microbial control, such system (or any nique creates a very smooth weld containing only the
system that includes ambient loop) needs to be period- materials from the joined pieces and no metal from
ically sanitized by heating up its contents to about the welding electrode. To avoid excessive oxidation
80 C and holding the temperature for at least 1 hr. of the weld area, it has to be purged with inert gas such
The sanitization frequency can vary from once a day as argon during the procedure. In addition to reducing
to once a week or less: the lower the frequency is, the the manual labor, the automatic welding machines
more attention to the studies proving the effectiveness help in maintaining a consistent quality of all welds,
of the microbial control shall be expected. as well as in documenting parameters (electrical
An alternative approach to microbial control in the current, electrode speed, purge flow, etc.) affecting such
ambient systems is to use ozone as a sanitizing agent. quality. Where automatic welding is not feasible (such
In such case, ozone gas (generated onsite from com- as, for example, in the fabrication of zero-static valves),
pressed air or oxygen) is continuously injected into manual welding is allowed; however, it is recom-
the purified water storage tank. An UV light fixture mended that it be conducted in the controlled shop
is installed at the outlet of the circulation pump to environment rather than at the construction site.
decompose ozone in the water directed to the users Welds in sanitary piping are often inspected by
during normal operation. Periodically, this UV light borescoping.
is shut off for 30 min to 1 hr, allowing ozonated water To improve the corrosion resistance of the stainless
to circulate through the distribution piping, thereby steel components, particularly in the weld-affected
achieving its sanitization. areas, it is a common practice to passivate the product
In larger systems, it often becomes impractical to contact surfaces of installed piping and equipment by
run one large-diameter piping loop to all use points; circulating a solution of strong oxidant such as nitric
in such cases, various multiloop arrangements may or citric acid, or a chelating agent. Passivation removes
be employed.[13] free iron from the metal surface and creates a thin pro-
The selection of a particular system design depends tective layer of chromium oxide. It is not uncommon
on many factors: number of use points requiring to repeat the passivation treatment periodically (every
ambient and hot water, flow rates and frequency of few years) to restore the passive layer.
usage, availability of capital, etc.[14] In the later years, some of the systems are designed
with the plastic materials of construction instead of the
stainless steel. The most common is polyvinylidene
Materials of Construction fluoride (PVDF); it is a partially fluorinated polymer
that exhibits many desirable properties (mechanical
Storage tank, piping, pumps, and other components of strength and chemical resistance) approaching those
the system in contact with purified water should be of the fully fluorinated Teflon, but is less expensive
made out of materials that are chemically resistant to and better suited for the installation in piping systems.
such water and which will not introduce metal ions Properly designed and supported PVDF piping can
or other contaminants. The most common material withstand hot water or even low-pressure steam sani-
used in the pharmaceutical industry is 316L stainless tization, as well as periodical exposure to ozone.
steel. The suffix ‘‘L’’ in the grade designation stands Although the PVDF tubing itself costs about as much
for ‘‘low carbon,’’ which is important for the compo- as the stainless steel and requires continuous support in
nents that need to be welded because carbon molecules most cases, it offers advantages such as light weight,
tend to diffuse to the surface during welding, making easier welding, less expensive valves and fittings, and
weld areas susceptible to corrosion. The piping systems no need for passivation. These benefits may result in
for purified water are usually constructed of the overall installed cost savings compared to the stainless
sanitary tubing. This product has tighter dimensional steel systems. Because of the concern about the leach-
tolerances than piping, and is available with the wide ing of organic impurities from plastics, researchers
range of surface finishes, up to the mirrorlike surfaces and suppliers of the sanitary PVDF tubing conducted
Water–Zeta

achieved by the combination of mechanical polish- extensive studies proving that the amount of TOC
ing and electropolishing. It is common to use the leached from such tubing into water is very low and
tubing polished to 25–30 min. roughness average (RA) does not degrade the water quality.[16,17] It is fair to
or better.[15] say that whereas none of the construction materials
The tubing and other components (valves, elbows, can be considered ideal for the purified water systems,
tees, etc.) are welded for the most part by the auto- plastics such as PVDF emerge as strong competitors to
matic orbital welding machines. Such machines work the stainless steel.[18]
Water for Pharmaceuticals 4047

Piping Design and Installation procedure. The piping is usually sloped a minimum
of 1/8 in/ft (or about 1 : 100) toward the nearest drain
While designing and installing the piping system for point to ensure complete drainage. Keep in mind that
purified water, care shall be taken to avoid any areas the thin-walled tubing used for the sanitary systems
where water would remain stagnant (‘‘dead legs’’). may sag between the support points, which would
Where branch connections are necessary, such as at create undrainable pools of liquid in the horizontal
the points of use, instrument ports, etc., the length of runs. Technically speaking, this sloping requirement
the ‘‘dead leg’’ is usually limited to a maximum is only important for the lines sanitized by steam (the
of three pipe diameters of the branch line, as measured condensate needs to drain freely to achieve even
from the main line wall. The importance of this ‘‘rule heating of all pipe surfaces), but is has become such
of thumb’’ in this particular application is in the fact a common practice that the author is not aware of
that while the piping system is being sanitized (either any purified water systems in the pharmaceutical
periodically or continuously), all parts of it, including industry installed without slope. Most of the piping
the most remote areas, have to reach the required and other system elements are welded, and those com-
temperature, or, in case of chemical sanitization, be ponents that may require disassembly (pumps, heat
accessible for that chemical. In cases where long ‘‘dead exchangers, instruments, etc.) are connected by sani-
legs’’ cannot be avoided (such as in some existing sys- tary joints such as Tri-Clamp, Swagelock TS, etc.
tems or in smaller new systems), microbial control can The common feature of all such joints is that gasket fits
be achieved by flushing each of such legs with hot flush to the tube wall, making the joint crevice-free.
water or chemical solution as part of the sanitization Diaphragm valves have become a de facto standard
for purified water as well as for most other sanitary
systems because of their cleanability, absence of crevices,

Water–Zeta

Fig. 8 Weir-style diaphragm valves: shutoff and point-of- Fig. 9 Radial-style diaphragm valves: shutoff and flow-
use configurations. (Courtesy of ITT Industries.) through configurations. (Courtesy of NovAseptic.)
 4048 Water for Pharmaceuticals

and complete isolation of the wetted parts from the 3. Weitnauer, A.; Comb, L. Reverse osmosis for WFI and
actuator mechanism. The two most widely used styles PW. Presented at Pharmaceutical Waters’95, Atlantic City,
NJ, May, 1995.
are weir and radial diaphragm valves, respectively. In 4. Schaule, G.; Flemming, H. Pathogenic microorganisms in
addition to simple (one inlet, one outlet) valve con- water system biofilms. Ultrapure Water 1997, 14 (4), 21–25.
figuration, several companies offer a wide variety of 5. Gillis, R.; Gillis, J. A comparative study of bacterial attach-
ment to high-purity water system surfaces. Ultrapure Water
point-of-use and other specialty valves designed for 1996, 13 (6), 27–32.
the pharmaceutical and related industries in general, 6. Riedewald, F. Biofilms in pharmaceutical waters. Pharm.
and for the purified water systems in particular Eng. 1997, 17 (6), 8–19.
7. Collentro, W. Microbial control in purified water systems—
(Figs. 8 and 9). Case histories. Ultrapure Water 1995, 12 (3), 30–36.
8. Byrne, W. Biological control in high-purity water. Pre-
sented at Ultrapure Water Expo, Philadelphia, PA, 2001.
9. ISPE Baseline Pharmaceutical Engineering Guides: Vol. 4.
CONCLUSIONS Water and Steam Systems; ISPE, 2001.
10. Gray, G.C. Recirculation velocities in water for injection
distribution systems. Pharm. Eng. 1997, 17 (6), 28–33.
There are many types of purified water systems used in 11. Klauer, J. An examination of pipe self-cleaning in
pharmaceutical facilities. Although most of them share high-purity water systems. Ultrapure Water 2001, 18 (3),
common features, each system is custom-designed for 56–60.
12. Shnayder, L. Pharmaceutical purified water storage and dis-
a specific application. Developing a proper design tribution systems—an engineering prospective. Pharm. Eng.
requires a good understanding of system operation 2001, 21 (6), 66–72.
and careful attention to details. Simply following com- 13. Dreyer, C.; Du, S.; Sommer, K.; Williams, R. Criteria, tools,
and practices for high-purity water distribution systems.
mon ‘‘rules of thumb’’ does not necessarily guarantee a Ultrapure Water 2000, 17 (5), 17–28.
reliable system—no matter how much money is spent. 14. McWilliam, A. The design of high purity water distribution
On the other hand, with a good understanding, it is systems. Pharm. Eng. 1995, 15 (5), 54–70.
15. Peterman, L. Design guidelines for WFI and other high-
often possible to design, install, and validate a func- purity water systems. Presented at ISPE Conference on
tional and reliable purified water system with less Pharmaceutical Water Systems, Amsterdam, The Nether-
capital investment and lower operating costs. lands, 1991.
16. Motomura, Y.; Yabe, K. Piping materials and distribution
systems for advanced ultrapure water. Proceedings of Tenth
Annual Semiconductor Pure Water Conference, Santa
Clara, CA, 1991, 1–22.
REFERENCES 17. Govaert, R.; Lueghamer, A. Thermoplastic piping systems
for pharmaceutical water applications. Ultrapure Water
1. United States Pharmacopeia 26 National Formulary 21; US 2001, 18 (10), 32–39.
Pharmacopeia Convention Inc.: Rockville, MD, 2003. 18. Sixsmith, T.; Jackson, J. How piping materials for the phar-
2. Meltzer, T. Pharmaceutical Water Systems; Tall Oaks maceutical industry compare to each other. Ultrapure
Publishing: Littleton, CO, 1997. Water 1999, 16 (4), 53–59.
Water–Zeta
Water Sorption of Drugs and Dosage Forms
Mark J. Kontny
Pharmacia, Skokie, Illinois, U.S.A.

James J. Conners
Sepracor, Marlborough, Massachusetts, U.S.A.

INTRODUCTION environmental relative humidity is significantly differ-

ent from the relative humidity at which the excipient
The physical, chemical, and mechanical properties of (gelatin, for example) was previously equilibrated, the
pharmaceutical drugs and dosage forms are critically initial moisture uptake/loss rate will be significant,
dependent on the presence of moisture. Pharmaceutical but it will approach zero over time. On the other hand,
scientists can cite numerous examples of desirable and a water-soluble, non-hydrating crystalline substance
undesirable properties that result from varied levels of such as sodium chloride will have a very low moisture
moisture associated with a particular solid or formu- uptake/loss rate that will decrease to zero if the
lation consisting of mixtures of solids. Flow, compac- environmental relative humidity is kept below its criti-
tion, caking, disintegration, dissolution, hardness, and cal relative humidity. However, the uptake rate will be
chemical stability are just some of the properties relatively large and continuous, until all the solid has
influenced by moisture. Because water is present in bulk dissolved, if the relative humidity is above the critical
liquid form or as a vapor at some relative humidity in value. Obviously, very different mechanisms of water
virtually all stages of solid manufacture (active ingredi- sorption/desorption occur for the different samples.
ent and excipients), storage, processing into formula- In this light, describing sodium chloride and/or starch
tions, and final product packaging, a fundamental as ‘‘hygroscopic’’ offers very little toward understand-
understanding of the role of water in affecting solid ing the water–solid interactions that might affect their
properties (and vice versa) is necessary. physico-chemical properties. These examples illustrate
Although the properties of individual solids and the the need to understand the underlying mechanism(s)
performance of solid dosage forms are dependent on of uptake for a particular solid. In this regard, there-
moisture, characterization of the underlying water– fore, addressing the following questions provides a
solid interaction is often nebulous. For example, many basis for studying the various mechanisms of water–
solids are described as ‘‘hygroscopic’’ without further solid interaction:
reference to whether and how this relates to the rate
and amount of moisture uptake as a function of rela-
tive humidity and temperature.[1] To illustrate this 1. How much water is present and what is the
ambiguity, consider that water-soluble, non-hydrating corresponding water activity (approximated by
crystalline substances such as sodium chloride sorb relative pressure or percent relative humidity/
very low levels of moisture (e.g., less than 0.1%) below 100)?
their critical relative humidities, where the solid actu- 2. What are the kinetics of moisture uptake or loss,
ally dissolves in the sorbed moisture. On the other and is the rate constant or changing over time?
hand, some typical excipient materials used in solid 3. Where is the water located (i.e., adsorbed to the
dosage forms, such as starches, celluloses, and gelatin external surface of crystals, absorbed into
capsules, sorb significant quantities of moisture (e.g., crystals as specific or non-specific water of
25–50%), and even though they do not dissolve, they hydration, absorbed into amorphous regions,
do undergo significant morphological change at high condensed into pores, etc.)?
relative humidities (i.e., swelling). Moisture uptake rate 4. What is the state of the moisture associated with
for a material depends on both the relative humidity of the solid (i.e., bulk water, water of hydration,
Water–Zeta

the environment and the time-dependent moisture physisorbed water, etc.)?

content of the solid. For situations in which the 5. What form of the solid is present (i.e., particle
size and morphology, polymorphic species,
degree of crystallinity, state of hydration) and
This work was taken in part from the article entitled ‘‘Sorption of
is this form thermodynamically stable over the
water by solids,’’ in Physical Characterization of Pharmaceutical temperature and relative humidity range that
Solids, Brittain, H., Ed., Marcel Dekker, Inc.: New York, 1995. the solid is expected to encounter?
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-100200016
Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved. 4049
 4050 Water Sorption of Drugs and Dosage Forms

It is the objective of this article to highlight the where P is the water vapor pressure in the system and
various mechanisms whereby water can interact with P0 is the vapor pressure above pure water at the tem-
solid substances, present methodologies that can be perature of interest. Relative humidity (RH) is defined
used to obtain the necessary data, and then discuss as the relative pressure expressed on a percentage basis:
moisture uptake for non-hydrating and hydrating
crystalline solids below and above their critical relative P
RH ¼ 100 ð4Þ
humdities for amorphous solids and for pharmaceuti- P0
cally processed substances. Finally transfer of moisture
from one substance to another will be discussed. The sorption branch of the isotherm is obtained exper-
imentally by measuring the equilibrium amount of
water sorbed to a solid at known relative pressure,
beginning with a known mass of absolutely dry solid
THE WATER SORPTION ISOTHERM
and then progressively increasing the relative pressure
in the system. Drying the solid sample under heat,
The most fundamental manner of demonstrating the
possibly using vacuum to facilitate the removal of des-
relationship between sorbed water vapor and a solid
orbed water vapor, is usually necessary to eliminate
is the water sorption–desorption isotherm. The water
residual moisture. One must be aware, however, of
sorption–desorption isotherm describes the relation-
the effects of such conditions on the chemical and
ship between the equilibrium amount of water vapor
physical stability of the solid. The desorption portion
sorbed to a solid (usually expressed as amount per unit
of the isotherm is obtained by progressively decreasing
mass or per unit surface area of solid) and the thermo-
the relative pressure in the system from a relative press-
dynamic quantity, water activity (aw), at constant tem-
ure of approximately unity, again monitoring the equi-
perature and pressure. At equilibrium the chemical
librium amount of moisture sorbed at each relative
potential of water sorbed to the solid must equal the
pressure. Remember that the moisture sorption iso-
chemical potential of water in the vapor phase.
therm is an equilibrium measurement of the interaction
Water activity in the vapor phase is related to chemical
of water with a solid. In theory, information regarding
potential by
the kinetics of moisture uptake is not explicitly derived
from this experiment. This distinction is an important
m ¼ m0 þ RT ln aw ð1Þ
one that will be explored in more depth later.
Generation of water sorption–desorption isotherms
where m is the chemical potential of water in the system
for a particular solid can lend considerable insight into
at equilibrium, m0 is the standard chemical potential of
the nature of the water–solid interaction, as well as the
water at a specific reference temperature and pressure,
surface characteristics of the solid. This information is
R is the gas constant, and T is the absolute tempera-
readily obtained from the amount of moisture sorbed
ture. Lewis et al.[2] defined the relative activity of any
at lower relative humidities in comparison with the
pure substance or component (such as water) as a ratio
specific surface area of the sample, from the general
of fugacities:
shape of the isotherm, from whether or not water
fw uptake is a completely reversible process (i.e., whether
aw ¼ ð2Þ hysteresis is observed between sorption and desorp-
fw0
tion), and from the shape of the hysteresis loop if it
where fw is the fugacity of water in the system at is present. With knowledge of the aforementioned,
equilibrium and f 0w is the fugacity of pure water at a one can usually obtain an indication of the mechanism
standard temperature and pressure. For all practical of moisture sorption for the material of interest. For
purposes, the fugacity (or ‘‘escaping tendency’’) of example, a material that exhibits sorption at lower
water vapor can be approximated by the water vapor relative humidities in much greater amounts than one
pressure in the system. This assumption is valid as long might expect based on the specific surface area of the
as the water vapor behaves as an ideal gas. For the sample, and that exhibits hysteresis over the complete
water vapor pressure range of relevance for pharma- range of relative humidities, is most likely absorbing
ceutical systems at temperatures less than 50
C, this water into its internal structure. On the other hand, a
Water–Zeta

approximation is excellent (<0.2% relative error).[3] material exhibiting a closed hysteresis loop over the
Thus, the relative pressure of water vapor, P/P0, is higher relative humidity range while sorbing moisture
usually employed as an estimate of the relative water over the lower relative humidity range similar to what
activity in the system: might be expected based on its specific surface area,
is probably quite porous in nature and is most likely
P sorbing water via capillary condensation over the
aw ¼ ð3Þ
P0 higher relative humidity range.
Water Sorption of Drugs and Dosage Forms 4051

MODELS DESCRIBING VAPOR ADSORPTION relative pressures than is experimentally observed. This
is consistent with an assumption built into the BET
Brunauer, Emmett, and Teller Equation derivation that an infinite number of layers are
adsorbed at a relative pressure of unity. Application
The model most commonly referred to in the literature of the BET equation to non-polar gas adsorption
describing vapor adsorption onto solid surfaces was results is carried out quite frequently to obtain esti-
put forth in 1938 by Brunauer, Emmett, and Teller.[4] mates of the specific surface area of solid samples.
The so-called BET model was originally derived using By assuming a cross-sectional area for the adsorbate
kinetic arguments in a manner very similar to those molecule, one can use Wm to calculate specific surface
used by Langmuir.[5] The BET model has since also area by the following relationship:
been derived using statistical mechanics.[6–8] The BET
model assumes that vapor molecules, behaving as an Wm XNav
S ¼ ð9Þ
ideal gas, exist in a state of equilibrium with a solid MS
that consists of identical, homogeneous adsorption
sites. The first vapor molecule adsorbed to an adsorp- where S is the specific surface area in m2/g; Wm is the
tion site on the solid is proposed to be bound, whereas mass of adsorbate adsorbed at monolayer coverage;
molecules adsorbing beyond the first layer are assumed X is the cross-sectional area of an adsorbed adsorbate
to have the properties of the bulk liquid. Furthermore, molecule (assumed to be 19.5 Å2 for krypton, 16.2 Å2
adsorption is proposed to occur such that the adsorbed for nitrogen, 12.5 Å2 for water);[9,10] Nav is Avogadro’s
molecules do not interact laterally. The linear form of number; M is the molecular weight of adsorbate; and S
the BET equation is is the mass of the sample. Obviously, calculating sur-
face areas from moisture uptake data that does not
1 ðCb  1Þ  P
1 lead to monolayer coverage at Wm [either incomplete
P0
 P0
 ¼ þ ð5Þ coverage (see the section on ‘‘Water Sorption onto
W P1
Wm C b W m Cb
Non-hydrates’’ below) or absorption into the solid
(see the section on ‘‘The Meaning of Specific Surface
where W is the mass of vapor adsorbed per gram of Areas Calculated from Water Absorption Studies’’)]
solid at a particular relative pressure, P/P0; Wm is the will result in incorrect values that have no physical
theoretical quantity of vapor adsorbed when each meaning. Therefore, comparison of the surface area
adsorption site has one vapor molecule adsorbed to measured by non-polar gas adsorption to that calcu-
it; and lated from a moisture sorption isotherm can lend
  insight into the fundamental interactions between the
H1  HL
Cb ¼ k exp ð6Þ water and the solid. This will be explored in some
RT
depth later.

where H1 is the heat of adsorption of the first vapor

molecule adsorbed to a site, HL is the heat of con-
Guggenheim and deBoer Equation
densation of the bulk adsorbate, R is the universal
gas constant, T is the absolute temperature, and k is
Many attempts to modify the BET adsorption theory
a constant, usually assumed to be close to unity. The two
have been made since its original derivation. Its sim-
BET constants, Wm and Cb, can easily be obtained from
plicity and ability to fit adsorption data extremely well
the linear plotting form of the BET equation given in
at lower relative pressures, however, have made it the
Eq. (5). Plotting the quantity 1/[W(P0/P  1)] vs. P/P0
model of choice for estimating surface areas from
gives a slope equal to (Cb  1)/WmCb and an intercept
non-polar gas adsorption. Most modifications of the
equal to 1/WmCb. Algebraic manipulation gives
BET model, developed to analyze data over the entire
1 range of relative pressures, usually add at least one fit-
Wm ¼ ð7Þ ting parameter to the equation. This makes computer
slope þ intercept
fitting a necessity, because only two measurable para-
and meters, W and P/P0, are available. From a modeling
Water–Zeta

perspective, additional fitting parameters of unknown

slope or undefined physical meaning that arise from such
Cb ¼ 1 þ ð8Þ approaches are often a deterrent to the use of multi-
intercept
parameter models because of the consequent difficulty
In general, the BET equation fits adsorption data in interpreting results. In this regard, therefore, only a
quite well over the relative pressure range 0.05–0.35, single modification of the BET model, which has been
but predicts considerably more adsorption at higher shown to extend the relative pressure range over which
 4052 Water Sorption of Drugs and Dosage Forms

vapor adsorption data can be fit, will be considered of P/P0 using its extension, Eq. (10). Because this
here. This extension of the BET model, independently was first reported by Anderson[18] to be the case for
derived by Guggenheim,[11] and deBoer,[12] accounts water absorption, Eq. (10), when applied to water
for the adsorption of an intermediate state of vapor vapor sorption, is often called the GAB equation for
between the tightly bound first molecule adsorbing Guggenheim, Anderson, and deBoer.[19] Because the
to an adsorption site and the condensed molecules theoretical process for the derivation of the original
adsorbed at very high relative pressures. Molecules equation does not translate directly to the absorption
adsorbed in the intermediate range can be considered process, which involves dissolution of water in the
to interact with the solid, but the interaction is amorphous solid, the significance of fit to the GAB
assumed to be considerably less than that of the first equation is somewhat limited. It is, however, a very
molecule sorbed at an adsorption site. Obviously, this useful equation because it does allow one to describe
addition of a ‘‘third state’’ of interaction is an approxi- the entire isotherm and to draw out some useful param-
mation. In all likelihood, there is a continuum of inter- eters (to be discussed in what follows).
action states. However, from a computational point of Since water vapor dissolves in the solid during
view, the existence of three or more states is indis- absorption, several models based on solution theory,
tinguishable. The equation for the three-state interac- proposing that the sorbate is taken up into the solid
tion model is given as as a solid solution, have been derived and used to
describe water sorption on polymers (e.g., Flory-
Wm CG K PP0 Huggins,[20] Hallwood-Horrobin[21]). More recently,
W ¼     ð10Þ Vrentas et al.[22,23] developed a solution-based model
1  K PP0 1  K PP0 þ CG K PP0
that accounts for the plasticizing effect of water on a
where P, P0, HL, W, and Wm are identical to the param- polymer that has been shown to describe the entire
eters used in the BET equation, and moisture uptake isotherm for the polymer.[24] While
these sorption theories and the many modifications
  of the BET adsorption model are based on meaning-
HL  Hm ful physicochemical principles, further work is still
K ¼ B exp ð11Þ
RT required to elucidate the molecular mechanisms under-
lying moisture uptake into polymeric systems. From
where B is a constant and Hm is the heat of adsorption this perspective, other models based on entirely dif-
of vapor adsorbed in the intermediate layer. The con- ferent theoretical concepts will not be considered
stant CG is defined as further in this article. For further reference, the reader
is directed to several excellent literature reviews of the
CG ¼ D expðH1  Hm RT Þ ð12Þ many sorption theories that have been proposed.[25,26]

where D is a constant, H1 is the heat of adsorption of

the first molecule adsorbed at a site, and Hm is the heat Capillary Condensation
of adsorption of the intermediately bound molecule.
Vapor sorption onto porous solids differs from vapor
uptake onto the surfaces of flat materials in that a
Water Vapor Absorption by vapor (in this case, water) will condense to a liquid in
Amorphous Solids a pore structure at a vapor pressure, Pr, below the
vapor pressure, P0, where condensation occurs on flat
The process of water vapor interaction with amorph- surfaces. This is generally attributed to the increased
ous solids has been likened to the production of a solid attractive forces between adsorbate molecules that
solution in which the water is dissolved in the solid occur as surfaces become highly curved, such as in a
matrix. As more water is absorbed, the fundamental pore or capillary. This phenomenon is referred to as
properties of this solid matrix (e.g., viscosity) can capillary condensation and is described by the Kelvin
undergo significant change that can then result in visu- equation:[27]
ally apparent changes in physical properties (e.g., col-  
Water–Zeta

lapse, recrystallization).[13–17] This will be discussed in Pr

ln 0 ¼ 2gVm rRT ð13Þ
further detail later. P
Although water vapor is absorbed into amorphous
solids and not simply adsorbed on the surface, it still where g is the surface tension of the adsorbed film
has been found that such sorption isotherms can be (assumed equal to that of the bulk liquid), Vm is the
fit to the BET equation up to a P/P0 of about 0.40 molar volume of the liquid, r is the pore radius, R is
as with vapor adsorption, and over the entire range the gas constant, and T is the temperature. The Kelvin
Water Sorption of Drugs and Dosage Forms 4053

equation has been shown to be applicable to pore radii temperature. Because relative humidity is dependent
as low as 5 nm for water adsorption onto mica.[28,29] As on the number of dissolved species, it is essential that
mentioned in the section on ‘‘The Water Sorption Iso- saturation be attained prior to beginning experimen-
therm,’’ capillary condensation will result in a closed tation. In this regard, preparing the salt solutions
hysteresis loop in the adsorption/desorption isotherm several days before beginning a sorption study is
of a porous material. Calculating Pr/P0 by assuming recommended. Sulfuric acid solutions of varying
a surface tension of water of 72.8 ergs/cm2 and a den- concentration[35] are also used to establish relative
sity of 0.998 g/cm3 at 293 K (this is an assumption for humidity. Addition or removal of water from the solu-
the purposes of this calculation as it has been shown tion by desorption or sorption of water to the solid,
that the density of water is lower in a pore than the however, will alter the concentration of sulfuric acid
bulk value)[30] shows that condensation is predicted (and water) in solution, and thus change the relative
at relative pressures of 0.998, 0.989, 0.898, and 0.340 humidity of the headspace. This technique for control-
for pore radii of 1000, 100, 10, and 1 nm, respectively. ling relative humidity in the headspace is practically
In this regard, it is clear that capillary condensation more useful when small amounts of water are sorbed/
need only be considered for very small pore dimen- desorbed from the solid.
sions. In practical terms, one should be concerned Temperature modification of an aqueous solution
about this mechanism of water uptake for microporous can also be used to maintain constant relative humidity
pharmaceutical powders that exhibit a relatively large in the headspace.[19] This technique maintains the solid
specific surface area (i.e., >100 m2/g), as determined at one temperature and an aqueous solution connected
from non-polar gas adsorption studies. to the system at another temperature. Due to strong
vapor pressure dependence on temperature, very tight
temperature control of the aqueous solution and the
METHODOLOGY solid are required to maintain constant relative
humidity in the vicinity of the solid by this technique.
Control of Relative Humidity Control of the vapor pressure in the headspace over
a solid can also be used to maintain a relative humidity
Maintenance of constant relative humidity environ- over a solid. As shown in Eq. (4), the relative humidity
ments is essential for studying water–solid interactions. is directly correlated to the partial pressure of water in
There are primarily four techniques that are frequently the vapor phase. To utilize this technique for relative
employed to maintain constant relative humidity: humidity control, the headspace above the sample
must be completely evacuated prior to analysis. Pure
1. Colligative solutions water vapor can then be carefully admitted to the
2. Temperature modification of an aqueous vapor phase. Because only water vapor is present, the
solution pressure measured over the system is directly related
3. Control of total pressure over the solid to the relative humidity over the sample.[36]
4. Mixing wet and dry air streams Mixing dry and water vapor saturated air in defined
proportions also can be used to generate constant rela-
Saturated salt solutions and sulfuric acid solutions tive humidity. Control of flow rates and the water
establish relative humidity by reducing the vapor press- vapor content of the dry and saturated air are essential
ure above an aqueous solution (a colligative effect). to ensure accurate, reproducible relative humidity
Saturated salt solutions at controlled temperature production.[37,38]
maintain a constant relative humidity as long as excess
salt and bulk solution are present. As water is added or
removed from the solution, moisture from the head- Measurement of Relative Humidity
space will either condense or evaporate (as appropri-
ate), with subsequent dissolution or precipitation of Measurement of relative humidity depends on the sys-
salt to maintain the equilibrium vapor pressure. tem used. Systems employing vacuum are usually evac-
Because the degree of vapor pressure depression is uated prior to introduction of water vapor.[36,39] For
dependent on the number of species in solution cases in which a gas-forming reaction is not occurring,
Water–Zeta

and, further, since the solubility of most salts is some- measurement of total pressure in the system can be
what dependent on temperature, the relative humidity used as a measure of water vapor pressure. Systems
generated is also temperature dependent. Hence, use in which air is not evacuated require specific measure-
of the same salt at different temperatures can result ment of water vapor pressure. (For the latter type of
in different relative humidities. Refs.[31–35] can be system, caution should be taken to assure that the rela-
consulted for specific saturated salt solutions that tive humidity source is in close proximity to the solid,
result in defined relative humidities as a function of since the diffusion of water vapor through air to the
 4054 Water Sorption of Drugs and Dosage Forms

solid is required to maintain a constant relative carried out either gravimetrically or volumetrically.
humidity in the immediate vicinity of the solid.) A wide Gravimetric methods require:
variety of pressure measuring instrumentation is com-
mercially available with varying accuracy, precision, 1. A dry sample weight,
and cost. 2. Constant temperature of the sample,
3. Maintaining predetermined constant relative
humidities in the headspace, and
Measurement of the Critical Relative 4. Attaining and measuring an equilibrium weight
Humidity, RH0 of sorbed water vapor. Gravimetric measure-
ment of moisture uptake can occur continuously
The relative humidity at which a solid begins to deli- or discontinuously. Continuous measurement
quesce, RH0, can be determined in three ways: 1) usually involves placing a sample on a balance
directly, by measuring the relative humidity above a in a temperature- and relative humidity-
saturated solution of the substance; 2) by determining controlled environment. Microbalances in
the relative humidity at which significant moisture closed systems have been used successfully for
uptake and simultaneous dissolution occurs; or this purpose,[37–39,44] and commercial systems
3) indirectly, by measuring the steady state moisture are now available that can accurately and
uptake rate at relative humidities above RH0 and then precisely control relative humidity and simul-
extrapolating to the relative humidity at which the taneously monitor sample weight.
moisture uptake rate is zero.[1,40,41]
Although other techniques can be used to measure Volumetric methods require:
the relative humidity above a saturated solution, one
relatively simple procedure is to evacuate the head- 1. A dry sample weight,
space (to remove air by vapor phase expansions) and 2. Constant temperature of the sample,
then, with the vacuum pumps isolated and the satu- 3. Water vapor pressure measurement in a dosing
rated solution maintained at a constant temperature, volume and, later, in the headspace above the
to measure water vapor pressure. Water vapor pressure equilibrated sample, and
can then be converted to relative humidity by dividing 4. Measuring dead volumes of the individual cham-
by P0, the vapor pressure above pure water at the bers, including the sample chamber. In essence,
temperature of interest.[42] volumetric methods equilibrate a known head-
space dosing volume at a given (measured) water
Measurement of Moisture Uptake vapor pressure, followed by exposure of the
(Kinetics of Deliquescence) pre-equilibrated sample to this water, with sub-
sequent measurement of the water vapor pressure
The rate of moisture uptake above RH0 requires main- after equilibration. The mass of water sorbed, Dn
tenance and measurement of a range of relative humid- (in moles), at the final pressure in the system, Pf,
ities, and the capability of measuring the moisture is obtained from the difference, DP, between Pcalc
f ,

content of the solid over time. Use of a vacuum system the calculated water vapor pressure at equilib-
can minimize vapor diffusion through the headspace, rium, and Pmeas
f , the final measured water vapor
thus maintaining constant relative humidity in the pressure:
vicinity of the sample. Also, because the most reliable DPV
estimate of the steady-state moisture uptake rate is Dn ¼ ð14Þ
RT
when the integrity of the solid is intact and the film
of sorbed moisture is thin (and saturation most likely), where V is the final volume, R is the gas constant,
it is advisable to determine the moisture uptake rate and T is the absolute temperature.[39]
at early time periods. In this regard, it is also helpful
to be able to view the solid during the experiment to
verify that integrity is maintained and excess solid
remains.[41,43] WATER SORPTION BY
Water–Zeta

CRYSTALLINE SOLIDS

Measurement of Equilibrium General Model

Moisture Sorption
Fig. 1 schematically describes the important steps in
Generation of water sorption/desorption isotherms in the uptake of water vapor by crystalline water-soluble
a controlled relative humidity environment can be solids. At low relative humidities, water is adsorbed to
Water Sorption of Drugs and Dosage Forms 4055

A B C (Teflon),[29] but it sorbs to a greater extent to more

polar materials such as alkali halides[45–48] and organic
Adsorption Dissolution Deliquescence
salts like sodium salicylate.[48] Because water is only
sorbed to the external surface of these substances, rela-
Solution tively small amounts (i.e., typically less than 1 mg/g) of
water are sorbed compared with hydrates and amorph-
ous materials that absorb water into their internal
Solid Solid Solid structures.
Unfortunately, the literature is relatively sparse with
examples showing the water uptake profile onto crys-
talline, non-hydrating substances below RH0. This is
Fig. 1 Water vapor adsorption and deliquescence of a most likely due to the difficulty in accurately measur-
water-soluble solid at (A) atmospheric relative humidity, ing the small amounts of water that are sorbed. Alkali
RHi < RH0; (B) RHi ¼ RH0; (C) RHi > RH0. halides are an exception, however, likely due to their
well-characterized particle morphologies.[45–48] Fig. 2
the surface of a non-hydrate-forming solid. As the rela- shows a water uptake isotherm onto recrystallized
tive humidity is increased, some tendency for multi- sodium chloride.[48] Note that the amount of water
layer sorption is expected. At some relative humidity sorbed as a function of relative humidity is normalized
(characteristic for a given substance), the solid will to the specific surface area of the sample. Because
begin to dissolve in the sorbed film of water. A satu- water is sorbed only to the external surface of this
rated solution of solute will most likely exist, and this material, this allows comparison of water uptake data
will cause the vapor pressure over the sorbed film of from different lots of material, whereas plotting this
water to be depressed relative to pure water and to
be constant and equal to that above a saturated
solution of the substance. This vapor pressure may 60
be expressed as the critical relative humidity, RH0. If
the relative humidity in the atmosphere is greater than
that over the saturated solution (RH0), water will
spontaneously condense on the aqueous film. This will 50
dilute the film allowing more solid to dissolve, which,
in turn, will maintain the pressure gradient. The pro-
cess of water vapor uptake will continue until all the
W ×10×5 (G water / square meter)

solid has dissolved and further solution dilution has

40
occurred. Only when the relative humidity above the
solution is elevated to that of the atmosphere will this
process terminate. This phenomenon is called deli-
quescence. Although hydrates undergo solid state tran-
sitions in transforming from the anhydrate to hydrate, 30
as well as from one hydrate species to another, beha-
vior similar to that previously described for non-
hydrates is also noted at and above RH0 for hydrates.
In pharmaceutical systems, water-soluble species are 20
frequently encountered in dosage forms. Thus, it is
important to understand the conditions responsible
for deliquescence and the molecular events occurring
at relative humidities below the deliquescence point.
10

Water Sorption onto Non-Hydrates

Water–Zeta

below RH0
0
The sorption of water vapor onto non-hydrating crys-
0.0 0.2 0.4 0.6 0.8
talline solids below RH0 will depend on the polarity of
Relative pressure
the surface(s) and will be proportional to surface area.
For example, water exhibits little tendency to sorb to Fig. 2 Water vapor sorption for recrystallized (&) and
non-polar solids like carbon or polytetrafluoroethylene ground (Z) sodium chloride at 20
C. (From Ref.[48].)
 4056 Water Sorption of Drugs and Dosage Forms

data on a ‘‘per gram’’ basis would have little or no state. Below this characteristic relative humidity, these
meaning. For the sodium chloride sample in Fig. 2 materials behave similarly to non-hydrates. Once the
(specific surface area ¼ 0.0875 m2/g from krypton characteristic relative humidity is attained, addition
adsorption studies), only 5  104 g water/m2 of of more water to the system will not result in a further
sodium chloride is sorbed, even up to 70% relative increase in relative humidity. Rather, this water will be
humidity. Also, note the apparent step-like nature of sorbed so that the anhydrate crystal will be converted
the isotherm. From BET analysis of the sorption to the hydrate. The strength of the water–solid interac-
data at the lower relative humidities, a Wm value of tion depends on the level of hydrogen bonding possible
7.6  105 g/m2 is obtained. This value is only about within the lattice.[29,49] In some hydrates (e.g., caffeine
0.32 that of the predicted value for monolayer coverage and theophylline) where hydrogen bonding is relatively
assuming an area per water molecule of 12.5 Å2. This weak, water molecules can aid in hydrate stabilization
suggests that it is quite meaningless to refer to the primarily due to their space-filling role.[29,49,50]
number of layers of sorbed water as multiples of Wm, Since water molecules occupy regular positions
except as a point of reference. Interestingly, the second within the lattice of a hydrate with a specific stoichi-
step plateau in Fig. 2 occurs at about three times the ometry (e.g., 1 : 1 monohydrate, 2 : 1 dihydrate, 5 : 1
moisture content corresponding to Wm, suggesting that pentahydrate) to the solid, relatively large quantities
for sodium chloride, the monolayer is actually com- of water are sorbed. Fig. 3 shows a moisture uptake
pleted during the second step of the isotherm. Isosteric isotherm for ipratropium bromide.[51] This substance
heat of sorption results for sodium chloride from undergoes an apparent hydration of the crystal
Barraclough and Hall[45] suggest that the heat of sorp- between 63% and 75% relative humidity. Above 75%
tion of water up to Wm is invariant, whereas the heat of relative humidity, approximately 4.6% water is sorbed
sorption decreases and becomes constant at about two (theoretical monohydrate is 4.4%). Interestingly, as
times Wm. Considering the experimental error involved anhydrous ipratropium bromide is equilibrated for
in obtaining Wm and the isosteric heats of sorption, this extended time periods (e.g., 2 and 5 months respectively,
suggests that water is sorbed with a homogeneous as shown in Fig. 3), hydration of the crystal appears to
binding energy up to Wm and then interacts to a lesser
extent until the monolayer is complete.
As shown in Fig. 2[48] and also in the work of 0.06
Barraclough and Hall,[45] moisture uptake onto sodium
chloride as a function of relative humidity is reversible
as long as RH0 is not attained. This is evidence
that actual dissolution of water-soluble crystalline sub- 0.05
stances does not occur below RH0. This is consistent
with the thermodynamic rationale that dissolution below
RH0 would require a supersaturated solution (i.e., an 0.04
W (G water / G solid)

increased number of species in solution would be neces-

sary to induce dissolution at a relative humidity below
that of the saturated solution, RH0). In this regard,
0.03
one should only need to consider the solid state proper-
ties of a purely crystalline material below RH0. As will
be described, other considerations may be warranted
for a substance that exists in multiple polymorphic forms 0.02
or contains amorphous material.

0.01
Water Sorption onto Hydrates below RH0

Many drugs and excipients (cephalexin monohydrate,

quinidine sulfate dihydrate, ampicillin trihydrate, 0.00
Water–Zeta

codeine sulfate trihydrate, morphine sulfate dihydrate, 0.0 0.2 0.4 0.6 0.8 1.0
dicalcium phosphate trihydrate, raffinose pentahy- Relative pressure
drate, lactose monohydrate) utilize water as an integral Fig. 3 Water vapor sorption and desorption isotherms for
part of their crystal structure. Solids that form specific ipratropium bromide at 20
C. (&) 2-month sorption results;
crystal hydrates tend to sorb relatively small amounts (`) 5-month sorption results; (Z) 2- and 5-month desorption
of water to their external surface below a characteristic results. (Note: All 2-month sorption results, except at 53%
relative humidity, when initially dried to an anhydrous and 63% relative humidity, were verified at 5 months.)
Water Sorption of Drugs and Dosage Forms 4057

occur at 53% and 63% relative humidity. This example the expected RH0 for a solute of significant aqueous
clearly shows that a time period of many months may solubility. Hence, RH0 should be measured for individ-
be required to attain a reliable estimate of the equilib- ual solids. Examples of RH0 values for single compo-
rium uptake at select relative humidities. Characteristic nent systems are shown in Table 1.
of many hydrates, ipratropium bromide exhibits signi- Values of RH0 for mixtures, on the other hand, can
ficant hysteresis between the sorption and desorption be calculated from the RH0 values of single compo-
isotherms. This is attributed to the degree of binding nents using an equation developed by Ross:[54]
and the physical fit of water in the hydrated lattice.
Non-specific hydration, or hydration of the lattice    
ðRH0 Þmix ðRH0 Þ1 ðRH0 Þ2
without a first-order phase transition, also must be ¼ 
100 100 100
considered. Cox, Woodard, and McCrone[52] reported  
the moisture uptake profile of cromolyn sodium, and ðRH0 Þ3
  ð15Þ
the related effects on the physical properties of this 100
substance. Although up to nine molecules of water
per molecule of cromolyn sodium are sorbed into the where (RH0)mix is the relative humidity above a satu-
crystalline lattice at 90% relative humidity, the sorp- rated solution of the mixture and (RH0)i represents
tion profile does not show any sharp plateaus corre- the relative humidities of the individual saturated salt
sponding to fixed hydrates. Rather, the uptake profile solutions. The Ross equation was derived assuming
exhibits a gradual increase in moisture content as dilute solutions and negligible interaction between the
relative humidity increases, which results in marked components in solution. The results presented in
changes in X-ray diffraction patterns, density, and Table 2 compare RH0 values obtained by calculating
other physical properties. For this example, moisture RH0 values for mixtures from the Ross equation and
uptake onto cromolyn sodium was correlated with those obtained experimentally. Agreement is very
expansion of the lattice in the b crystallographic direc- good, especially considering the high levels of dissolved
tion, which was shown to be reversible on dehydration. solute(s) that are attained (i.e., estimated as high as 50
A thorough understanding of the hydration profile molal for the choline bromide/tetrabutylammonium
for a solid forming a crystal hydrate is important bromide system).[43]
for several reasons. First, because an anhydrate and
hydrate(s) are distinct thermodynamic species, they
will have different physicochemical properties (e.g., Table 1 RH0 values for single component systems at 25
C
solubility) that may affect dissolution or bioavail- Compound RH0 References
ability. Second, a desired hydrate species can be formed Potassium chloride 84 [41]
and used (and retained) simply by controlling the Potassium bromide 81 [41]
established environmental conditions. Third, because
Potassium iodide 68 [41]
significant quantities of water can be sorbed/liberated
as a hydrate becomes hydrated/ dehydrated, the physi- Sodium chloride 75 [41]
cochemical properties of the immediate system (includ- Choline iodide 72 [41]
ing other nearby solids) can be markedly affected. Choline bromide 41 [41]
Choline chloride 23 [41]
Tetrabutylammonium bromide 61 [41]
The Critical Relative Humidity, RH0
Potassium acetate 23 [31]
Knowledge of RH0 for each component in a formu- Potassium carbonate 43 [31]
lation, and for the entire system, is extremely impor- Sucrose 84 [41]
tant for predicting relative humidities where gross Fructose 64 [41]
physical changes of the system are expected due to Glucose 87 [41]
dissolution of water-soluble components. The value Sodium salicylate 79 [39]
of RH0, as a colligative property, is significantly influ-
Sodium benzoate 88 [39]
enced by the number of species in solution. As a rule of
Water–Zeta

thumb, two general comments can be made. First, Salicylic acid >99 [53]
compounds exhibiting poor water solubility typically Benzoic acid >99 [53]
have RH0 values well above 95% relative humidity. Malic acid 78 [53]
Second, as solubility increases, RH0 decreases. Since Tartaric acid 93 [53]
non-idealities are introduced as solutions become more Fumaric acid 98 [53]
and more concentrated, it is not usually possible to use
Succinic acid 95 [53]
dilute solution models (e.g., Raoult’s law) to predict
 4058 Water Sorption of Drugs and Dosage Forms

Table 2 RH0 values for single component systems at 25
C

Mixture RH0 calculated Experiment
Sodium cloride–potassium bromide 61 64
Potassium chloride–sodium chloride 64 67
Potassium chloride–potassium bromide 68 73
Sucrose–potassium bromide 68 66
Sucrose–dextrose monohydrate 69 68
Sucrose–sodium chloride–potassium bromide 51 57
Choline bromide–potassium bromide 33 40
Tetrabutylammonium bromide–potassium bromide 49 57
Tetrabutylammonium bromide–choline bromide 25 34

The Kinetics of Deliquescence above RH0 crystalline substances below their critical relative humi-
dities. Typical substances of pharmaceutical interest
Initial work by Edgar and Swan,[55] Adams and in this class of solids include celluloses, starches,
Merz,[56] Prideaux,[57] Morkowitz and Boryta,[58] and poly(vinylpyrrolidone), gelatin, and some lyophilized
Carstensen[1] suggested that the rate of moisture proteins. Although some of these substances exhibit
uptake onto water-soluble solids above RH0 should partially crystalline character, they generally contain
depend on the difference between the partial pressure significant fractions of amorphous material and, thus,
of water in the environment and that of the partial fall into this class of substances. A typical isotherm
pressure of water above a saturated solution of the for microcrystalline cellulose is shown in Fig. 4. Note
water-soluble substance, the temperature, the exposed the significant amounts of water that are sorbed over
surface area of the solid, the velocity of movement of the entire relative humidity range and that both the
the moist air, and a specific reaction constant that is sorption and desorption isotherms are characterized
characteristic of the individual solid. by the classical sigmoidal shape often observed with
Van Campen, Amidon, and Zografi[41] developed the physical adsorption of gases. Also apparent is
models describing the rate of moisture uptake above the hysteresis between the sorption and desorption
RH0 that consider both the mass transport of water portions of the isotherm (i.e., the amount of water
to the solid substance and the heat transfer away from associated with the solid is greater for the desorption
the surface. For the special case of an environment isotherm than the sorption isotherm for a given
consisting of pure water (i.e., initial vacuum conditions), relative pressure). This is typical for these types of
the Van Campen, Amidon, and Zografi model is greatly materials and is generally attributed to either kinetic
simplified because vapor diffusion need not be con- effects or to a change in the polymer chain confor-
sidered. Here, only the rate at which heat is transported mation caused by plasticization effects of sorbed
away from the surface is assumed to be an important water.[19,59–62]
factor in limiting the sorption rate, W0 . For this special Fig. 4 also shows the excellent fit to the GAB
case, an expression was derived to express the rate of equation Eq. (10) of the sorption and desorption iso-
moisture uptake solely as a function of RH1, the relative therms for microcrystalline cellulose. In this regard,
humidity of the environment, and RH0. this equation offers considerable practical utility in
This model was shown to be applicable for describ- fitting isotherms for these types of materials over the
ing moisture uptake kinetics (in vacuum) above RH0 entire relative humidity range, especially in contrast
for single component systems of alkali halides, sugars, to the BET equation, which usually only fits uptake
and choline salts.[41] The model later was extended to data up to about 40% relative humidity. As previously
consider the moisture uptake kinetics above RH0 for mentioned, however, this does not in itself confirm the
multicomponent systems of these substances.[43] validity of the GAB model for describing moisture
sorption data on these materials. Rather, independent
Water–Zeta

confirmation of the physical meaning is necessary.

WATER SORPTION BY AMORPHOUS SOLIDS Considerable physical insight has been gained into
the primary binding mechanism of water onto starches
Isotherm Analyses at Ambient Temperatures and celluloses from isotherm analyses that yield values
for Wm [Eqs. (5), (7), and (10)]. This is illustrated in
The amount of moisture sorbed by amorphous solids is Table 3, which gives Wm values for three types of
typically much greater than that sorbed by non-hydrating starches.[61] Table 3 shows that the values for Wm are
Water Sorption of Drugs and Dosage Forms 4059

0.25 be precisely the case, it is not surprising that the values

measured for Wm are slightly less than 0.11 g/g.
Zografi et al.[61,64] have extended this analysis to the
sorption of water by various celluloses. For celluloses,
0.20
corrections are necessary because only the amorphous
regions of cellulose take up water vapor. Table 4 shows
the Wm values obtained from isotherm analyses of sev-
eral cellulosic materials after accounting for the degree
W (G water / G solid)

of crystallinity. As expected, celluloses with different

0.15 degrees of crystallinity exhibit different values of Wm
without correction for crystallinity, and all are consider-
ably less than that for the starches. When corrected,
however, for the degree of crystallinity, all of the values
0.10 are in reasonable agreement with each other and with
the Wm values obtained for the starches. Especially
interesting are the results in Table 4 for microcrystalline
cellulose samples having different degrees of crystal-
linity due to grinding.[66] These results suggest that
0.05
a similar mechanism of water uptake is occurring in
starches and the non-crystalline regions of celluloses.
Similar analyses of moisture uptake data available
in the literature for other cellulose and starch deriva-
0.00
tives used as pharmaceutical excipients are presented
0.0 0.2 0.4 0.6 0.8 1.0 in Table 5. Considering the uncertainties associated
Relative pressure with the estimated moisture uptake values from pub-
Fig. 4 Water vapor sorption (&) and desorption (`) iso- lished graphs, the values of Wm are all quite consistent
therms for microcrystalline cellulose at 20
C. Solid line: GAB with each other and with a stoichiometry of one water
fit to sorption data; Dashed line: GAB fit to desorption data. molecule per anhydroglucose unit. It is interesting to
note that the two samples derived from cellulose,
sodium carboxymethylcellulose and sodium croscar-
quite constant despite significant morphological differ- mellose, did not require any correction for degree of
ences between the various starches. One value of Wm, crystallinity to conform to close to a 1 : 1 stoichiometry.
taken from a fit of desorption data, appears to be It appears quite likely, therefore, that the change in
slightly higher than values obtained from sorption chemical structure and the processing of these materials
data. This might be expected if the availability of essentially eliminates the crystallinity of cellulose.
primary sorption sites had been increased by previous The preceding analysis suggests that water, indeed,
exposure to elevated relative humidities, with sub- penetrates throughout the amorphous regions of these
sequent increased levels of water sorption. As shown materials and undergoes a specific interaction with
by Van den Berg et al.,[19,59,60] these values of Wm are available sorption sites, most likely the available
all close to the value of 0.11 g of water per g of starch, hydroxyl groups on the anhydroglucose units. Differ-
calculated by assuming that one water molecule sorbs ential heat of sorption results for various starches[19,63]
per anhydroglucose unit. Because this calculation
assumes that all anhydroglucose units are available
for primary binding, and because this is not likely to Table 4 Wm values for various celluloses obtained from
BET analysis of moisture uptake isotherms corrected for
degree of crystallinity
Table 3 Wm values for various starches obtained from BET
analysis of moisture uptake isotherms Cellulose Crystallinity % Wm Corr (g/g) References

Starch Wm (g/g) References Cotton 70 0.093 [65]

a Cellophane 40 0.098 [65]
Water–Zeta

Corn 0.095 [39]

Corn 0.083 [63] MCC 63 0.095 [66]

Potato 0.085 [19] MCCa 49 0.076 [66]

a
Wheat 0.080 [19] MCC 38 0.107 [66]
a
This value is taken from the desorption isotherm. Others are from MCCa 0 0.086 [66]
a
sorption isotherms. MCC Ground in a ball mill.
(From Ref.[61].) (From Refs.[61,66].)
 4060 Water Sorption of Drugs and Dosage Forms

Table 5 Wm values for various pharmaceutical excipients decreases as the temperature increases, reflective of
obtained from BET analysis of moisture uptake isotherms an overall exothermic process, normally expected with
Excipient Wm (g/g) References vapor adsorption processes. Such behavior has been
Starch 1500 0.074 [63]
observed with cellulose,[85] starch,[19] poly(vinylpyrroli-
done),[13] and poly(methyl methacrylate).[86] In such
Sodium starch glycolate 0.081 [67]
cases it is often assumed that the dominant factor is
(ExplotabÕ)
the negative heat of absorption arising from the change
Sodium starch glycolate 0.092 [67] in the extent of water binding. The process, however,
(PrimogelÕ)
is made much more complex than this because of the
Crosslinked dextrose (CLD-2) 0.098 [68] changing morphology of the solid and, hence, an
Croscarmellose, sodium 0.094 [68] entropy change as well. The complexity of the effects
(Ac-Di-SolÕ) of temperature on water vapor absorption and the
Sodium carboxymethylcellulose 0.103 [69] possible links to the plasticizing effects of water may
(From Ref.[61].) be observed in the work of Oksanen and Zografi,[13]
who have reported that the Wm values for poly(vinyl-
pyrrolidone) over the temperature range of 40–60
C
decrease by a factor of three, suggesting that Wm does
and cellulose[10,68] support this model. Data gathered not reflect the absolute number of available binding sites
indicates that there is a specific water–solid interaction on the polymer for directly ‘‘bound’’ water. Rather, Wm
out to a moisture content of at least the equivalent of appears to be related to W(Tg ¼ T the amount of water
3 times Wm. The water present in this system appears sorbed that will reduce the glass transition temperature,
to exist in a more structured state (i.e., reduced mobility) Tg, to the temperature of the sample, as the ratio of
than bulk water over this range. Interestingly, the heats W(Tg ¼ T)/Wm remains nearly constant at 3.0 over
of sorption exhibit discrete breaks corresponding to the entire temperature range.
stoichiometries of one and two water molecules per In summary, it is clear that water absorbs into
anhydroglucose unit. Some differential heat of sorption amorphous polymers to a significant extent. Inter-
results are nearly constant over the Wm range, suggesting action of water molecules with ‘‘available’’ sorption
that binding is homogeneous over this range.[10,19] This sites likely occurs via hydrogen bonding such that
is, however, not always the case. This is borne out by the mobility of the sorbed water is reduced and the
heat of vaporization data reported by Etzler and thermodynamic state of water is significantly altered
Conners on cellulose samples.[70] As measured using relative to bulk water. Yet accessibility of the water
simultaneous DSC/TGA, the heat of desorption of to all potential sorption sites appears to be dependent
water from the matrix continuously increases once the on the previous history and physicochemical properties
moisture content is less than approximately 0.5 g of the solid. In this regard, the water–solid interaction
water/g cellulose (about 3 times Wm). These results in amorphous polymer systems is a dynamic relation-
suggest that the sorbed water exists in multiple ‘‘states’’ ship depending quite strongly on water activity and
that are energetically distinct. temperature.
Other supportive evidence for a specific water–solid
interaction is available from thermal studies showing
the amount of non-freezeable water,[71–73] nuclear mag- The Meaning of Specific Surface
netic resonance,[39,74–80] and diffusion studies.[81,82] The Areas Calculated from Water
evidence is less clear, however, concerning whether Absorption Studies
there is distinct binding of water to sorption sites with
discrete energy levels or whether there is a continuum Simply calculating specific surface areas from the Wm
of states where water interacts to a lesser extent with values in Tables 3–5 leads to ‘‘apparent’’ specific
increasing amount sorbed.[61,83] In any event, it is clear surface areas of approximately 300–500 m2/g.[61,64]
that sorbed water behaves with a considerable degree Specific surface areas obtained from similar analyses
of mobility, and hence, questions the use of the term of non-polar gas (nitrogen or krypton) adsorption
‘‘bound water.’’[61,84] studies, however, are typically in the range of 1 m2/g,
Water–Zeta

independent of sample pretreatment.

Interestingly, the ball milling studies of microcrys-
Isotherm Analyses as a Function talline cellulose by Nakai (Table 4)[66] have shown that
of Temperature the Wm values obtained from water sorption studies
increase to a much greater extent than the increase in
Generally speaking, the absorption of water into surface area because of comminution of the sample.
amorphous solids as a function of relative humidity In fact, as discussed earlier, moisture sorption was
Water Sorption of Drugs and Dosage Forms 4061

shown to be proportional to the amount of amorphous 0.6

character, suggesting that water is absorbed through-
out the amorphous regions of this substance. In this
regard, artifactual specific surface areas are obtained
if calculated from water absorption data[64] for these 0.5
types of substances.

W (G water / G dry PVP)

0.4
The Role of Water as a Plasticizer

Absorption of significant amounts of water into the

internal structure of a solid has been shown to influ- 0.3
ence the properties of the solid. This is apparent, for
example, in the hysteresis observed between the sorp-
tion and desorption isotherms in Fig. 4. This phenom-
enon becomes exaggerated to a greater extent for 0.2
materials that consist of higher proportions of
amorphous material. Levine and Slade[14,87] have
demonstrated that water, with a very low glass tran-
sition temperature, can act as a plasticizer, thereby 0.1
lowering the glass transition temperature, Tg, of
amorphous polymers. Similar behavior is observed in
amorphous low molecular weight solids. Recognizing
0.0
that the viscoelastic properties of the solid are
altered significantly above Tg (rubbery state) relative 0.0 0.2 0.4 0.6 0.8 1.0
to below Tg (glass or vitreous state), it is likely Relative pressure
that the solid will undergo changes of its physical Fig. 5 Water vapor sorption isotherms for poly(vinylpyrro-
properties at distinct moisture contents and defined
temperatures as a result of this phenomenon.[13,14]
lidone) at 60
C (þ); 30
C (); 20
C (Z); 40
C (`). A 
represents the calculated water contents necessary to depress
Oksanen and Zografi[13] have shown with poly(vinyl- Tg to the temperature of the isotherms. (Data were taken
pyrrolidone) that the moisture content at which the from Oksanen and Zografi[76,78].)
moisture sorption isotherm begins to increase signifi-
cantly correlates very well with the moisture content
that will reduce Tg to the temperature of the isotherm. simplified Gordon–Taylor/Kelly–Bueche equation:[90]
This is illustrated in Fig. 5, which shows water absorp-  
tion isotherms for poly(vinylpyrrolidone) over the 1 W1 W2
¼ þ ð16Þ
temperature range of 40 to 60
C.[13,88] Clearly, the Tg ðmixÞ Tg1 Tg2
inflection point at which the isotherm begins to turn
markedly upward shifts to a higher moisture content where W1 and W2 are the weight fractions of compo-
as the temperature is reduced. To illustrate this, note nents with Tg values Tg1 and Tg2. The results clearly
that the moisture content (0.674 g/g) necessary to indicate that the viscoelastic properties of amorphous
reduce Tg to 40
C has not been attained yet in materials can undergo significant changes as the solid
Fig. 5 and the isotherm appears quite linear over the transitions from the glassy to rubbery state. Further-
relative humidity range shown. For further clarity, more, these changes can occur due to elevation of
the moisture contents [W(Tg ¼ T)] corresponding to temperature at fixed moisture content or to an increase
Tg at 60, 30, 20
C, and 40
C were shown to be in moisture content at constant temperature or to a
about 0.205, 0.313, 0.553, and 0.674 g/g, respectively. combination of these effects. For dry amorphous sub-
Oksanen and Zografi[13] reported that cellulose and stances, molecular mobility of the solid begins to be
elastin (a protein) exhibit similar relationships, where significantly enhanced relative to the glassy state as
Water–Zeta

the glass to rubber transitions correspond to the low as 50
C below Tg.[91] Similar increases in molecular
upward inflections in their respective isotherms. mobility due to the plasticizing effects of absorbed
Subsequent studies by Hancock and Zografi[89] water suggest the need to maintain amorphous systems
demonstrated that the glass transition temperatures at least 50
C below the system glass transition
for PVP, hydroxypropyl methylcellulose, and poly temperature to avoid physical, chemical, and/or mech-
(methyl methacrylate) were linearly depressed by the anical property changes over the product shelf life.
weight fraction of sorbed water, according to the Some properties that are likely to be affected include
 4062 Water Sorption of Drugs and Dosage Forms

tablet compaction,[92,93] gelatin capsule brittleness,[14,94] surface area as relative humidity increased, due to the
collapse of lyophilized amorphous powders,[88,95,96] consequent recrystallization of the disordered surface
protein stability,[97,98] and the stability of low molecular material.[48] Fukuoka, Makita, and Yamamura[116]
weight, moisture sensitive drugs mixed with amorphous have demonstrated that a variety of pharmaceutical
polymeric substances.[99] substances indeed can be made amorphous and,
furthermore, exhibit glass transition temperatures over
a range from 243 to 354 K. For example, aspirin, pro-
WATER SORPTION BY PHARMACEUTICAL gesterone, phenobarbital, and sulfadimethoxine exhibit
SOLIDS SUBJECTED TO PROCESSING Tg values of 243, 279, 321, and 339 K, respectively.
Similar to amorphous polymeric systems, low
Understanding the mechanisms of moisture sorption molecular weight amorphous substances also exhibit
by solids existing in either the crystalline or amorphous a reduction in Tg as moisture content increases,[117]
states allows a conceptual estimation of critical points thereby leading to favorable conditions for recrystalli-
where major changes in physical or chemical properties zation to occur. Indeed, low molecular weight amorph-
occur (e.g., RH0, a crystal hydration relative humidity, ous solids possess sufficient molecular mobility well
glass transition temperature). Processing (i.e., milling, below Tg.[118] In some systems with multiple solid states
spray drying, compaction, lyophilization, etc.) of possible, the water activity/moisture content can
pharmaceutical solids, however, often induces at least influence the crystallized form of the solid.[119] Unfor-
partial conversion of most substances to a high energy tunately, recrystallization of non-hydrating, low
form.[100–107] Such local disorder has been associated molecular weight amorphous systems can lead to the
with enhanced chemical reactivity[101–108] and increased liberation of significant amounts of water to the head-
solubility[15] relative to the thermodynamically favored space.[15,48,103,104,120] Such ‘‘moisture dumping’’ can
crystalline state. These regions have been referred to as have additional impact on the physical, chemical, and
‘‘hot’’ spots of the bulk solid and, when present, leave mechanical properties of the system.[17,48,100]
the solid in an ‘‘activated state’’.[15,100–107,109–113] To illustrate this more quantitatively, consider the
This non-homogeneity that exists in processed hypothetical sucrose example discussed by Ahlneck
solids complicates the study of moisture sorption and Zografi.[100] Assuming that all the sorbed water
phenomena in these materials, as more than one mech- is taken up by the amorphous portion of material,
anism of uptake must be considered. This is especially 0.1% total moisture would correspond to approxi-
difficult, and often frustrating, for cases in which only mately 20%, 10%, 4%, and 2% moisture content in
a small amount of amorphous material is present, as the amorphous material, respectively, for 0.5%, 1%,
the experimental techniques required to complete these 2.5%, and 5% of amorphous solid. The glass transition
analyses are labor intensive.[114,115] Yet, relatively low temperatures for the amorphous portions of these sys-
percentages of amorphous material can absorb con- tems range from 9 to 49
C, respectively.[87,100] Hence,
siderable amounts of water into their structure, with significant changes in the solid state properties are
these regions undergoing considerable change and a expected at room temperature if relatively small
consequent effect on the overall properties of the bulk amounts of amorphous material (i.e.,<1%) are initially
substance.[100] This is especially important for low mol- present. This example illustrates that even for low
ecular weight substances that have the ability to readily moisture content materials, significant changes can
recrystallize due to their overall greater mobility rela- occur in localized amorphous regions of a solid, which
tive to higher molecular weight polymeric materials. may affect properties of the material influenced by
This has been demonstrated for sodium chloride and molecular mobility.[100]
sodium salicylate ground for 15 min in a mortar and Inhibition of events resulting from increased mol-
pestle.[48] Whereas recrystallized materials exhibited ecular mobility due to increased moisture absorption
no change in specific surface areas with increasing rela- and a subsequent reduction in Tg can be accomplished
tive humidities, the ground samples exhibited signifi- by formulating such materials with amorphous sub-
cant reductions in specific surface areas as relative stances of higher Tg. The net effect is to increase
humidities were increased. Fig. 2 illustrates the differ- molecular interaction and raise the system Tg to a level
ing moisture uptake profiles for the recrystallized and where molecular mobility is again sufficiently low
Water–Zeta

ground sodium chloride samples, normalized for spe- (high viscosity) such that the undesired property
cific surface area.[48] Whereas the ground material changes do not occur.[107,121,122]
sorbed significantly more water at lower relative The recrystallization of amorphous low molecular
humidities than the recrystallized sample, the recrystal- weight systems can be convoluted by the impact of
lized material sorbed greater amounts at higher relative structural changes on the material. For example, spray
humidities. This relative reduction in sorption capacity drying a-lactose monohydrate typically produces a
of the ground sample is attributed to a reduction in material that is completely amorphous as determined
Water Sorption of Drugs and Dosage Forms 4063

by powder X-ray diffraction. However, lactose under- moisture contents and dry weights, headspace volume,
goes anomeric rotation in solution, causing a change and temperature. Final moisture contents for each
in the fundamental structure of the molecule. The solid can then easily be estimated from the isotherms
impact of this structural change on the uptake and for the respective solids.
equilibration of water to the amorphous lactose is sig- The SDMT model has practical utility in aiding the
nificant. As expected, an increase in moisture content rational optimization of the initial moisture contents of
will result in the suppression of the glass transition individual components in a system to attain the final
temperature to the point where instantaneous crystalli- desired relative humidity. Practical applications to date
zation occurs. However, the material produced is non- have included adjustment of the initial formulation
uniform as there are two different forms of lactose LODs prior to capsule filling to avoid gelatin capsule
present, one of which is anhydrous in the crystalline brittleness,[94,127] selecting the appropriate formulation
state. Furthermore, at elevated humidity, the crystal- moisture content and amount of desiccant to maintain
line b-anhydrate can undergo structural change in the the relative humidity inside a container below a defined
solid state and produce the a-monohydrate. Evalu- value,[128] and selection of appropriate dry powder
ation of data gathered needs to be completed in the inhaler design and packaging conditions for optimal
context of a careful characterization of the possible stability.[129]
solid forms.[16,17,123–125]

CONCLUSIONS
TRANSFER OF WATER BETWEEN SOLID
COMPONENTS VIA THE HEADSPACE Moisture is present in all solid pharmaceutical drugs
and dosage forms and in most processing techniques.
Combining solids that have previously been equili- Understanding where the water resides, its state, and
brated at different relative humidities results in a sys- the manner in which it affects the properties of individ-
tem that is thermodynamically unstable because there ual materials, their mixtures, and ultimately, final pro-
will be a tendency for moisture to distribute in the sys- duct performance and integrity are essential for the
tem so that a single relative humidity is attained in the developmental scientist to better understand the role
headspace. As shown in Fig. 6, moisture will desorb of water in a particular system. Especially important
into the headspace from the component initially equili- are the kinetics of moisture uptake or loss, ‘‘equilib-
brated at a higher relative humidity and sorb to the rium’’ uptake values as a function of relative humidity,
component initially equilibrated at a lower relative whether the water resides externally or is absorbed into
humidity. This process will continue until both solids the material, its degree of binding with the solid, and
have equilibrated at the final relative humidity. The the tendency for water to redistribute in a system con-
final relative humidity can be predicted a priori by sisting of more than one solid. Although water–solid
the sorption–desorption moisture transfer model interaction(s) can be extremely complex in pharmaceu-
(SDMT) model[126] if one has moisture uptake iso- tical systems, application of these fundamental con-
therms for each of the solid components, their initial cepts to product development can greatly aid in
understanding the role of moisture in affecting the
physicochemical properties of solid materials.
A B

RA RB Rf
VA VB VT ACKNOWLEDGMENTS

The authors thank Professor George Zografi, Dr.

Cindy Oksanen, and Dr. Frank Etzler for their techni-
cal contributions to this article and Ms. Linda Schwei-
A B A B kardt for her graphical support.
Fig. 6 Schematic representation of moisture transfer
Water–Zeta

between solid components, A and B, respectively. (A) Head-

spaces isolated from one another; (B) Headspaces allowed to
equilibrate. RA and RB are initial relative humidities above A
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Ref.[126].) dynamics; McGraw-Hill: New York, 1961.
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Water–Zeta
 Waxes
Roland A. Bodmeier
College of Pharmacy, Freie Universität Berlin, Berlin, Germany

INTRODUCTION concerns with animal-derived products, spermaceti has

been replaced with other natural or synthetic products.
The term wax generally refers to a substance that is a The most commonly used insect wax is beeswax. It
plastic solid at room temperature and a liquid of low is obtained from the honeycomb of the bee. White
viscosity above its melting point. Strictly speaking, a and yellow beeswax are GRAS-listed and consist of
wax is chemically defined as an ester of a monohydric mixtures of various esters of straight chain monohydric
long chain fatty alcohol and a long chain fatty acid. alcohols with even number carbon chains (C24–C36)
However, generally the term wax has been applied to esterified with straight chain fatty acids. The major
a broad group of chemically heterogeneous materials. ester is myricyl palmitate. Beeswax also contains free
Waxes usually contain a wide variety of materials acids and carbohydrates. White wax is obtained
including glycerides, fatty alcohols, fatty acids, and through bleaching of yellow wax with oxidizing agents
their esters. In the pharmaceutical literature, the terms or with sunlight. The National Formulary 18 (NF18)[8]
waxes, fats, or lipids have often been used inter- specifications list a melting range of 62–65
C, an acid
changeably and no consistent terminology has been value of 17–24, and an ester value of 72–79. It is prac-
established. They have in common their lipophilic tically insoluble in water, sparingly soluble in ethanol,
character and their insolubility in water and solubility and soluble in chloroform and various oils. Beeswax is
in non-polar solvents. Besides natural materials, many used as a stiffening agent in topical preparations, as a
semisynthetic products such as fatty acids or alcohols stabilizer of w/o-emulsions, and as a polishing agent in
or surfactants are derived from lipids. sugar coating.
Waxes have been used by the pharmaceutical indus- Carnauba wax is plant-derived and is obtained from
try for many years. Their applications in semisolid pre- the carnauba palm tree, indigeneous to Brazil. The wax
parations, including ointments, creams, or lotions, and is obtained from the surface of dried leaves. It is widely
in suppositories are well known and numerous publica- used in food, cosmetic, and pharmaceutical products.
tions exist on this topic. Because of their lipophilic It consists of a complex mixture of high-molecular-
properties, waxes have been used in sustained-release weight esters of acids and hydroxyacids. Carnauba
single or multiple unit solid dosage forms. This article wax is very hard and brittle, and has a high melting
reviews the different uses of waxes as sustained-release point. The NF18 specifications list a melting range of
carrier or coating materials. 81–86
C, an acid value of 2–7, and a saponification
value of 78–95. It is insoluble in water, slightly soluble
in boiling ethanol, and soluble in warm chloroform.
Besides the sustained-release applications described later,
WAXES IN PHARMACEUTICAL it is used as a polishing agent in sugar coating because of
DOSAGE FORMS its high gloss, and in topical preparations. Other, less
used vegetable-derived waxes include candelilla wax
Waxes are obtained from various sources and are gen- and castor wax.
erally classified into animal, insect, vegetable, mineral, Hydrogenated vegetable oils are prepared by hydro-
and synthetic waxes.[1–7] genation of refined vegetable oils. Hydrogenated veg-
The most familar animal wax is probably lanolin, etable oil consists of mixtures of triglycerides, with
which is obtained from the wool of the sheep. It con- two types being defined in the USP23. Type II includes
Water–Zeta

sists primarily of esters of C18–C26 alcohols and fatty partially hydrogenated vegetable oils and has a lower
acids, sterols (cholesterol), and terpene alcohols. It is melting range and a higher iodine value than Type I.
frequently used in topical preparations. Until recently, Type I melts in the range of 57–70
C and has iodine
spermaceti was another commonly used animal wax. value of 0–5, while Type II has a melting range of
Spermaceti is obtained through the precipitation of 20–50
C and an iodine value of 55–80. They are used
the head oil from the sperm whale on cooling. It as lubricants, as sustained-release matrix materials, as
consists primarily of cetyl palmitate. Because of public viscosity modifiers in semisolid formulations, to enhance
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-100001728
4066 Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved.
Waxes 4067

the solidification of suppositories, and to minimize the The congealing point of a wax is the temperature at
sedimentation of dispersed drug. which the molten wax stops to flow upon cooling. Ther-
Two commonly used mineral-derived waxes are mal methods such as differential scanning calorimetry
petroleum wax, which is microcrystalline, and paraffin (DSC) are widely used to characterize the heating and
wax, which is crystalline. They are both obtained from cooling profiles of waxes in a qualitative and quantitat-
petroleum: the quality and quantity of the wax ive manner. Potential polymorphic transitions and
depends on the source of the crude oil and the refining recrystallization during processing can be simulated by
process. Microcrystalline wax (petroleum ceresin or running different temperature profiles.
wax) consists of straight chain and branched saturated The contraction of suppository bases during cooling
alkanes with a chain length range C41–C57. The NF18 within the mold is a well-described phenomenon. The
specifications list a melting range of 54–102
C; it comes expansion or contraction of waxes is also important
in plastic and hard grades. It is insoluble in water, during the processing of wax melts, for example, dur-
slightly soluble in ethanol, and soluble in chloroform. ing the preparation of microparticles by spray congeal-
Besides its use as a sustained–release carrier, it is used ing, hot-melt coating, or hot-melt filling of hard gelatin
as a stiffening agent in topical preparations. Because capsules. The dilatation of waxes or thermal expansion
of its high viscosity and melting point, it increases during the transition from the solid to the liquid state
the consistency of creams and ointments. Paraffin can be measured with a dilatometer. The hardness of a
wax (hard paraffin) is a mixture of solid straight chain wax is measured with a penetration test, whereby the
alkanes. It is used in ointments or creams as a base or depth of penetration of a needle under a given weight
stiffening agent. It congeals between 47 and 65
C. is measured, preferably at different temperatures. The
Various grades with different melting ranges are avail- viscosity of the molten wax is an important parameter,
able. It is insoluble in acetone, ethanol, and water and especially for processes such as hot-melt coating or
soluble in chloroform and most warmed fixed oils. spray congealing, where wax melts are processed. In
Low-molecular-weight polyethylenes (MW < 10,000) an ASTM monograph (D 88), the time that a certain
have wax-like properties and are used in topical pre- quantity of molten wax requires to flow through an
parations, for example, as gelling agents in Plastibase. orifice of specified dimensions is measured.
The color of the wax will affect the color of the
finished product. A Lovibond Tintometer is often used
CHARACTERIZATION for color measurements, whereby the color of the raw
material is compared against a series of colored stan-
Because the harvesting of vegetable or insect waxes is dard glasses, under a standard light source. The color
often from wild, non-cultivated sources and because of the solidified wax of the same sample may be differ-
of their complex composition, it is important to char- ent depending on the amount of occluded air, the rate
acterize the chemical and physical properties of the of cooling, or surface finish. Therefore, the color of
waxes.[1–6] The composition of natural materials often many waxes is best measured in the molten state.
varies with location, weather, season of harvesting, Two ASTM color standards are used to measure
and age. A good quality control of the raw materials dark-brown to off-white color and off-white to pure
is of upmost importance in order to obtain pharmaceu- white. The refractive index and the specific gravity
tical products of high quality. are other parameters often determined.
The chemical methods to characterize waxes include The structural and physical properties, in particular,
the determination of the acid, saponification, iodine, the solid and liquid state behavior of lipids, and the
hydroxyl, and peroxide values. Various tests, often optical and spectral characteristics of waxes have been
yielding different values, are available to measure the described in detail.[4]
melting point of waxes. Since waxes are non-
homogeneous in chemical composition, a melting range
rather than a clear melting point is most observed. The
melting point of glycerides generally increases with PHARMACEUTICAL APPLICATIONS
increasing hydroxyl number, decreasing degree of unsa-
turation, and increasing molecular weight of the fatty Waxes in Matrix-Type Drug Delivery Systems
Water–Zeta

acid. The melting point of many waxes can be determ-

ined with capillary tubes. The slip point is defined as The incorporation of drugs into inert matrices is a
the temperature at which a column of the testing popular approach to prolong the drug release.
material starts raising in an open-ended capillary tube, Sustained-release wax matrix drug delivery systems
which is dipped in water filled in a beaker and heated include wax granules or beads prepared by granulation
under specific conditions. The drop-point test can be or extrusion/spheronization, tablets, and wax-filled
used; however, it is not reliable for more viscous waxes. hard gelatin capsules.
 4068 Waxes

Wax Granules and Beads release. The beads were subsequently compressed into
tablets, which had slower release rates when compared
Drug-containing wax granules were prepared by melt to the release from the corresponding beads. While
congealing, by congealing in chloroform, by granu- non-treated compacts mainly disintegrated into the
lation, and by aqueous dispersion.[9–11] In the congeal- beads, thermal treatment of the compacts resulted in
ing method, the drug was suspended in the molten wax. non-disintegrating matrices.
This suspension was cooled gradually while stirring Diclofenac sodium–carnauba wax matrix granules
until a solid mass formed, which was then comminuted containing various release-controlling agents such as
into granules. Granules made by congealing in chloro- hydroxyporpylcellulose, Eudragit L-100, or NaCl were
form were prepared similarily, the drug being sus- prepared with a twin-screw extruder and evaluated in
pended in a chloroformic solution of the wax. This vitro and in vivo.[14,15] The drug release depended
mixture was agitated until the solvent evaporated, strongly on the composition of the granules: the mech-
and was then comminuted into granules. In the granu- anical strengths of the wetted granules was high, and a
lation method, the powdered wax and drug were good correlation was obtained between in vitro dissol-
granulated with chloroform. In the last method, heated ution parameter and in vivo parameters.
water was given to the molten drug–wax mixture until
phase inversion occurred. The emulsion was cooled and
the particles were separated from the aqueous phase by Wax Matrix Tablets
filtration. The melt-congealing method gave the largest
retardation in drug release, probably because of the den- Wax matrix tablets are prepared either by compression
ser structure of the granules when compared to granules of the wax granules or beads described in the previous
prepared by the other methods. section, or, with the higher melting waxes available in
Sustained-release nitrofurantoin granules contain- powder form, by direct compression of powder blends.
ing stearic acid and glycerol monostearate as matrix When compared with polymeric matrix materials, the
materials were prepared by either a fusion, solvent amount of waxes within the tablet is limited because
evaporation, or melt granulation technique.[12] Various of fusion and sticking to the punches at higher wax
channelling agents including Aerosil, Avicel, dibasic concentrations. This problem could possibly be over-
calcium phosphate dihydrate (Emcompress), and come by compression at lower temperatures.
sodium chloride were investigated in order to increase Matrix tablets containing ephedrine hydrochloride
the drug release. In the fusion method, the lipid was and hydrogenated castor oil were prepared either by
melted and the drug was added to the melt. After cool- compression of a physical mixture or by compression
ing, the congealed mass was granulated through stan- of a congealed melt.[16–18] In the second method, the
dard sieves. In the solvent evaporation method, the drug was added to the molten hydrogenated castor
drug and the wax carriers were dissolved in dimethyl oil at 100
C, this molten mass was poured onto a glass
formamide. The solution was cast and the solvent plate and congealed, comminuted and then compressed
was evaporated at 70
C. The resulting mass was granu- into a matrix. A surfactant, 0.1% alkyltrimethylammo-
lated as described previously. In the melt granulation nium bromide, was added to the dissolution medium to
method, the drug and carriers were mixed at high enhance the wetting of the wax matrix. The drug
speeds; the temperature increased by friction and release increased with increasing amounts of drug in
granulation occurred by sintering of the fatty materials the matrix because of an increased porosity, and fol-
near their melting point. The cooled granules were lowed the square root of time relationship (Fig. 1).
crushed and sieved. Sustained release could only be The release was slower from the matrix prepared by
obtained with granules prepared by the fusion method. the melt method. This was attributed to a higher
The solvent evaporation method and, surprisingly, melt tortuosity and lower porosity of the melt matrix when
granulation resulted in granules with a fast drug release. compared to the matrix prepared from the physical
Drug-containing beads were prepared by extrusion/ mixture. Increasing the pressure decreased the release
spheronization of powder blends of 10% drug (chlor- rate with both preparation processes; however, the
pheniramine maleate or acetaminophen), 60% Avicel effect was much more pronounced for the matrix
PH-101, and 30% wax.[13] Thermally treating the prepared from the physical mixture. The processing
Water–Zeta

beads at 80
C for 30 min resulted in the melting and method also altered the mechanism and rate of release;
recongealing of the wax within the beads and in a a matrix diffusion mechanism was dominant with the
decrease in the drug release, probably because of a melt process while a boundary layer diffusion was
densification and redistribution of the wax within effective with matrices prepared by the compression
the beads. The drug release was dependent on the of physical mixtures. In the first case, the drug release
treatment temperature and the level of the wax, with was independent of stirring speed, while it was depen-
an increase in both resulting in a decrease in drug dent in the second case.
Waxes 4069

material for diphenhydramine HCl. The drug and

0.15
Eudragit L100 were mixed, added to the molten wax,
followed by compression of the congealed granules.
The polymer provided an insoluble structure for the
wax. The drug release was significantly retarded with
Q (g/cm2)

0.10 increasing amounts of the anionic polymer.

The cationic drug, diphenhydramine HCl, interacted
with the anionic Eudragit L within the wax matrix
0.05 and formed a complex after water penetration. While
the drug release from tablets prepared from only the
drug and the enteric polymer was highly dependent
on the pH of the dissolution media, the drug release
1 2 3 4 5 was almost independent of pH after the inclusion of
Time (h1/2) carnauba wax in the matrix. Besides the retarding
effect of the wax, at low pH, the release was retarded
Fig. 1 Effect of concentration of ephedrine hydrochloride through the enteric polymer and at high pH, the
on release from matrixes compressed at 7 MPa (48,265 psi). drug formed a less soluble complex with the enteric


( ) 5%; ( ) 10%; (`) 20%; (G) 30%; (&) 40%; (&) 50%.
polymer, thus negating the effect of pH on the drug
(From Ref.[17].)
release.
Matrix minitablets based on starch/microcrystalline
wax mixtures were prepared by melt granulation in a
Sustained-release nifedipine tablets based on Gelu- hot-stage screw extruder, followed by milling and
cires were developed.[19] A drug-PVP coprecipitate compression.[22] This technique was preferred over
was either added to a solution of Gelucire in chloroform the production of pellets.
with the subsequent removal of the solvent or it was Surfactants were incorporated into wax matrices in
added to themolten Gelucire with subsequent cooling order to increase drug release.[23] The drug and surfac-
of the paste. The mixtures were then granulated tant were added to a molten mixture of carnauba wax
through a 0.5 mm sieve and tableted. No difference and stearyl alcohol. Water-insoluble surfactants such
in the release profiles from the two granulates were as glycerol monostearate had no effect on the dissolu-
observed; however, the melting method was preferred tion rate, slightly soluble surfactants such as sodium
because of the absence of organic solvents. Accelerated stearate or dioctyl sodium sulfosuccinate moderately
stability studies on the tablets as a function of tempera- increased the drug release, while polyoxyethylene 23
ture and relative humidity revealed no changes in chemi- lauryl ether significantly increased the drug release.
cal stability; however, the dissolution profiles changed. The drug release occurred via a leaching mechanism;
The changes occurring after storage at high humidities drug diffusion through the matrix did not occur. It
and temperatures were attributed to the formation of was speculated that the surfactant created more chan-
nifedipine microcrystals and to structural changes in nels for the drug to leach into the dissolution medium
the wax vehicle. by increasing the porosity of the matrix.
A combination of ethylcellulose and paraffin wax or While the drug release from wax matrix tablets
hydrogenated castor oil was used as carrier material in followed the square root of time relationship, approxi-
sustained-release aminophylline tablets.[20] The tablet mately zero-order release of ephedrine hydrochloride
granulation was prepared by a wet granulation pro- and procaine hydrochloride could be obtained
cedure with hot ethanol. The tablets were further with multi-layered matrices of hydrogenated castor
coated with Eudragit RL/RS or HPMC/ethyl cellu- oil containing different concentrations of the active
lose. An annealing step at 70
C significantly decreased compound in each layer.[24]
the drug release due to fusion within the tablet core. A bioadhesive lozenge containing cetylpyridinium
Hydrogenated castor oil was superior to paraffin wax. chloride was developed based on a multilayered tab-
Increasing the wax content and decreasing the amount let.[25] One layer contained the bioadhesive polymer,
of ethyl cellulose or increasing the drug content resulted carbopol, and the other layer contained the drug and
Water–Zeta

in faster drug release. a wax (spermaceti or Precirol ATO-5).

In order to overcome the disadvantage of hydrophi-
lic matrices of uncontrollable erosion of the hydrated
polymer gel on the tablet surface, a combined polymer- Wax Implants
wax carrier material was evaluated by Huang et al.[21]
Carnauba wax was combined with the enteric acrylic Besides polymers such as polylactides, waxes have the
polymer, Eudragit L100, and investigated as a carrier potential as biocompatible/biodegradable carriers in
 4070 Waxes

implants. Standard tableting equipment can be used to A Zanasi hard gelatin powder filling capsule machine
prepare wax compacts. (model LZ64) was modified in order to allow the filling
Various lipids including triglycerides (e.g., trilaurin, of molten or thixotropic formulations into hard gelatin
trimyristin, tripalmitin and tristearin) and fatty acids capsules.[33,34] This technique resulted in excellent fill
were evaluated as carrier materials in sustained-release weight uniformity and overcame many of the problems
insulin implants.[26] The drug/wax powder blend was frequently associated with the filling of conventional
compressed into a disc and implanted subcutaneously capsules.
into Wistar rats. Monoglycerides tested eroded too fast The release of liquid or deliquescent drugs (benzo-
and were not suitable. The triglycerides only sustained- natate, nicotinic acid, chloral hydrate, and parametha-
the insulin release briefly. The best sustained-release dione) incorporated into Gelucire bases within hard
properties were obtained with palmitic and stearic gelatin capsules was related to the behavior of the
acids as the carrier materials. carriers in simulated gastric fluid.[35] Gelucires are
The drug release of the model protein, bovine serum semisynthetic glycerides with varying amphiphilic
albumin (BSA), from compressed stearic acid pellets properties and are derived from natural hydrogenated
was investigated as function of drug loading, drug food grade fats and oils. They are characterized by
and carrier particle size, and compression force.[27] At their melting point (range: 33–64
C) and HLB value
low loadings (5%), the drug release increased with (range: 1–13). The drugs were released faster from
increasing BSA particle size irrespective of the particle the Gelucires with the higher HLB value and with
size of stearic acid. At high loading (20%), the drug lower melting points. The Gelucires either dissolved
release was higher with larger stearic acid particles. completely or remained intact but softened.
More BSA was released with increasing BSA particle It is well known that wax-based dosage forms can
size only when the stearic acid particle size was small. experience physical instabilities. Suppository bases
The compression force did not show any effect in the often show an increase in melting point, accompanied
range investigated. In a series of articles, the same by a hardening process. This hardening can result in
research group investigated cholesterol–lecithin implants a reduced release rate, which could also affect the in
as a delivery system for antigens.[28–31] vivo performance. Melt-filled hard gelatin capsules
Labrafil 1944 CS (derivatized vegetable oil)–Precirol were evaluated by DSC, dissolution, and hardness
ATO 5 (glyceryl ester of fatty acids) gels were shown to properties as expressed by a relative penetration.[36]
be biocompatible and biodegradable and resulted in Ketoprofen dissolved in the wax, Gelucire 50/13, and
controlled release of steroids for prolonged periods apparently formed a solid solution at room tempera-
of time in vivo.[32] ture as indicated by the absence of crystalline drug by
DSC and microscopy. The melting point of the wax
increased during storage and was accompanied by a
Hard Gelatin Capsules Filled with Waxes hardening process. However, the release rate increased,
which was attributed to an increased rate of matrix
Waxes are difficult to compress at higher levels. The erosion. The observed in vitro changes did not affect
energy imparted during compaction causes melting of the in vivo performance of the wax matrix.
the waxes, resulting in sticking and picking of the for- As an alternative to hot-melt filling of capsules,
mulation. This necessitates dilution of the drug-wax sssustained-release wax matrices were formed in a
granules with inert fillers; high-dose drugs requiring novel way within hard gelatin capsules through fluidi-
high amounts of wax in order to obtain sustained- zation in a heated air stream within a fluidized bed.[37]
release properties are therefore difficult to formulate A drug–wax powder blend was filled into hard gelatin
into tablets. As an alternative to compressed tablets, capsules, which were then suspended in an upward-
hard gelatin capsules have been liquid-filled with moving, heated airstream and circulated within the
solutions or dispersions of drugs in molten waxes. On chamber of a fluidized bed unit. The capsules rotated
cooling, drug–wax plugs are obtained. Some of the during fluidization at temperatures above the melting
advantages of liquid filling of hard gelatin capsules, point of the wax and centrifugal forces caused the
when compared to solid-filled capsules, include a better drug–wax melt to flow into the ends of the capsules.
weight uniformity, and the elimination of dust hazards The molten mixture then solidified after ending the
Water–Zeta

and cross-contamination via airborne particles. The heating and two solid wax matrices with dissolved/
drug–carrier system should not interact with the gela- dispersed drug were obtained in the ends of the cap-
tin shell in the molten as well as in the congealed state, sules. Good control over the drug release was obtained
and the physical state of the drug and wax should not by using blends of waxes with different amphiphilic
change during storage. The melting point of the wax properties (HLB-values). Gelucire 50/13 (m.p. ¼
has to be low enough to avoid degradation of the drug 50
C, HLB ¼ 13) and Precirol ATO-5 (m.p. ¼ 53
C,


and damage to the capsule shell. C, HLB ¼ 2) were selected as the drug carriers.
Waxes 4071

The drug release increased with increasing proportion Coating with Hot Melts
of the more hydrophilic wax. After cooling of the
drug-containing wax melt, the drug could be dispersed, With regard to coating processes, various fluidized-bed
dispersed/dissolved, or dissolved in the wax matrix. techniques and their modifications were evaluated for
DSC studies were used to characterize the physical hot-melt coating.[38–40] The fluidized-bed techniques
state of the drugs after formation of the wax matrix. include the top, bottom, and the tangential spray or
A linear relationship existed between the heat of fusion rotary fluidized-bed modes. The particles to be coated
and the amount of drug in the wax matrix (Fig. 2). The are suspended in a heated high-velocity air stream, and
solubility of the drug in the matrix at its melting point the molten wax is applied in the form of atomized
corresponds to the intercept of the line. Propranolol liquid droplets.
HCl was insoluble and dispersed in the Precirol The top spray mode, in which the wax melt is
ATO-5 matrix while theophylline was partially dis- sprayed downward on upward moving particles, is
solved in the wax. the system of choice for hot-melt coating (Fig. 3).
The product temperature can be kept closest to the
congealing temperature of the wax when compared
Coating with Waxes
to the other two spray modes. The wax has to be kept
in a molten state in order to be atomized into the
Besides their predominant use in matrix preparations,
fluidized bed. A special nozzle had to be developed.
waxes have also been used as coatings for granules
The nozzle wand had a triaxial structure with a center
and pellets. Solid dosage forms are often coated to
tube for the molten liquid, which is surrounded by a
sustain the drug release, to improve the stability, or
small air space for the delivery of high-pressure, low-
to mask the taste of poorly tasting drugs. The coating
volume air to control the valve in the nozzle, which
with waxes has various advantages when compared to
opens when the pump was running. Both of these tubes
the coating with polymer solutions or dispersions.
were surrounded by a larger air space through which
The waxes can be applied without organic solvents,
the heated atomization air was supplied. The nozzle
and, in the case of hot melts, with a high application
should be placed as closely as possible to the substrate
rate and therefore shorter processing time. Many food
bed in order to minimize the distance that the molten
grade natural or semisynthetic waxy materials are
droplets had to travel prior to contacting the substrate
available.
surface.
Waxes can be applied onto solid dosage forms in the
Substrates with poor fluidization characteristics
form of hot melts, hot emulsions, aqueous suspensions
such as larger particles and/or particles of higher den-
(colloidal wax particles), or organic solutions. Coating
sity are difficult to coat with the top spray mode, and
processes include dip coating, pan coating, or fluidized-
the bottom spray mode should be preferred. With the
bed coating. The coating of drug particles by spray
tangential spray process, the product temperatures
congealing is discussed in the section on microencap-
have to be maintained lower than with the top spray
sulation with waxes.
technique in order to avoid adherence of the coated
particles to the product container. The tangential spray

50
y = 10.601 + 2.258X R = 0.99 Nozzle needle
Expansion Control air
chamber
40 Insulated Peristaltic Molten
nozzle pump coating
reservoir
30
% Drug

Thermometer
Heated
20 atomization air
y = 0.457 + 2.704X R = 1.00
Nozzle wand
10 Product cross section
Water–Zeta

container
Theophylline
Insulation
Propranolol HCl Heated atomization air
0
0 5 10 15 20 Control air
Cal/Drug-wax matrix (g) Molten coating liquid

Fig. 2 Relationship of propranolol HCl and theophylline Fig. 3 Insulated nozzle and wand for top-spray hot-melt
loading and the heat of fusion. (From Ref.[37].) coating. (From Ref.[38].)
 4072 Waxes

method is more stressful on the substrates than the melting point of less than 85
C because the melt is
other two techniques. usually kept at temperatures of 40–60
C above its
Important process and formulation variables melting point. Materials with a broad melting point
include the product bed temperature, the atomization range can become tacky during spraying because of
conditions, the type of substrate, the properties of the the broad range of product temperatures and the pres-
coating materials, and the desired release rates ence of low melting point fractions. The coating mate-
(immediate or sustained release). In order to obtain rials include various hydrogenated vegetable oils,
good coatings, the atomization air has to be heated beeswax, paraffin wax, carnauba wax, and polyethyl-
to the same temperature as the molten wax in order ene glycol.
to avoid premature congealing. The droplets must
remain in a liquid state until they hit the substrate sur- Coating with hot emulsions, aqueous
face. The product bed temperature is very critical to suspensions, or organic wax solutions
the successful coating of solid dosage forms with mol-
ten materials. At low product temperatures, premature The coating with hot emulsions has various advantages
congealing of the molten droplets results in poor when compared to the hot-melt process.[41,42] The wax
spreading of the coating on the substrate surface and, remains in the molten state and premature congealing
in the extreme case, in the failure of the adherence of of the wax could be eliminated because of the presence
the coating to the surface. Rough and porous surface of hot water. This facilitated the transport of the wax
structures are obtained, resulting in faster drug release to the spray nozzle and therefore the experimental
when compared to substrates with smooth coatings. At setup. In addition, the temperatures of the wax emulsion
too high product temperatures, excessive particle are lower then the comparable wax melts. o/w-
agglomeration or clogging of the outlet filter bags is Emulsions of various waxes (e.g., glyceryl behenate-
a result of inadequate congealing and hardening of Compritol-888 and glyceryl palmitostearate-Precirol-
the coating. The product bed temperature can be regu- ATO 5) with a solids content of up to 50% were
lated through the fluidization air temperature. It is prepared. The emulsions were passed through a micro-
recommended to use inlet air temperatures 10–15
C fluidizer to further reduce the particle size of the oil
below the melting point of the coating and tempera- phase. The hot emulsion was then either sprayed
tures for the atomization air and the molten wax of directly on the beads or cooled to form a ‘‘wax pseudo-
40–60
C above the melting point. latex’’ prior to the coating process. The disadvantages,
Droplet size and uniformity are also critical for a when compared to hot-melt coating, include the
successful coating process. The size of the molten drop- amount of water to be evaporated and therefore longer
let is dependent on the viscosity of the melt and the processing times, and the presence of surfactants in the
atomization air pressure. Smaller particles require wax coating. The surfactants, which were needed to
smaller droplet sizes and therefore higher atomization stabilize the emulsions, could affect the drug release.
air pressures in order to minimize agglomeration or The use of liquid surfactants such as various Tween/
granulation. In order to obtain small droplets, the vis- Span combinations resulted in sticky beads. Solid
cosity of the molten material can be decreased by surfactants such as sodium lauryl sulfate were more
increasing the temperature of the melt. At the same suitable. The guaifenesin release from coated pellets
atomization conditions, low feed rates also result in decreased with increasing hydrophobicity of the wax
smaller droplets. The spray rate of melts is generally and increasing coating level. The coating conditions
much lower when compared to coating solutions or (e.g., temperature, spray rate, curing) and the particle
dispersions. The lower spray rate, however, is offset size of the emulsion/suspension primarily influenced
by the application of pure coating material. The appli- the microstructure of the wax coatings and hence the
cation rate therefore is still higher when compared to drug release. When compared to polymer coatings,
polymer solutions or dispersions, with which solvents thicker coatings have to be applied with waxes to
have to be evaporated. obtain the same sustained release profiles.
After the application of the melt, the fluidization is Bagaria prepared emulsions with waxes including
reduced and the product bed is cooled. The cooling cir- carnauba, paraffin, ceresin, beeswax, and hydroge-
cle should be short in order to avoid attrition of the nated castor, soybean, or cottonseed oil, which were
Water–Zeta

coated product. Rapid cooling, however, may result then coated onto drug-containing non-pareil beads.[43]
in cracks in the coating because of contraction of the The term emulsions was actually misleading because
coating material, and it may also result in unstable the final products were aqueous suspensions of the
polymorphic forms of the wax. waxes with partially submicron particle size (wax
The important variables for the selection of the pseudolatex). Similar to aqueous polymer dispersions,
waxes include the melting point, the melting range, the wax dispersions were converted into powders by
and the viscosity of the melt. The wax should have a spray drying. The release from beads coated with the
Waxes 4073

redispersed spray dried powder was then compared to During emulsification, the drug will partition into the
beads coated with the original dispersion. With almost external aqueous phase until its solubility at the emul-
all preparations, 100% drug was released within 2–4 h. sification temperature is reached. During cooling of the
Like with organic polymer solutions, the wax can be emulsion, the drug could precipitate in the aqueous
dissolved in an organic solvent and then be sprayed phase because of a decreased drug solubility. The type
onto the solid dosage form. In most studies, waxes such of wax, the rate of cooling, the stirring time, and the
as beeswax, hydrogenated castor oil, microcrystalline temperature of the aqueous phase had no significant
wax, or glyceryl mono- and distearate were dissolved effect on the drug loading because of the low solubility
in chlorinated organic solvents such as chloroform, of ibuprofen in the external aqueous phase. Actual
carbon tetrachloride, or trichlorethane and applied in drug loadings close to 60% could be achieved. The
coating pans at elevated temperatures.[44–46] Mixtures drug release was controlled by the hydrophobicity of
of ethyl cellulose with different waxes such as castor, the wax (Gelucire 64/02 > Precirol ATO5 >
carnauba, or paraffin wax in chloroform were evalu- beeswax > carnauba wax > paraffin wax) (Fig. 4).
ated as sustained-release coatings.[47] A lower ratio of These wax microparticles could be formulated into
ethyl cellulose to wax was used with drugs of high- aqueous sustained-release oral suspension dosage
molecular weight and/or low solubility and a higher forms because of the low solubility of the drug.
ratio for drugs of low-molecular weight and/or A modified USP method using mini baskets was
high solubility to achieve the desired release properties used to study the effect of formulation variables, such
at a 10% coating level. As the date of the references as type of wax, type of modifier, drug loading, size, on
shows, the coating with organic solutions is obsolete the ibuprofen release from wax microspheres.[49] The
because of the undesirable use of organic solvents. drug release was in the order of beeswax > ceresine
wax > refined paraffin wax > microcrystalline wax.
After an initial burst release, the drug release was slow
Microencapsulation with Waxes from microparticles prepared with the last two waxes.
The dissolution studies were performed in simulated
Wax microparticles have been prepared primarily by intestinal fluids, because sink conditions could not be
aqueous and non-aqueous melt dispersion techniques maintained in simulated gastric fluids because of the
or spray congealing/spray drying. These techniques drug solubility. To increase the drug release from the
are briefly reviewed below. wax microspheres, glycerol monostearate or stearyl
alcohol were added prior to the preparation of the
Microparticles prepared by melt-dispersion microparticles. The addition of these release modifiers
techniques also reduced the tendency of the waxes to agglomer-
ate.[50] Ibuprofen dissolved in the molten wax and
In the melt dispersion technique, the drug-containing did not crystallize during congealing and microsphere
molten wax phase is emulsified into a heated, emulsifier- formation, as indicated by DSC studies.
containing external phase. Depending on the solubility
of the drug, the external phase can be either aqueous
(for water-insoluble drugs) or non-aqueous (for water- 100
soluble drugs). On cooling the emulsion, the liquid dro-
plets congeal and a suspension of the wax microparticles
75
Drug released (%)

is formed. The microparticles are then separated, mostly

by filtration or centrifugation, sometimes washed to
remove free drug crystals and surfactants, dried and
sized. 50
Ibuprofen-wax (carnauba wax, paraffin wax, bees-
wax, Gelucire 64/02, or Precirol ATO5) microparticles
were characterized with respect to drug loading and 25
morphological and release properties.[48] Microparti-
cles of the more hydrophilic waxes, Gelucire 64/02
Water–Zeta

and Precirol ATO5, could be prepared without surfac- 0

tants, while the other waxes rapidly coalesced and 0 6 12 18 24
formed big lumps on cooling. With the other waxes, Time (h)
increasing the amount of sodium lauryl sulfate in the Fig. 4 Ibuprofen release from different wax microparticles
external aqueous phase decreased the drug loading (actual drug loading): (&) Precirol ATO5 (35.3%), (&)
because of drug solubilization. This was not the Gelucire 64/02 (35.4%), (G) beeswax (37.7%), (`) carnauba
case when poly(vinyl alcohol) was used as stabilizer. wax (37.3%), (}) paraffin wax (35.0%). (From Ref.[48].)
 4074 Waxes

In a modified melt-dispersion method, sulfamethox- congealing and to assure the formation of an emulsion.
azole particles were dispersed in heated water, and The microparticles formed after congealing of the wax.
powdered beeswax was added. The molten wax drop- A key for high encapsulation efficiencies was the for-
lets collected the drug particles, and spherical agglom- mation of ultrafine internal aqueous phase droplets
erates with sustained-release properties were obtained by sonication. The wax acted as a diffusion barrier
after cooling. Alternatively, the agglomerates could also between the internal and the external aqueous phases
be formed at room temperature by using solutions of the and therefore minimized drug partitioning into the
wax in a water-immiscible solvent.[51] external aqueous phase. Because of the high solubility
Water-soluble drugs cannot be encapsulated with of the drug in the external aqueous phase, the contact
the O/W-emulsion technique because the drug would time of the droplets/microparticles with the continu-
be lost to the external aqueous phase. Two methods ous aqueous phase had to be minimized in order to
have been described for the encapsulation of hydrophi- avoid the drug loss; the microparticles were separated
lic drugs; one is based on using an external oil phase from the aqueous phase within minutes after their for-
and the other one on the formation of the microparti- mation.
cles by a w/o/w-melt dispersion technique. Generally, the wax phase is emulsified into a heated
Carnauba-wax microspheres containing 5-fluoro- external phase to avoid premature congealing. How-
uracil for chemoembolization were prepared by a melt ever, sulfamethazine-japanese synthetic wax particles
dispersion process with an external silicone oil were prepared by dispersing the drug in the molten
phase.[52] The drug was dispersed in the molten car- wax, followed by slowly pouring the wax phase into
nauba wax and emulsified into silicone oil at tempera- a precooled external aqueous phase.[54] The micropar-
tures above the melting point of carnauba wax (85
C). ticles were then separated by filtration after 3 min.
The resulting emulsion was cooled through the Beeswax microparticles were prepared by a phase
addition of cold silicone oil and immersion of the bea- inversion technique, whereby an aqueous solution of
ker in an ice-water bath. After solidification, the micro- sorbitan monooleate and polysorbate 80 was added
spheres were separated from the oil phase by to the drug-containing molten wax phase to first form
centrifugation and washed with cyclohexane to remove a w/o-emulsion prior to phase inversion. Smaller
the silicone oil. 5-Fluorouracil is a hydrophilic drug; an microparticles were obtained at higher temperatures,
external aqueous phase would have resulted in drug with increased amounts of continuous phase, with
partitioning and therefore low encapsulation efficien- slower rates of cooling, and with higher speeds of
cies. However, even with silicone oil, an external phase mixing.[55]
in which the drug was insoluble, only up to 5% drug Suspensions of lipid nanoparticles can be prepared
could be encapsulated within the carnauba-wax micro- by reducing the size of the molten or dissolved drug
spheres. This was attributed to the poor wetting of the containing lipid phase into the colloidal size range by
drug crystals by the molten wax and therefore the loss high-pressure homogenization.[56,57] After cooling or
of drug crystals to the external silicon oil phase. solvent evaporation, the nanoparticles are obtained.
Various surfactants were added to the wax phase in The suspensions of the nanoparticles can be converted
order to improve the wettability of the drug by the into powders through freeze or spray drying. The
molten wax. nanoparticles can act as carriers for poorly water-
A technique based on the formation of a multiple soluble drugs and apparently could result in controlled
emulsion with an external aqueous phase was release over longer periods of time. However, because
developed for the encapsulation of water-soluble drugs of the small size and therefore high surface area of
in order to replace the external oil phase.[53] Possible the particles, an increase in dissolution rate would be
unwanted interactions between the oil and the emulsi- expected unless very low loadings are used.
fied wax such as swelling or dissolution of the wax,
clean-up requirements of the final product, and recov- Microparticles prepared by spray drying
ery of the oil phase could be eliminated. In analogy to and spray congealing
the encapsulation of water-soluble drugs within poly-
meric microparticles by a w/o/w-solvent evaporation Similar to organic polymer solutions, drug-containing
method, a molten wax phase was used instead of an organic wax solutions were spray dried to give
Water–Zeta

organic polmer solution. A heated aqueous solution sustained-release microparticles.[58] The drug could be
of pseudoephedrine HCl was emulsified into the mol- either dissolved or dispersed in the organic wax solu-
ten carnauba wax, followed by the emulsification of tions. Spray drying is a single step, rapid drying
this w/o-emulsion into a heated external aqueous process, which can be scaled up and be used for
phase. The temperature of the internal and external heat-sensitive drugs. The use of organic solvents, how-
aqueous phases had to be kept above the melting ever, is undesirable because of solvent hazards, solvent
temperature of the wax in order to avoid premature residuals, and cost. Because of the low melt viscosity,
Waxes 4075

wax microparticles can also be prepared without melt, the lipid can crystallize in different polymorphic
organic solvents by spray-congealing drug-containing forms, depending on the composition of the lipid and
wax melts. thecooling rate. The major polymorphic forms of the
Sulfaethylthiadiazole-hydrogenated castor oil parti- glycerides are the a-, b-, and b0 -forms. Rapid cooling
cles were prepared by spray congealing, using a cen- ratesgenerally result in the unstable a-form. The tran-
trifugal wheel atomizer.[59] The wax powder was then sition of the melt first to the a- and b0 -forms and then
suspended into water to give an oral sustained-release to the b-form represents the transformation of trigly-
suspension. In a subsequent article, the effect of vari- cerides into the most stable form.
ous process and formulation variables on the particle The spraying process, especially the spray con-
size was investigated using a centrifugal wheel atom- gealing was simulated by cooling the molten samples
izer.[60] The particle size was directly proportional to rapidly at 320
C/min. A DSC thermogram of pure
the feed rate and inversely proportional to the feed tristearin heated at 10
C/min revealed a single
viscosity and the wheel velocity. endothermic peak representing the b-form. After rapid
In a series of publications, sulfaethylthiadiazole-wax cooling of the melt and reheating, an endothermic peak
microparticles prepared by spray-congealing were eval- for the a-form and anexothermic peak indicating the
uated. The most important factor affecting the drug recrystallization of the a-form into the b-form,
release was the type of wax used.[61] The effect of sur- followed by the endothermic peak for the b-form, were
factant on the drug release from spray congealed wax detectable. The DSC thermogram of spray-dried tri-
microparticles was investigated by John and Becker.[62] stearin micropellets was similar to the one of the
White wax-USP, a synthetic wax-like ester, and 1 : 1 melt-quenched sample. A crystalline modification
combinations of these two waxes were used as sustained- therefore took place during spray drying because of
release matrices. The particle size of the microparticles the rapid solvent removal. The micropellets were then
could be controlled through the nozzle size, with the stored at different temperatures to investigate the effect
larger nozzles resulting in larger particles with slower of storage temperature on the polymorphic transitions.
rates of dissolution. Up to 4% sorbitan monooleate Increasing the storage temperature to 37
C resulted in
apparently softened the particles and promoted wetting, a complete transformation of the unstable polymor-
thus resulting in an increase in drug release. However, at phic form into the stable polymorphic form. The melt-
a surfactant level of 10%, the slowest release rate was ing endotherm of the a-form disappeared. The effect of
observed. This was partly attributed to the tackiness of various emulsifiers such as lecithin or monoglycerides
the particles, which resulted in agglomeration and a was investigated in order to prevent or delay the trans-
reduction in total surface area available for drug release. formation of the unstable form into the stable b-form.
The addition of the surfactant allowed the compression Adding lecithin to the formulation resulted in a delay
of the microparticles into tablets, which was difficult of the transformation. The type of glyceride (compo-
with the surfactant-free microparticles.[63] The surfac- sition and chain length), solvent, and drugs encapsu-
tant-free microparticles adhered or stuck to the punch lated affected the polymorphic transformation, its
surface, and resulted in a high friability of the finished rate of transformation, and also the surface structure
tablets. No additives were incorporated into the tablets; of the microparticles. Spray-congealed lipid micropel-
non-disintegrating wax matrix tablets were obtained. lets showed a similar thermal behavior as the spray-
A palatable suspension of the bitter-tasting drug, dried pellets. The smooth surface of the sprayed lipid
remoxipride, was developed based on microparticles micropellets was attributed to the unstable polymor-
prepared by spray congealing.[64] Because of the high phic form. The unstable a-form has various thin and
water solubility of the drug, the microparticles were small crystals, resulting in a smooth surface of the
formulated into an external oil phase. Unfortunately, crystallized sample. The b- and b0 s-forms have larger
neither the nature of the wax nor of the oily vehicle crystals, therefore causing irregular structures of the
was revealed. micropellets. During ageing at elevated temperatures,
With lipid drug delivery systems, polymorphic the lipid micropellets lost their smooth surface struc-
transformations may occur during the preparation of tures because of polymorphic transformations.
the dosage form and during subsequent storage. The Novel oral controlled release microspheres using
polymorphic behavior of lipid micropellets prepared polyglycerolesters of fatty acids and hydrogenated
Water–Zeta

from glycerides and phospholipids by spray drying or cottonseed oil (HCSO), stearic acid, stearyl alcohol,
spray congealing and their surface structure were eval- or glycerol monostearate as carriers were prepared by
uated by DSC and scanning electron microscopy.[65–67] spray chilling using a rotating disc.[68] The drug release
The rapid solvent evaporation during spray drying can was related to the hydrophobicity of the wax; the
influence the crystallization of the lipid carrier and dif- release rate decreased in the following order: stearyl
ferent polymorphic structures could be obtained. Simi- alcohol > stearic acid > glycerol monostearate >
larily, during spray congealing and solidification of the carnauba wax > hydrogenated cottonseed oil. Adding
 4076 Waxes

increasing amounts of lactose to HCSO increased the 21. Huang, H.-P.; Mehta, S.C.; Radebaugh, G.W.; Fawzi, M.B.
drug release as a result of the leaching of lactose. Mechanism of drug release from an acrylic polymer-wax
matrix tablet. J. Pharm. Sci. 1994, 83, 795–797.
A new atomizer operating with ultrasonic energy 22. De Brabander, C.; Vervaet, C.; Fiermans, L.; Remon, J.P.
was used as an alternative to traditional atomizers to Matrix mini-tablets based on starch/microcrystalline wax
prepare wax microparticles by spray congealing.[69] mixtures. Int. J. Pharm. 2000, 199, 195–203.
23. Dakkuri, A.; Schraeder, H.G.; DeLuca, P.P. Sustained
release from inert wax matrixes. II: effect of surfactants
on tripelennamine hydrochloride release. J. Pharm. Sci.
1978, 67, 354–357.
24. Foster, T.P.; Parrott, E.L. Constant release rate from inert
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Water–Zeta
 Wet Granulation: End-Point Determination
and Scale-Up
Michael Levin
Metropolitan Computing Corporation, East Hanover, New Jersey, U.S.A.

INTRODUCTION control the granule size and granulation rate. The end-
point controls the mix consistency and reproducibility.
Wet granulation is used mainly to improve flow and Other factors that affect the granule quality include
compressibility of powders, and to prevent segregation spray position and spray nozzle type, and of course,
of the blend components. Particle size of the granulate the product composition. Such variables as mixing
is affected by the quantity and feeding rate of the time and bowl or product temperature are not inde-
granulating liquid. pendent factors in the process but rather are responses
Wet massing in a high shear mixing is frequently of the primary factors listed above.
compared to the fluid bed mixing and to the roller
compaction technique,[1] and the results seem to be for-
mulation dependent. Compared to high shear granu- WHAT IS AN END-POINT?
lation, low shear or fluid bed process requires less
fluid binder, not only resulting in a shorter drying time, End-point can be defined by the formulator as a target
but also in a less cohesive material.[2–4] particle size mean or distribution. Alternatively, the
For excellent classical review of the wet granulation end-point can be defined in rheological terms. It has
process, equipment and variables, and measurement been shown[15] that once you have reached the desired
instruments available in the field, see papers by Holm end-point, the granule properties and the subsequent
et al.[5–12] These papers have become a standard refer- tablet properties are very similar regardless of the
ence for numerous subsequent publications. granulation processing factors, such as impeller or
Because of rapid densification and agglomeration that chopper speed or binder addition rate. I would call this
are caused by the shearing and compressing action of the ‘‘the principle of equifinality.’’
impeller in a high shear single pot system, mixing, granu- The ultimate goal of any measurement in a granu-
lation, and wet massing can be done relatively quickly lation process is to estimate the viscosity and density
and efficiently. The dangers lie in the possibility of over- of the granules, and, perhaps, to obtain an indication
granulation because of excessive wetting and producing of the particle size mean and distribution. One of the
low porosity granules, thus affecting the mechanical ways to obtain this information is by measuring the
properties of the tablets. As the liquid bridges between load on the main impeller.
the particles are formed, granules are subjected to Mixer instrumentation, in general, has numerous
coalescence alongside with some breakage of the bonds. benefits. In addition to a possible end-point determi-
It stands to reason that mean granule size is strongly nation, it can be used to troubleshoot the machine per-
dependent on the specific surface area of the excipients, formance (for example, help detect worn-out gears and
as well as the moisture content and liquid saturation of pulleys or identify mixing and binder irregularities).
the agglomerate. During the wet massing stage, gran- Instrumentation can serve as a tool for formulation
ules may increase in size to a certain degree while the fingerprinting, assure batch reproducibility, aid in raw
intragranular porosity goes down. However, some material evaluation, process optimization, and scale-up.
heating and evaporation may also take place leading
to a subsequent decrease in the mean granule size,
especially in small-scale mixers. WHAT CAN BE MEASURED ON A
Water–Zeta

Load on the main impeller is indicative of granule MIXER-GRANULATOR?

apparent viscosity and wet mass consistency. It can
be seen as an interplay of acceleration (direct impact Current
of the impeller), centrifugal, centripetal, and friction
forces that act on the particles. Current in direct current (DC) motors can be used as
According to Cliff,[13,14] binder addition rate con- some indication of the load on the main impeller
trols granule density, while impeller and chopper speed because impeller torque is proportional to current in
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-120014089
4078 Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved.
Wet Granulation: End-Point Determination and Scale-Up 4079

some intervals[13] and therefore, a current meter Impeller or Motor Shaft Speed
(ammeter) can be used for small scale DC motors.
However, for alternating current motors (most often Rate of impeller rotation could be used as some indi-
used in modern mixers), there may be no significant cation of the work being done on the material.[24] As
change in the current as motor load varies up to 50% the motor or impeller power consumption is pro-
of full scale. At larger loads, the current draw may portional to the product of torque and speed, the latter
increase but this increase is not linearly related to load, is an important factor in evaluating the corresponding
and, consequently, the current is completely ineffective load.
as a measurement of load. Moreover, the current base-
line may shift with time.
Motor Slip and Motor Load Analyzer

Voltage Motor slip is the difference between the rotational

speed of an idle motor and the motor under load.[25,26]
Voltage measurement generally has no relation to load. Motor slip measurements, although relatively inex-
pensive, do not offer advantages over the power
Capacitance consumption measurements. The method did not
gain popularity, probably because the slip is not
Capacitive sensor responds to the moisture distri- linearly related to the load,[27] despite some claims to
bution and granule formation.[16–19] It provided similar the contrary.
end-points (based on the total voltage change) under
varying rates of agitation and liquid addition. Capaci-
Impeller Tip Speed
tive sensor can be threaded into an existing thermo-
couple port for in-process monitoring.
Impeller tip speed corresponds to the shear rate and
has been used as a scale-up parameter in fluid mix-
Conductivity ing.[28] For processing of lactose granulations in Gral
mixers, however, it was shown by Horsthuis et al.[29]
Conductivity of the damp mass[20] makes it possible to that the same tip speed did not result in the same end-
quantify uniformity of liquid distribution and packing point [in terms of particle size distribution (PSD)].
density during wet massing time. These findings were contradicted by other studies with
Fielder mixers indicating that for a constant tip speed,
successful scale-up is possible when liquid volume is
Probe Vibration
proportional to the batch size and wet massing time
is related to the ratio of impeller speeds.[30]
Probe vibration analysis[21,22] requires a specially con-
structed probe that includes a target plate attached to
an accelerometer (for in-process monitoring). This Relative Swept Volume
measurement is based on the theory that increasing
granule size results in the increase of the acceleration Relative swept volume, that is, the volume swept by the
of agglomerates striking the probe target. The method impeller (and chopper) per unit time, divided by the
has a potential for granulation monitoring and end- mixer volume, has been suggested as a scale-up
point control. factor.[11,12,31] This parameter is related to work done
on the material and was studied extensively at various
Boots Diosna Probe blade angles.[32] Higher swept volume leads to higher
temperature and denser granules. However, it was
This probe[23] measured the densification and increase shown by Horsthuis et al.[29] that the same relative
in size of the granules (changes in momentum of gran- swept volume did not result in the same end-point (in
ules moving with constant velocity because of a mass terms of PSD).
change of the granules). The method did not gain
Water–Zeta

popularity because of its invasive nature.

Temperature

Chopper Speed Product and jacket temperature are usually measured

by thermocouples. These response variables are con-
Chopper speed has no significant effect on the mean trolled by a variety of factors, notably, the speed of
granule size.[5,6] the main impeller and the rate of the binder addition.
 4080 Wet Granulation: End-Point Determination and Scale-Up

Binder Addition Rate Motor power consumption is a product of current,

voltage, and the so-called power factor. In the range
There are conflicting reports on the preferred method of interest, motor power consumption is generally pro-
of adding the binder. For example, Holm[32] does not portional to load on the motor and, to some degree,
generally recommend adding dry binder to the mix can reflect the load on the impeller (Fig. 1).
(as commonly done in order to avoid preparation of However, up to 30% of the power consumption of a
a binder solution) because homogeneity of binder dis- motor can be attributed to no-load losses because of
tribution cannot be assured. Others recommend just windage (by cooling fan and air drag), friction in the
the opposite.[34–36] bearings, and core losses that comprise hysteresis and
Slow continuous addition of water (in case the water- eddy current losses in the motor magnetic circuit. Load
soluble binder is dry mixed) or a binder solution to the losses include stator and rotor losses (resistance of
mix is a granulation method of choice.[5,6,10–12,37–44] materials used in the stator, rotor bars, magnetic steel
The granulating fluid should be added at a slow rate to circuit) and stray load losses such as current losses in
avoid local overwetting.[33] the windings.[58]
If the binder solution is added continuously, then Attempts to use a no-load (empty bowl, or dry mix)
the method of addition (pneumatic or binary nozzle, value as a baseline may be confounded by a possible
atomization by pressure nozzle) should be considered nonlinearity of friction losses with respect to the
in any end-point determination and scale-up. load.[59] As the load increases, so does the current draw
An alternative to a continuous binder liquid addition of the motor. This results in heat generation that
method is to add binder liquid all at once[29] to assure the further impacts the power consumption.[27] A simple
ease of processing and reproducibility, reduce processing test might be to run an empty mixer for several hours
time, and to avoid wet mass densification that may occur and see if there is any shift in the baseline. Also, as
during the liquid addition. This latter phenomenon the motor efficiency drops with age, the baseline most
may obscure the scale-up effect of any parameter under definitely shifts over time.
investigation. Motor power consumption is non-linearly related to
the power transmitted to the shaft[60] and the degree of
this non-linearity could only be ‘‘guestimated.’’
Power Consumption

One of the most popular and relatively inexpensive Impeller Torque

measurements is the power consumption of the main
mixer motor. It is measured by a watt transducer or In a mixing process, changes in torque on the blades and
a power cell utilizing Hall effect (a measurable trans- power consumption of the impeller occur as a result of
versive voltage between the two radial sides of a cur-
rent conductor in a magnetic field, an effect
discovered by E.H. Hall in 1879).
Power consumption of the mixer motor for end-
point determination and scale-up is widely used
(Leuenberger[43] and subsequent work, Holm[5] and
subsequent work, Landin et al.[45–48], Faure et al.[49–53],
and many others[16,33,37–44,54,55]) because the measurement
is economical, does not require extensive mixer modifica-
tions, and is well correlated with the granule growth.
Power consumption correlates with the mean gran-
ule size of a granulation,[8] although the correlation is
Power
not always linear in the entire range. Intragranular
porosity also shows some correlation with power con- Current

sumption.[56] Normalized work of granulation (power Voltage

profile integrated over time) can accurately determine

Water–Zeta

end-points and is correlated well with properties of

the granulates.[57]
The main problem with the power consumption
measurements is that this variable reflects load on the ldle Full
Load
motor rather than load on the impeller. It relates to the
overall mixer performance, depends on the motor Fig. 1 Voltage, current, and power consumption of a typical
efficiency, and can change with time regardless of the load. mixer motor.
Wet Granulation: End-Point Determination and Scale-Up 4081

change in the cohesive force or the tensile strength of the viscosity,[67] it can have a conditional use in the dimen-
agglomerates in the moistened powder bed. sional analysis of the process, as will be shown below.
Direct torque measurement requires installation of
strain gages on the impeller shaft or on the coupling
between the motor and impeller shaft (Fig. 2). As the Reaction Torque
shaft is rotating, a device called slip ring is used to
transmit the signal to the stationary data acquisition By the third law of Newton, for every force there is a
system. counterforce, collinear, equal and opposite in direc-
Planetary mixer instrumentation for direct torque tion. As the impeller shaft rotates, the motor tries to
measurement does not substantially differ from that rotate in the opposite direction, but it does not because
of a high shear mixer. Engineering design should only it is bolted in place. The tensions in the stationary
take into account the planetary motion in addition to motor base can be measured by a reaction torque
shaft rotation.[61] transducer.
Impeller torque is an excellent in-line PAT measure- Reaction torque is a less expensive alternative to
ment of the load on the main impeller.[39,62] direct impeller torque and is recommended for mixers
that have the motor and impeller shafts axially aligned
(in this case, the reaction torque is equal to direct tor-
que and is opposite in sign).
Torque Rheometer

A torque rheometer is a device that provides an off-line

Other Possibilities
measurement of torque required to rotate the blades of
the device and this torque can be used to assess rheolo-
When the agglomeration process is progressing very
gical properties of the granulation. It has been exten-
rapidly, neither power consumption nor torque on
sively used for end-point determination.[45,63–65] The
the impeller may be sensitive enough to adequately
torque values thus obtained were termed as ‘‘measure
reflect changes in the material. Some investigators feel
of wet mass consistency.’’[50,51,66]
that other measurements, such as torque or force on
One of the main concerns is that using the torque
the impeller blades may be better suited to monitor
value that the unit is reporting instead of the dynamic
such events.
viscosity for calculation of Reynolds numbers renders
There are other ideas floating around, for example,
the latter to become dimensional. Therefore, the
use of neural network to describe and predict the beha-
Reynolds number calculated from torque rheometer
vior of the wet granulation[68] or control the end-point
data is referred to as ‘‘pseudo-Reynolds’’ dimensional
by rapid image processing system.[69]
number. Because of the fact that torque was shown to
A technique for measuring tensile strength of gran-
be proportional to a kinematic (rather than dynamic)
ules, in addition to power consumption measurement,
to facilitate optimal end-point determination was recently
described by Betz, Bürgin, and Leuenberger.[54]
Powder flow patterns in wet granulation can be
studied using positron emission particle tracking.[70]
Eventually, this and similar techniques can be used to
validate various mathematical and statistical models
of the process.

Emerging Technology

Acoustic

Applicability of piezoelectric acoustic emission sensors

Water–Zeta

to end-point determination has been studied since the

beginning of this century.[71] The technique is very
promising, especially because it is non-invasive, sensi-
tive, and relatively inexpensive. Granulation process
signatures obtained with acoustic transducer can be
used to monitor changes in particle size, flow, and
Fig. 2 Schematic of a direct impeller torque transducer. compression properties.[72,73]
 4082 Wet Granulation: End-Point Determination and Scale-Up

Near-infrared END-POINT DETERMINATION

Use of a refractive near-infrared (NIR) moisture sen- End-point detection in wet granulation has become a
sor for end-point determination of wet granulation major scientific and technological challenge.[78] Moni-
was described by several authors.[74,75] There are tech- toring granulation is most commonly achieved by col-
nological challenges associated with this approach, as lecting either power or torque signals, or both. In what
the sensor can only measure the amount of water at follows, we will compare both methods.
the powder surface.
NIR monitoring of the granulation process was
attempted by researchers at many major pharmaceuti- Torque vs. Power
cal corporations with a modest success. In particular,
yet unpublished work by David Rudd of Glaxo- When we say, ‘‘power consumption,’’ we usually refer
SmithKline in England should be mentioned as a part to the main motor. It reflects the load on the motor
of the global effort in the field of Process Analytical because of useful work, as well as the power needed
Technology. to run the motor itself (losses because of eddy currents,
friction in couplings, etc.).
It is quite possible (and, indeed, quite pertinent) to
Focused beam reflectance talk about the power consumption of the impeller,
measurement which is, obviously, quantitatively less than the power
consumption of the motor and relates directly to the
Focused beam reflectance measurement (FBRM) is a load on the impeller.
particle size determination technique based on a laser
beam focusing in the vicinity of a sapphire window Power
Torque  Speed
of a probe. The beam follows a circular path at speeds
of up to 6 m/sec. When it intersects with the edge of a Impeller power consumption can be calculated as a
particle passing by a window surface, an optical collec- product of the direct torque, rotational impeller speed,
tor records a backscatter signal. The time interval of and a coefficient (usually equal to 2p times a unit
the signal multiplied by the beam speed represents a conversion factor, if required).
chord length between two points on the edge of a par- The power consumption of the mixer motor differs
ticle. The chord length distribution (CLD) can be from that of the impeller by the variable amount of
recalculated to represent either a number or volume power draw imposed by various sources (mixer
weighted PSD. condition, transmission, gears, couplings, motor con-
In many cases, where precision is more important dition, etc.).
than accuracy, CLD measurements are adequate to Compared to impeller torque, motor power con-
monitor dynamic changes in process parameters sumption is easier to measure; wattmeters are inexpen-
related to the particle size and shape, concentration, sive and can be installed with almost no downtime.
and rheology of fluid suspensions. However, motor power signal may not be sensitive
Several attempts were made to evaluate the use enough for specific products or processing conditions.
of FBRM particle size analyzer as a potential tool Wear and tear of mixer and motor may cause power
for granulation end-point determination.[76] Dilworth fluctuations. Moreover, power baseline may shift with
et al.[77] have compared power consumption, FBRM, load.
and acoustic signals in a study of a wet granulation Impeller torque, on the other hand, is closer to
process in Fielder PMA 200 mixer. It was found that where the action is, and is directly related to the
these techniques were complimentary, with FBRM load on the impeller. Torque is not affected by mixer
probe capable to follow median granule size growth condition.
even when the power consumption curve showed a Although the motor power consumption is strongly
plateau. correlated with the torque on the impeller,[39] it is less
A major disadvantage of the FBRM method is that sensitive to high frequency oscillations caused by direct
the measured CLD does not directly represent a PSD. impact of particles on the blades as evidenced by Fast
Water–Zeta

Conversion of CLD to PSD is not straightforward and Fourier Transform (FFT) technique.[16]
requires sophisticated mathematical software that is Power consumption or torque fluctuations are influ-
not easy to validate. Moreover, CLD depends on enced by granule properties (PSD, shape index, and
optical properties and shape of the particles, as well apparent density) and the granulation time. Fluctua-
as the focal point position. The total number of counts tion of torque/power consumption and intensity of
measured is a function both of solids concentration spectrum obtained by FFT analysis can be used for
and probe location. end-point determination.[38]
Wet Granulation: End-Point Determination and Scale-Up 4083

It was observed that when the end-point region of The area under the torque-time curve is related to
a granulation is reached, the frequency distribution of the energy of mixing and can be used as an end-point
a power consumption signal reaches a steady state.[79] parameter. Area under power consumption curve
It should be repeated here that torque shows more divided by the load gives the specific energy consumed
sensitivity to high frequency oscillations. by the granulation process. This quantity is well corre-
lated with the relative swept volume.[11,12,32]
The consumed energy is completely converted into
Torque and Power Profiles heat of the wet mass,[7] so that the temperature rise
during mixing shows some correlation with relative
Fig. 3 illustrates the classical power and torque swept volume and Froude number[29] that relates the
profiles that start with a dry mixing stage, rise steeply inertial stress to the gravitational force per unit area
with binder solution addition, level off into a plateau, acting on the material.
and then exhibit overgranulation stage. The power Fig. 4 represents a record of a typical granulation
and torque signals have similar shape and are strongly batch done by an experienced operator on large
correlated. The pattern shows a plateau region where Hobart mixer. You can see that the batch was stopped
power consumption or torque is relatively stable. on the downslope of the derivative.
The peak of the derivative indicates the inflection On a Fig. 5 you can see another batch made by the
point of the signal. Based on the theory by Leuenberger same operator. This time it is a power consumption
(1979 and subsequent work), usable granulates can be trace, but again it extends beyond the peak of the
obtained in the region that starts from the peak of the derivative and the end-point thus can be deemed repro-
signal derivative with respect to time and extends well ducible.
into the plateau area.[44] Prior to the inflection point, In the batch represented in Fig. 6, a novice operator
a continuous binder solution addition may require trainee has stopped the batch well before the peak of
variable quantities of liquid. After that point, the pro- the derivative. This required a major adjustment of
cess is well defined and the amount of binder solution the tableting operation (force and speed) to produce
required to reach a desired end-point may be more or tablets in an acceptable range of material properties
less constant. (hardness and friability).
Torque or power consumption pattern of a mixer In this batch (Fig. 7), the same novice operator has
is a function of the viscosity of both the granulate stopped the granulation process, opened the lid, took a
and binder. With the increasing viscosity, the plateau is sample, and decided to granulate for another 10 sec.
shortened and sometimes vanishes completely, thereby You can see that there is no indication that the peak
increasing the need to stop the mixer at the exact of the derivative was reached at the end-point.
end-point. Thus, it seems that the monitoring torque or power
At low impeller speeds or high liquid addition rates, can fingerprint not only the product, but the process
the classic S-shape of the power consumption curve and the operators as well.
may become distorted with a steep rise leading into A number of publications relate to the practical
overgranulation.[9] experience of operators on the production floor.[33,80–82]

200 3.5

Torque
Power 3
Torque slope
150 Power slope
2.5
Torque, N-m

Power, kW

Start of 2
binder addition
100
1.5
Water–Zeta

1
50

0.5

0 0 Fig. 3 Impeller torque and motor

0 50 100 150 200 power consumption for a small high
Time, sec shear mixer (Fielder PMA 10).
 4084 Wet Granulation: End-Point Determination and Scale-Up

Batch No. JJ0202A Batch No. JJ0214A

Torque
Torque slope Power
Power slope
800
6.0
Torque, lb-inch

Power, kW
600

4.0

400
Start of Start of
binder addition
binder addition
2.0
200

0 50 100 150 200 0 50 100 150 200 250 300

Time, sec
Time, sec
Fig. 6 A batch by a novice operator (power consumption
Fig. 4 A torque profile in a typical production batch.
profile).

End-Point Optimization end-point and the tableting processing parameters,

such as compression force or tablet press speed.
Agglomeration of particles in wet granulation have In one of the most interesting works based on the
been studied extensively.[24,83] The optimal end-point experimental design approach, an attempt was made
can be thought of as the factor affecting a number of to find a statistical relationship between the major
agglomerate properties (Fig. 8). factors affecting both granulation and compaction,
With so many variables involved in a granulation namely, granulation end-point, press speed (dwell time),
process, it is no wonder that more and more research- and compression force.[93] The resulting equation
ers throw in a number of factors together in an attempt allowed optimization of such standard response param-
to arrive at an optimum response.[34,84–92] eters as tablet hardness, friability, and disintegration
The final goal of any granulation process is a solid time. This study has also investigated the possibility of
dosage form, such as tablets. Therefore, when optimiz- adjusting the tableting parameters in order to account
ing a granulation process, the list of factors affecting for an inherent variability of a wet granulation process.
tablet properties may include both the granulation Multivariate optimization of wet granulation may
include hardness, disintegration, and ejection as
response variables.[94] Compressibility property of

Batch No. JJ0202C

Power
Power_slope
8.0
Power, kW

6.0

4.0
Start of
Water–Zeta

binder addition

2.0

0 50 100 150
Time, sec

Fig. 5 Another batch by the same operator (power consump- Fig. 7 Another batch by inexperienced operator (torque
tion profile). profile).
Wet Granulation: End-Point Determination and Scale-Up 4085

theoretical foundation) useful scale-up parameters of

the wet granulation process.[50,91,97]
In a seminal and elegant work published in 1993,
Horsthuis et al. from Organon in The Netherlands
have studied granulation process in Gral mixers of
10, 75, and 300 L size.[29] Comparing relative swept vol-
ume, blade tip speed and Froude numbers with respect
Mean Particle Size to end-point determination (as expressed by the time
Disintegration after which there is no detectable change in particle
Friability size), they have concluded that only constant Froude
Optimum Hardness numbers result in a comparable end-point.
End Point Level In another attempt to determine good scale-up
parameters, the University of Maryland group under
Fig. 8 Wet granulation end-point as a factor in tableting the direction of Dr. Larry Augsburger[30] has applied
optimization. the ideas of Leuenberger and Horsthuis to show that,
for a specific material, end-point can be expressed in
terms of wet massing time. For a constant ratio of a
granulations is extremely sensitive to various proces- binder volume to a batch size, this factor was found
sing parameters of wet granulation.[95] to be inversely proportional to impeller speed when
Recently, the experimental design procedure was the impeller tip speed was held constant for all batches.
applied to low shear wet granulation[96] with a factorial However, this result was not corroborated by other
design used to evaluate the influence of such factors as studies or other materials.
binder strength and agitator speed. Yet another example of semiempirical scale-up
effort[97] was based on the fact that normalized power
profiles are very similar and allow for direct compari-
End-Point Reproducibility son of different size granulators, at least for the equip-
ment and materials used in the study. Normalized
As will be shown in the following section, for every power curve rose at a relatively constant rate in the
blend and a fixed set of values for processing factors region where the ratio of water to dry mass is 0.1–0.2
(such as mixer geometry, blade speed, powder volume, (‘‘slope plateau’’). Despite a rapid increase in the slope
amount, and method of addition of granulating of the power curve, the desired end-point was still
liquid), a wet granulation process state (end-point) is detectable at a moment when the slope of the power
completely characterized by rheological properties of consumption signal exceeded the plateau level by a fac-
the wet mass (density, viscosity), which are, in turn, a tor of 5 (empirical observation). Using this approach,
function of particle size, shape, and other properties. an acceptable end-point (target particle size of
The process can be quantified with the help of dimen- 135 mm) was first established on a 10-L Fielder and
sionless Newton Power Number Np that will assume a then scaled to 65-L Fielder and 250-L Diosna.
certain numerical value for every state (condition) of
the granulate. Under fixed processing conditions,
Np will be proportional to Net Power Consumption Dimensional Analysis
DP for any end-point (defined, in part, by wet mass
density). Thus, in order to reproduce an end-point, it A rational approach to scale-up using dimensional
is sometimes sufficient to monitor power of the analysis has been in use in chemical engineering for
impeller (or the motor) and stop when a predefined quite some time. This approach, based on the use of
net level of the signal is reached. If, however, any of process similarities between different scales, was being
the processing variables or the rheological definition applied to pharmaceutical granulation since the early
of the end-point has changed, a more sophisticated work of Leuenberger in 1979.[43]
approach is required, as described below. Dimensional analysis is a method for producing
dimensionless numbers that completely describe the
Water–Zeta

process. The analysis should be carried out before the

END-POINT SCALE-UP measurements are made because dimensionless num-
bers essentially condense the frame in which the mea-
Scale-Up Attempts surements are performed and evaluated. The method
can be applied even when the equations governing
Numerous studies were undertaken in an attempt the process are not known. Dimensional analytical
to determine empirically (and, lately, with a solid procedure was first proposed by Rayleigh in 1915.[98]
 4086 Wet Granulation: End-Point Determination and Scale-Up

Principle of Similitude Newton (power) number, which relates the drag

force acting on a unit area of the impeller and the iner-
Imagine that you have successfully scaled up from a tial stress, represents a measure of power requirement
10-L batch to 300-L batch. What exactly happened? to overcome friction in fluid flow in a stirred reactor.
You may say: ‘‘I got lucky.’’ Apart from luck, there In mixer-granulation applications, this number can
had to be similarity in the processing and the end-point be calculated from the power consumption of the
conditions of the wet mass of the two batches. impeller or estimated from the power consumption of
According to the modeling theory, two processes the motor.
may be considered similar if there is a geometrical, Froude Number[101] has been described for powder
kinematic, and dynamic similarity.[99] blending and was suggested as a criterion for dynamic
Two systems are called geometrically similar if they similarity and a scale-up parameter in wet granulation.[29]
have the same ratio of characteristic linear dimensions. The mechanics of the phenomenon was described as
For example, two cylindrical mixing vessels are geome- interplay of the centrifugal force (pushing the parti-
trically similar if they have the same ratio of height to cles against the mixer wall) and the centripetal force
diameter. produced by the wall, creating a ‘‘compaction zone.’’
Two geometrically similar systems are called kine- Reynolds numbers relate the inertial force to the
matically similar if they have the same ratio of veloci- viscous force.[102] They are frequently used to describe
ties between the corresponding system points. Two mixing processes and viscous flow, especially in chemi-
kinematically similar systems are dynamically similar cal engineering.[103]
when they have the same ratio of forces between We have seen that there exists a sort of ‘‘principle of
the corresponding points. Dynamic similitude for wet equifinality’’ that states: ‘‘An end-point is an end-point
granulation would imply that the wet mass flow is and end-point, no matter how it was obtained.’’
patterns in the bowl are similar. Different processing pathways can lead to different
The gist of dimensionless analysis is as follows: For end-points, each with its own set of granulation pro-
any two dynamically similar systems, all the dimen- perties. However, once an end-point is reached, it is
sionless numbers necessary to describe the process characterized by certain numerical values of the
have the same numerical value.[100] Once a process is dimensionless variables describing the process, and
expressed in terms of dimensionless variables, we are these values will be independent of scale.
magically transferred to a world where there is no At the same end-point, no matter how defined, the
space and no time. Therefore, there is no scale and, rheological and dimensional properties of the granules
consequently, there are no scale-up problems. The pro- are similar. As we will see from the examples described
cess is characterized solely by numerical values of the below, that means that the density and dynamic vis-
dimensionless variables (numbers). In other words, cosity of the wet mass are constant, and the only vari-
dimensionless representation of the process is scale- ables that are left are the process variables, namely
invariant. batch mass, impeller diameter and speed, and the
Lack of geometrical similarity often is the main geometry of the vessel.
obstacle in applying the dimensional analysis to solv-
ing the scale-up problems. It was shown, for example,
that Collette Gral 10, 75, and 300 are not geometrically Comparison of Attainable Froude Numbers
similar.[29] In such cases, a proper correction to the
resulting equations is required. Horsthuis et al.[29] showed that an end-point could be
reproduced and scaled up in Gral mixers by keeping
the Froude numbers constant. For the same end-point,
Dimensionless Numbers in dynamically similar mixers (same geometrical ratios,
and same flow patterns), all dimensionless numbers
Dimensionless numbers most commonly used to describing the system should have the same numerical
describe the wet granulation process are Newton, value, but Froude numbers for any mixer are easiest to
Froude, and Reynolds: compute.
Each mixer has a range of attainable Froude num-
Water–Zeta

Np ¼ DP=ðr  n3  d5 Þ NewtonðpowerÞ bers, and an end-point transfer between mixers can only
be achieved when such ranges overlap. Fig. 9 repre-
Fr ¼ n2  d=g Froude
sents such a range for Collette Gral mixers. It can be
2
Re ¼ d  n  r=Z Reynolds seen that Gral 10 and Gral 150 have no overlap of
Froude number ranges, and therefore, a direct scale-
(for list of symbols, notation, and dimensions, see up is not possible (in addition, Gral mixers are not
Appendix). exactly similar geometrically, as was stated elsewhere).
Wet Granulation: End-Point Determination and Scale-Up 4087

2. Match these numbers at different scales.

The dimensionless space in which the measurements

are presented or measured will make the process ‘‘scale
invariant.’’

Relevance List

Fig. 9 The range of Froude numbers for Collette Gral high- The dimensional analysis starts with a list of all vari-
shear mixers. ables, thought to be important for the process, being
analyzed (the so-called ‘‘relevance list’’).
To set up a relevance list for any process, one needs
The range of Froude numbers for Fielder PMA to compile a complete set of all relevant and mutually
series mixers is shown on Fig. 10. The 10-L laboratory independent variables and constants that affect the
scale mixer at its lowest speed settings can reach the process. The word ‘‘complete’’ is crucial here. All
Froude numbers of all other mixers, except one. These entries in the list can be subdivided into geometric,
considerations can be useful for planning a scale-up or physical, and operational. Each relevance list should
technology transfer operation. include only one target (dependent ‘‘response’’) vari-
able.
Many pitfalls of dimensional analysis are associated
P-theorem (Buckingham)
with the selection of the reference list, target variable,
or measurement errors (e.g., when friction losses are
The so-called P-theorem (or Buckingham theorem)[104]
of the same order of magnitude as the power consump-
states:
tion of the motor). The larger the scale-up factor, the
Every physical relationship between n dimensional
more precise the measurements of the smaller scale
variables and constants f(x0, x1, x2, . . . , xn) ¼ 0 can
have to be.[100]
be reduced to a relationship f(P0, P1, . . . ,Pm) ¼ 0
between m ¼ n  r mutually independent dimension-
less groups, where r ¼ number of dimensional units,
i.e., fundamental units (rank of the dimensional matrix). Dimensional Matrix

Dimensional analysis can be simplified by arranging all

Scientific Scale-Up Procedure relevant variables from the relevance list in a matrix
form, with a subsequent transformation yielding the
1. Describe the process using a complete set of required dimensionless numbers. The dimensional
dimensionless numbers, and matrix consists of a square core matrix and a residual

Water–Zeta

Fig. 10 The range of Froude numbers for Fielder PMA high-shear mixers.
 4088 Wet Granulation: End-Point Determination and Scale-Up

Table 1 The relevance list used by Leuenberger (1983) One target variable (Power consumption) and seven
Quantity Symbol Units Dimensions process variables/constants thus represent the number
n ¼ 8 of the P-theorem. The number of basic dimen-
Power consumption P Watt M L2 T3
sions r ¼ 3 (M, L, and T). According to the theorem,
Specific density r kg/m3 M L3 the process can be reduced to the relationship between
Blade diameter d m L m ¼ n  r ¼ 8  3 ¼ 5 mutually independent
Blade velocity n rev/sec T1 dimensionless groups.
Binder amount s kg M The Dimensional Matrix in Table 2 was constructed
Bowl volume Vb m 3
L3 as described above, with the rows listing the basic
dimensions and the columns indicating the physical
Gravitational g m/sec 2
L T2
constant
quantities from the relevance list.
Transformation of the dimensional matrix (Table 3)
Bowl height H m L
into a unity matrix is straightforward. To transform-3
in L-row/r-column into zero, one linear transform-
ation is required. The subsequent multiplication of
the T-row by 1 transfers the 1 of the
matrix (you will see examples in the case studies n-column to þ1.
below). The five dimensionless groups are formed from the
The rows of the matrix consist of the basic dimen- five columns of the residual matrix by dividing each
sions, while the columns represent the physical quan- element of the residual matrix by the column headers
tities from the relevance list. The most important of the unity matrix, with the exponents indicated in
physical properties and process-related parameters, as the residual matrix.
well as the ‘‘target’’ variable (that is, the one we would The residual matrix contains five columns; therefore
like to predict on the basis of other variables) are five dimensionless P groups (numbers) will be formed
placed in one of the columns of the residual matrix. (Table 4).
The core matrix is then linearly transformed into a The end result of the dimensional analysis is an
matrix of unity where the main diagonal consists only expression of the form
of ones and the remaining elements are all zero. The
dimensionless numbers are then created as a ratio of
the residual matrix and the core matrix with the expo- P0 ¼ fðP1 ; P2 ; P3 ; P4 Þ
nents indicated in the residual matrix. This rather sim-
ple process will be illustrated below in the examples. Assuming that the groups P2, P3, P4 are ‘‘essentially
constant,’’ the P-space can be reduced to a simple
relationship P0 ¼ f (P1), that is, the value of Newton
Case Study I: Leuenberger (1979,1983) number Np at any point in the process is a function of
the specific amount of granulating liquid.
This example is based on the groundbreaking studies Up to this point, all the considerations were rather
conducted by Leuenberger at the University of Basel theoretical. From the theory of modeling, we know
and Sandoz AG.[43,105–108] that the above dimensional groups are functionally
The Relevance List in Table 1 reflects certain related. The form of this functional relationship f,
assumptions used to simplify the model, namely, that however, can be established only through experiments.
there are short-range interactions only and no viscosity Leuenberger and his group have empirically estab-
factor (and therefore, no Reynolds number). lished that the characteristic (that is, relative to the
Why do we have to consider the gravitational con- batch size) amount of binder liquid required to
stant? Well, imagine the same process to be done on reach a desired end-point (as expressed by the absolute
the moon—would you expect any difference? value of Np and, by proxy, in terms of net power

Table 2 The dimensional matrix for case study I

Water–Zeta

Core matrix Residual matrix

q d n P s Vb g H
Mass (M) 1 0 0 1 1 0 0 0
Length (L) 3 1 0 2 0 3 1 1
Time (T) 0 0 1 3 0 0 2 0
Wet Granulation: End-Point Determination and Scale-Up 4089

Table 3 The transformed dimensional matrix for case study I

Unity matrix Residual matrix

q d n P s Vb g H
M 1 0 0 1 1 0 0 0
3M þ L 0 1 0 5 3 3 1 1
T 0 0 1 3 0 0 2 0

consumption DP) is ‘‘scale-up invariable,’’ that is, In 2001, Holm, Schaefer, and Larsen[78] have
independent of the batch size (Fig. 11), thus specifying applied the Leuenberger method to study various pro-
the functional dependence f and establishing rational cessing factors and their effect on the correlation
basis for granulation scale-up. between power consumption and granule growth. They
Experiments with five different planetary mixers have found that such a correlation did indeed exist but
with batch sizes ranging from 3.75 kg to 60 kg showed was dependent, as expected, on the impeller design, the
that, if the binder is mixed in as a dry powder and then impeller speed, and the type of binder. The conclusion
liquid is added at a constant rate proportional to the was that it was possible to control the liquid addition
batch size, the ratio of the granulation liquid quantity by the level detection method whereby the liquid
to a batch size is constant. This was shown for non- addition is stopped at a predetermined level of power
viscous binders. consumption. An alternative approach involves an
The ratio of quantity of granulating liquid to batch inflection point (peak of the signal derivative with
size at the inflection point of power vs. time curve is respect to time).
constant irrespective of batch size and type of machine. Different vessel and blade geometry will contribute
Moreover, for a constant rate of low viscosity binder to the differences in absolute values of the signals.
addition proportional to the batch size, the rate of However, the signal profile of a given granulate com-
change (slope or time derivative) of torque or power position in a high shear mixer is very similar to the
consumption curve is linearly related to the batch size one obtained in a planetary mixer.
for a wide spectrum of high shear and planetary mix- For accuracy, in power number Np calculations, the
ers. In other words, the process end-point, as determ- power of the load on the impeller rather than the mixer
ined in a certain region of the curve, is a practically motor should be used. Before attempting to use dimen-
proven scale-up parameter for moving the product sional analysis, one has to measure/estimate power
from laboratory to production mixers of different sizes losses for empty bowl or dry stage mixing. Unlike
and manufacturers. power consumption of the impeller (based on torque
As we have indicated before, for any desired end- measurements), the baseline for motor power con-
point, the power consumption will be proportional to sumption does not stay constant and changes signifi-
the Newton power number, at a constant mixer speed. cantly with load on the impeller, mixer condition, or
The Leuenberger’s ideas relating to the use of power motor efficiency. This may present inherent difficulties
consumption for wet granulation end-point determi- in using power meters instead of torque. Torque, of
nation were tested and implemented by numerous course, is directly proportional to power drawn by
researchers.[9,33,37,41,43] the impeller (the power number can be determined

Table 4 Dimensionless P groupsa

P group Expression Definition
1 5 3
P0 P/(r  d  n ) ¼ Np Newton (Power) number
P1 s/(r1  d3  n0)
q/(rVp) Specific amount of liquid
Vp  volume of particles
Water–Zeta

q ¼ binder addition rate

t ¼ binder addition time
P2 Vb/(r0  d3  n0)
(Vp/Vb)1 Fractional particle volume
P3 g/(r0  d1  n2) ¼ Fr1 Froude Number
0 1 0
P4 H/(r  d  n ) ¼ H/d Ratio of lengths
a
Formed from the matrix in Table 3.
 4090 Wet Granulation: End-Point Determination and Scale-Up

20 [M L1 T1] column. Evidently, it was assumed that

the mass and volume could be adequately represented
in the relevance list by the density and powder height
16 in a semicylindrical vessel of a known diameter.
Np

The residual matrix (Table 7) contains four

columns; therefore four dimensionless P groups
12
(numbers) will be formed, in accordance with the
P-theorem (7 variables  3 dimensions ¼ 4 dimen-
8 sionless groups).
7 12 17 22 27 Table 8 lists the resulting groups; they correspond to
Granulating liquid quantity Newton power, Reynolds, and Froude numbers, and
(% of batch size) the ratio of characteristic lengths.
Fig. 11 Newton power number as a function of the charac-
Under the assumed condition of dynamic similarity,
teristic liquid quantity. (Adapted from Ref.[43].) from the dimensional analysis theory, it follows that
P0 ¼ f (P1, P2, P3,), and, therefore, Np ¼ f (Re,
Fr, H/d).
When corrections for gross vortexing, geometric
from the torque and speed measurements) and has a dissimilarities, and powder bed height variation were
relatively constant baseline. made, data from all mixers (Fielder PMA 25, 100
and 600 L) correlated to the extent that allows predic-
tions of the optimum end-point conditions. The linear
Case Study II: Landin et al. (1996) regression of Newton number (power) on the product
of adjusted Reynolds number, Froude number, and
Scale-up in fixed bowl mixer-granulators has been the Geometric number (in log/log domain) yields
studied by Rowe and Cliff’s group[46] using the clas- (Fig. 12) an equation of the form:
sical dimensionless numbers of Newton (power),
Reynolds, and Froude to predict the end-point in log10 Np ¼ a log10 ðRe  Fr  H=dÞ þ b
geometrically similar high-shear Fielder PMA 25,
100, and 600 L machines.
The relevance list (Table 5) included power con- where b ¼ 796 and a ¼ 0.732.
sumption of the impeller (as a response) and six factor Theoretically, in such a representation of the granu-
quantities: impeller diameter, impeller speed, vessel lation process, a slope a ¼ 1 would signify a true
height, specific density and dynamic viscosity of the laminar flow whereby a slope significantly less than
wet mass, and the gravitational constant. 1 or approaching 0 would indicate turbulence. Thus,
Note that dynamic viscosity has replaced the binder one would expect planetary mixers to have a slope clo-
amount and bowl volume of the Leuenberger’s rel- ser to 1 compared to that of high shear granulators.
evance list, thus making it applicable to viscous binders However, the results described here and in subsequent
and allowing long-range particle interactions respon- studies do not show a clear difference between
sible for friction. the slopes of regression for planetary and high-shear
The dimensional matrix for Case Study II (Table 6) mixers.
is different from Table 2: the columns for mass [M] However, the correlation coefficient of 0.7854 for
and bowl volume [L3] are replaced by a viscosity the final curve fitting effort indicates the presence of
many unexplained outlier points. One of the possible
concerns was an inherent error in measuring the height
Table 5 Relevance list for case study II (Landin et al., 1996) of the powder bed from the wet mass density.
Quantity Symbol Units Dimensions In a subsequent communication[47] it was shown
that, in order to maintain geometric similarity, it is
Power consumption P Watt M L2 T3
important to keep the batch size proportional to the
Specific density r kg/m3 M L3 bowl shape.
Water–Zeta

Blade diameter d m L Another concern is the interpretation of data from

Blade speed n rev/sec T1 mixer torque rheometer that was used to assess the vis-
Dynamic viscosity Z Pa sec M L1 T1 cosity of wet granulation. The torque values obtained
Gravitational g m/sec2 L T2
from the rheometer were labeled ‘‘wet mass consist-
constant ency’’ and were used instead of dynamic viscosity to
calculate Reynolds numbers. It was shown[45,66] that
Bowl height H m L
such torque values are proportional to kinematic
Wet Granulation: End-Point Determination and Scale-Up 4091

Table 6 The dimensional matrix for case study II

Core matrix Residual matrix

q d n P g g H
Mass (M) 1 0 0 1 1 0 0
Length (L) 3 1 0 2 1 1 1
Time (T) 0 0 1 3 1 2 0

viscosity n ¼ Z/r rather than dynamic viscosity Z Dimensional analysis and application of the Buck-
required to compute Reynolds numbers. The degree ingham theorem lead to four dimensionless quantities
of proportionality between n and Z was found to be that adequately describe the process: Ne, CRe, Fr,
formulation dependent. and h/d. As before, a relationship of the form
Consequently, it was prudent to acknowledge that
the above regression equation is not dimensionless Np ¼ 10b ðCRe  Fr  rR3b =mÞa ; or
because for all practical purposes, the Reynolds num-
log10 Np ¼ a  log10 ðCRe  Fe  rR3b =mÞ þ b
ber Re was replaced by CRe, what the authors called
a ‘‘pseudo Reynolds number’’ with the dimensions
[L3 T]. This predicament did not deter a plethora of was postulated and the constants a and b (slope and
other studies in the same line of reasoning to be pub- intercept in a log–log domain) were found empirically
lished in recent years. Note that this pseudo Reynolds (b ¼ 2.46 and a ¼ -0.872) with a good correlation
(>0.92) between the observed and predicted numbers
number has a physical meaning: it is a reciprocal of
(Fig. 13). Radius of the bowl Rb cubed was used to
volumetric flow rate.
represent the bowl volume Vb. The graph indicates
a collection of end-points produced with different
Case Study III: Faure et al. (1998) mixers and different processing factors.
It was noted that the above equation could be inter-
The same approach was applied to planetary Hobart preted to indicate that
AE240 mixer with two interchangeable bowls, 5 and
8.5 L.[49] Assuming the absence of chemical reaction DP
Z  d2  Vm =Vb
and heat transfer, the following relevance list for the
wet granulation process was suggested (Table 9): the net power consumption of the impeller varies
One difference from Table 5 of the previous study is directly with the fill ratio, wet mass viscosity, and the
the use of net power DP that was defined as motor surface swept by the blades (proportional to d2).
power consumption under load minus the dry blending Wet masses produced at the same end-point
baseline level. (regardless of bowl and batch size, impeller speed,
An assumption was made that a motor drive speed and moisture content) have been consistently shown
is proportional to the impeller blade speed. Another to result in the same final dry granule size distribution,
consideration was that the ratio of characteristic bulk density, flow, and mechanical strength.
lengths h/d is proportional to (and, therefore, can be
replaced by) a fill ratio Vm/Vb, which was, in turn,
shown to be proportional to (and therefore, could be Case Study IV: Landin et al. (1999)
replaced in the final equation by) the quantity m/
(r  d3). This is a preferred method of representing Following the methodology developed in the previous
a fill ratio because the wet mass m is easier to measure Case Study using the same assumptions, this study was
than the height of the granulation bed in the bowl. also performed on planetary mixers Collette MP20,

Table 7 The transformed dimensional matrix for case study II

Water–Zeta

Unity matrix Residual matrix

q d n P g g H
M 1 0 0 1 1 0 0
3M þ L 0 1 0 5 2 1 1
T 0 0 1 3 1 2 0
 4092 Wet Granulation: End-Point Determination and Scale-Up

Table 8 Dimensionless P groupsa

P group Expression Definition
P0 P/(r1  d5  n3) ¼ Np Newton (Power) number
P1 Z/(r1  d2  n1) ¼ Re1 Reynolds number
P2 g/(r0  d1  n2) ¼ Fr1 Froude number
0 1 0
P3 H/(r  d  n ) ¼ H/d Geometric number
(ratio of characteristic lengths)
a
Formed from the matrix in Table 7.

MP90, and MPH 200.[48] The relevance list and dimen- different wall adhesion and lid interference that was
sional matrix were the same as before, and torque partially relieved by using a Polytetrafluoroethylene
measurements from torque rheometer were again used (PTFE) lining.
to represent kinematic viscosity (instead of dynamic The end result of the dimensional analysis and
viscosity) in Reynold numbers. experimental work was, again, a regression equation
Fig. 14 represents the resulting regression line (Fig. 15) of the form

log10 Np ¼ a  log10 ðCRe  Fr  rR3b =mÞ þ b

log10 Np ¼ a  log10 ðCRe  Fr  rR3b =mÞ þ b
for the combined results from three mixers with bowl
sizes 20, 90, and 200 L showed a pretty good fit to data The regression coefficient was r2 > 0.88 using the data
(r2 > 0.95). The values for the slope and intercept from the 8, 25 and 75-L bowls with PTFE lining, and
were found to be: a ¼ 0.68, b ¼ 1280. Data from the 600-L bowl that did not require the lining. The
two other mixers with bowl sizes 5 and 40 L produces slope was found to be a ¼ 0.926, and the intercept
lines that were significantly different from the first set b ¼ 3.758.
of mixers. The authors explained this difference by
an assumption of ‘‘different flow patterns’’ in the
two groups of mixers.
Case Study VI: Hutin et al. (2004)

Case Study V: Faure et al. (1999) In this study, the foregoing methodology of dimen-
sional analysis was applied to a kneading process of
This study was done on Collette Gral Mixers (8, 25, 75, drug-cyclodextrin complexation.[109] Aoustin kneader
and 600 L) and followed the accepted—and by now, with dual Z blades was instrumented for torque mea-
standard—methodology developed earlier.[52] The surements and multiple runs were made at two scales
problem with the scale-up in the Gral mixers was the (2.5 and 5 L).
lack of geometric similitude: there was significant The relevance list for this study (Table 10) differs
‘‘distortion factor’’ between the bowl geometries at from those discussed previously by addition of blade
different scales. In addition, the researchers had to take length as one of the crucial factors affecting the
into account the lack of dynamic similitude because of process.
Introduction of the blade length, after the proper
operations with the dimensional matrix, creates
100 PMA 25 another dimensionless quantity, namely, d/l, so that
PMA 100 the resulting regression equation has the form of
10 PMA 600
Np

1 Np ¼ bðCRe  Fr  h=d  d=lÞa

0.1
Water–Zeta

100 1000 10000 100000 Experiments showed that the model fits data remark-
Re.Fr.H/d ably well (r2 > 0.99).
Fig. 12 Case study II. Regression lines of the Newton power Unfortunately, the Pharmaceutical Technology
number on the product of Reynolds number, Froude Num- journal does not grant permissions to reproduce
ber, and the length ratio for three different Fielder mixers. individual graphs; therefore, an interested reader is
(Reprinted from Ref.[46], copyright 1996, with permission from referred to the source article to see the regression lines
Elsevier.) from this study.
Wet Granulation: End-Point Determination and Scale-Up 4093

Table 9 Relevance list for case study VI (Hutin et al., 2004)

Quantity Symbol Units Dimensions
Net power DP Watt M L2 T
3
Wet mass bulk or specific density r kg/m3 M L
3
Impeller radius (or diameter) d m L
Impeller speed n rev/sec T
1
Granulation dynamic viscosity Z Pa sec M L
1 T
1
Gravitational constant g m/sec2 L T
2
Height of granulation bed in the bowl h m L

PRACTICAL CONSIDERATIONS FOR ‘‘end-point parameters’’ listed above with the processing
END-POINT DETERMINATION variables in terms of net motor power consumption
AND SCALE-UP DPm ¼ (Pe
Po) or net impeller power consumption
DPi ¼ 2p  (te
to)  n, where n is the impeller
How to Determine an End-Point? speed [dimension T
1].
Once the desired end-point is determined, it can be
A wet granulation end-point should be defined empiri- reproduced by stopping the batch at the same level
cally in terms of wet mass density and viscosity, PSD, of net power consumption DP (for the same mixer, for-
flowability or tableting parameters (e.g., capping com- mulation, speed, batch size, and amount/rate of granu-
pression). lating liquid). To account for changes in any of these
It is advisable to run a trial batch at a fixed speed variables, you have to compute the Newton power
and with a predetermined method of binder addition number Np for the desired end-point:
(for example, add water continuously at a fixed rate
to a dry mix with a water-soluble binding agent).
Np ¼ DP=ðr  n3  d5 Þ
Before adding the liquid, measure the baseline level
of motor power consumption Po or impeller torque to
at the dry mix stage. In other words, if you have established an end-point in
During the batch, stop the process frequently terms of some net impeller or motor power DP and
to take samples and, for each sample, note the end- would like to reproduce this end-point on the same
point values of power consumption Pe or impeller mixer at a different speed or wet mass density, calcu-
torque te. For each of these ‘‘end-points,’’ measure late Newton power number Np from the given Net
the resulting wet mass density r. As a result, you Impeller power DP, impeller speed n, blade radius d,
will be able to obtain some data that will relate the and wet mass density r (assuming the same batch size),

Water–Zeta

Fig. 13 Regression graph of Case Study

III. The Reynolds number Re was, in
fact, a dimensional ‘‘pseudo Reynolds
number’’ CRe. Data from a dual bowl
Hobart AE240 planetary mixer. (Repro-
duced from Ref.[49].)
 4094 Wet Granulation: End-Point Determination and Scale-Up

Fig. 14 Regression graph of Case Study

IV. The Reynolds Number Re was, in
fact, a dimensional ‘‘pseudo Reynolds’’
number CRe. Data from Collette
MP20, MP90, and MPH200 planetary
mixers. (Reproduced from Ref.[48].)

and then recalculate the target DP with the changed relating torque and dynamic viscosity (note: the corre-
values of speed n or wet mass density r. lation coefficient j can be established empirically by
Wet mass viscosity Z can be calculated from Net mixing a material with a known dynamic viscosity,
Impeller power DP, blade radius d, and impeller speed e.g., water). Alternatively, you can use impeller torque
n, using the following equations: t as a measure of kinematic viscosity and use it to
obtain a non-dimensionless ‘‘pseudo-Reynolds’’ num-
DP ¼ 2p  Dt  n ber, based on the so-called ‘‘mix consistency’’ measure,
that is, the end-point torque, as described in the case
Z ¼ j  Dt=ðn  d3 Þ studies.
Fill Ratio h/d can be calculated from a powder
where Dt is the net torque required to move wet mass, weight, granulating liquid density (1000 kg/m3 for
n is the speed of the impeller, d is the blade radius or water), rate of liquid addition, time interval for liquid
diameter, and j is mixer specific ‘‘viscosity factor’’ addition, and bowl volume Vb. The calculations are
performed using the idea that the fill ratio h/d (wet
mass height to blade diameter) is proportional to
V/Vb, and wet mass volume V can be computed as
2.5

V ¼ m=r
2.0
Log10 Np

Table 10 Relevance list for Hutin et al. (2004)

1.5
Quantity Symbol Units Dimensions
600 L Power P Watt M L2 T3
75 L + PTFE insert
1.0 25 L + PTFE insert consumption
8 L + PTFE insert Specific density r kg/m3 M L3
600 L
Other bowls Blade radius d M L
Water–Zeta

0.5 Blade speed n rev/sec T1

1.4 2.0 2.6 3.2
Log10 (ψRe·Fr·ρRb3/m) Dynamic viscosity Z Pa sec M L1 T1
Gravitational g m/sec 2
L T2
Fig. 15 Regression graph of Case Study V. Data from constant
Collette Gral 8, 25, 75, and 600 liter mixer-granulators. (Rep- Powder bed height h M L
rinted from Ref.[52], copyright 1999, with permission from
Blade length l M L
Elsevier.)
Wet Granulation: End-Point Determination and Scale-Up 4095

where m is the mass (weight) of the wet mass and r is H Bowl height (m); dimensional
the wet mass density. units [L]
Now, the weight of the wet mass is computed as the l Blade length (m); dimensional
weight of powder plus the weight of added granulating units [L]
liquid. The latter, of course, is calculated from the n Impeller speed (rev/sec);
rate and duration of the liquid addition and the liquid dimensional units [T1]
density. P Power required by the impeller
Finally, following the examples discussed in the or motor (W ¼ J/sec); dimen-
case studies, you can combine the results obtained at sional units [M L2 T5]
different end-points of the test batch or from different Rb Radius of the bowl (m); dimen-
batches or mixer scales (assuming geometrical simi- sional units [L]
larity). q Binder liquid addition rate
Given wet mass density r, wet mass viscosity Z, fill s Amount of granulating liquid
ratio h/d
m  Vb/r, setup speed n, and blade added per unit time (kg);
radius or diameter d, you can calculate the Reynolds dimensional units [M]
number Re (or the ‘‘pseudo-Reynolds’’ number) and t Binder addition time (sec);
the Froude number Fr. Then you can estimate the dimensional units [T]
slope ‘‘a’’ and intercept ‘‘b’’ of the regression equation Vp Particle volume (m3); dimen-
sional units [L3]
Np ¼ b  ðRe  Fr  h=dÞa Vm Wet mass volume (m3); dimen-
sional units [L3]
or Vb Bowl volume (m3); dimensional
units [L3]
log Np ¼ log b þ a  logðRe  Fr  h=dÞ w Wet mass; dimensional units
[M]
And, inversely, once the regression line is established, r Specific density of particles
you can calculate Newton power number Np (which (kg/m3); dimensional units [M
is the target quantity for scale-up) and net power DP L3]
(which can be observed in real time as a true indicator n ¼ Z/r Kinematic viscosity (m2/sec);
of the target end-point) for any point on the line. dimensional units [L2 T1]
Z Dynamic viscosity (Pa sec);
dimensional units [M L1 T1]
t Torque (N-m); dimensional
ACKNOWLEDGMENT
units [M L2 T2]. End-point
torque values were described
Selected excerpts and figures from M. Levin, ‘‘Granu-
as ‘‘wet mass consistency’’
lation: End-Point Theory, Instrumentation, and Scale-
numbers. Note: torque has the
Up, Education Anytime, CD-ROM Short Course,
same dimensions as work or
AAPS 1999’’ are reprinted with permission.
energy
j ¼ Z  n Dimensionless ‘‘viscosity fac-
 d3/Dt tor’’ relating net torque Dt
APPENDIX and dynamic viscosity Z
Fr ¼ n2  d/g Froude number. It relates the
List of Symbols and Dimensions inertial stress to the gravi-
tational force per unit area act-
a, b Slope and intercept of a ing on the material. It is a ratio
regression equation of the centrifugal force to the
d Impeller (blade) diameter or gravitational force
radius (m); dimensional units Np ¼ P/(r Newton (power) number. It
Water–Zeta

[L]  n3  d 5 ) relates the drag force acting

g Gravitational constant (m/ on a unit area of the impeller
sec2); dimensional units [L T2] and the inertial stress
h Height of granulation bed in Re ¼ d2 Reynolds number. It relates the
the bowl (m); dimensional units  n  r/Z inertial force to the viscous
[L] force
 4096 Wet Granulation: End-Point Determination and Scale-Up

CRe ¼ d2
n ‘‘Pseudo Reynolds number’’ 11. Schaefer, T.; Bak, H.H.; Jaegerskou, A.; Kristensen, A.;
Svensson, J.R.; Holm, P.; Kristensen, H.G. Granulation

r/t (m3/s); dimensional units [L3 in different types of high speed mixers. Part 1: effects of
T]. Note: this variable physi- process variables and up-scaling. Pharm. Ind. 1986, 48,
cally is a reciprocal of volume 1083.
12. Schaefer, T.; Bak, H.H.; Jaegerskou, A.; Kristensen, A.;
flow rate Svensson, J.R.; Holm, P.; Kristensen, H.G. Granulation
Ga ¼ Re2/Fr Galileo number in different types of high speed mixers. Part 2: comparison
between mixers. Pharm. Ind. 1987, 49, 297–304.
13. Cliff, M.J. Granulation end-point and automated process
control of mixer-granulators: Part 1. Pharm. Tech. 1990,
4, 112–132.
14. Cliff, M.J. Granulation end-point and automated process
control of mixer-granulators: Part 2. Pharm. Tech. 1990,
5, 38–44.
ARTICLES OF FURTHER INTEREST 15. Emori, H.; Sakuraba, Y.; Takahashi, K.; Nishihata, T.;
Mayumi, T. Prospective validation of high-shear wet
granulation process by wet granule sieving method. II.
Electrical Power Systems for Pharmaceutical Utility of wet granule sieving method. Drug Dev. Ind.
Equipment, p. 1482. Pharm. 1997, 23 (2), 203–215.
Fluid Bed Processes for Forming Functional 16. Corvari, V.; Fry, W.C.; Seibert, W.L.; Augsburger, L.
Instrumentation of a high-shear mixer: evaluation and
Particles, p. 1773. comparison of a new capacitive sensor, a watt meter,
Fractal Geometry in Pharmaceutical and and a strain-gage torque sensor for wet granulation.
Biological Applications, p. 1791. Pharm. Res. 1992, 9 (12), 1525–1533.
17. Corvari, V.; Fry, W.C.; Seibert, W.L.; Augsburger, L. Wet
Roller Compaction Technology for the granulation end-point detection in a high shear mixer
Pharmaceutical Industry, p. 3159. instrumented with a capacitive sensor and a strain gaged
Scale-Up and Post Approval Changes (SUPAC), torque sensor. AAPS Meeting, 1992.
18. Fry, W.C.; Stagner, W.C.; Wichman, K.C. Computer-
p. 3188. interfaced capacitive sensor for monitoring the
Tablet Press Instrumentation, p. 3684. granulation process 1: granulation monitor design and
application. J. Pharm. Sci. 1984, 73, 420–421.
19. Fry, W.C.; Stagner, W.C.; Wichman, K.C. Computer-
interfaced capacitive sensor for monitoring the granu-
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45. Landin, M.; Rowe, R.C.; York, P. Characterization of wet wet granulation’s. Pharm. Tech. Int. 1990, 2, 50–64.
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46. Landin, M.; York, P.; Cliff, M.J.; Rowe, R.C.; Wigmore, neural network to granulation scale-up. Powder Technol.
A.J. The effect of batch size on scale-up of pharmaceutical 1997, 90 (2), 153–159.
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Pharm. 1996, 133, 127–131. mixer during wet-mass granulation. Chem. Eng. Sci.
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M.J. A methodology for the optimization of wet granu- determination of a high shear granulation process. Int. J.
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Water–Zeta

50. Faure, A.; Grimsey, I.M.; Rowe, R.C.; York, P.; Cliff, Eur. 2003, 15 (6), 26–27.
M.J. Importance of wet mass consistency in the control 73. Rudd, D. The use of acoustic monitoring for the control
of wet granulation by mechanical agitation: a demon- and scale-up of a tablet granulation process. J. Proc. Anal.
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1432. 74. Miwa, A.; Toshio Yajima, T.; Itai, S. Prediction of suitable
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M.J. Applicability of a scale-up methodology for wet Pharm. 2000, 195 (1–2), 81–92.
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wet granulation process. Pharm. Dev. Technol. 2001, 6 (2), control. Int. J. Pharm. 1997, 156 (1), 109–117.
181–192. 95. Badawy, S.I.F.; Menning, M.; Gorko, M.A.; Gilbert, D.L.
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World Health Organization (WHO): Global
Harmonization of Requirements for
Medicinal Products
Juhana E. Idanpaan-Heikkila
World Health Organization, Geneva, Switzerland

INTRODUCTION Naming the biotechnology-derived substances

and products received increasing attention. Prevention
The World Health Organization (WHO) is an inter- of INNs from using trademarks has now been
governmental specialized agency of the United Nations strengthened in collaboration with the World Intellec-
with 193 member states. Regarding pharmaceutical tual Property Organization (WIPO). A new point of
and biological products, WHO has the global mandate collaboration was the protection of INNs against
to develop, establish, and promote quality, safety, and misuse of Internet domain names.[4]
efficacy standards and codes of good practices for all
medicinal products. Vigorous harmonization of quality,
safety, efficacy, and nomenclature requirements on a
worldwide basis has continued in 1998–2000 as the INTERNATIONAL PHARMACOPOEIA—
explicit responsibility of WHO. The content of the consti- 50 YEARS OLD
tutional mandate, expertise involved, working practices
and rules, consultative processes, and role of the gov- Publication of the International Pharmacopoeia has
erning bodies regarding WHO’s normative work.[1] This continued now for 50 years. Its role has been to fulfill
review gives an update on ongoing harmonization a need in developing countries where use of less tech-
activities since 1998 and outlines the new harmonization nically advanced tests using simple instrumentation is
initiatives. common practice for specific substances and prepara-
tions. Publishing monographs for finished products
has also been useful. In 1999, the Expert Committee
recommended that less-advanced methods should be
INTERNATIONAL NON-PROPRIETARY developed in parallel with modern analytical tech-
NAMES (INNs) niques because in some developing countries, more
sophisticated methods could be useful.[4] Volume 5 of
The International Non-proprietary Names (INNs) the International Pharmacopoeia is in print. WHO
identifies pharmaceutical substances by unique, glo- has completed monographs for several antimalarials
bally recognized names. A single internationally recog- including artemisin derivatives. Basic tests for pharma-
nized name for an active drug substance is a starting ceutical substances, medicinal plant materials, and
point for its pharmacopeial monograph, safe prescrib- dosage forms (including 345 tests for substances, 208
ing and dispensing, and easy communication among for dosage forms, and four for plant materials) were
scientists and health professionals worldwide. The published in 1998.[5] The 69 International Infrared
INN is for common use without restrictions (except Reference Spectra (IIRS) have been made available
as a trademark); it is distinctive in sound and spelling from the WHO Collaborating Centre for Chemical
and not too long. In 1998–1999, WHO published more Reference Substances (Kungens Kurva, Sweden). The
than 270 new INNs in its quarterly publication, WHO list of reference substances and infrared reference
Water–Zeta

Drug Information[2] and issued a revised Guidelines spectra is updated regularly and is now available
on the Use of International Non-proprietary Names on the World Wide Web (http://www.who.int/dmp/
(INN) for Pharmaceutical Substances.[3] The revision irintro.htm).
of the procedure included the introduction of a service A revised guideline on Good Practices for National
fee, improvements in Procedures to raise objections Pharmaceutical Control Laboratories (GPCL) was
to proposed INNs, and the replacement of a recom- published in 1999. It takes into account guidances
mended INN. from ISO 17025, EN 45001, and OECD–GLP and
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-100000446
Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved. 4099
 4100 World Health Organization (WHO): Global Harmonization of Requirements for Medicinal Products

recommendations published by the Pharmaceutical countries as a model instrument to exchange infor-

Inspection Convention (PIC).[4] mation among authorities in importing and exporting
countries. In 1999, WHO completed a model certificate
of analysis to be used for trade of starting materials
WHO GMP AND QUALITY ASSURANCE intended for drug manufacturing, and manufacturers
of pharmaceutical substances, excipients, and medicinal
In 1998, WHO began a special project to assist in products.[4]
implementing of GMP in member states with limited
resources. Countries interested in upgrading their
Comparator Product System for
manufacturing have been identified, and training
Equivalence Testing
material on basic GMP principles has been prepared
together with training modules for advanced GMP
The use of generic pharmaceutical products has
topics and inspections. Several regional training work-
increased in many countries. Consequently, drug regu-
shops for trainers in GMP have been organized,
latory authorities are increasingly involved in the
together with missions to concerned countries.[4]
assessment of the scientific documentation submitted
The following guidelines have been completed:
GMP for Sterile Pharmaceutical Products, Guideline for marketing approval of generic medicinal products.
WHO has stated that the quality, safety, and efficacy
on Pre-Approval Inspection (before granting market-
of a generic product must comply with the same
ing authorization), Quality System for National
requirements applied to the innovator.[10] Conse-
GMP Inspectorate, and General Aspects of Packaging
quently, the therapeutic equivalency and interchange-
Pharmaceuticals.[4]
ability with the innovator must be proven with valid
In 1999, WHO published a report on the control
comparative in vivo bioequivalence studies. To assist
and safe trade of starting materials for pharmaceutical
both manufacturers and drug regulators in this,
products in English, French, Spanish, Russian, Chi-
nese, and Arabic, based on recommendations from a WHO has developed a list of international comparator
products. The information on comparator products
meeting of experts in Geneva in May 1998.[6] The meet-
was collected first from drug regulatory agencies, and
ing was inspired by several recent incidents of contami-
a provisional list of candidate products was consulted
nation with highly toxic solvent diethylene glycol.
with the innovator companies concerned. The compa-
Because of their increasing use worldwide, plant
nies were requested to designate the market and trade-
materials used in over-the-counter preparations, home
mark that best represented the quality, manufacturing,
remedies, or as raw materials for pharmaceutical
and labeling of the product. The current list thus far
preparations are receiving more and more attention.
In 1998, WHO published a book, Quality Control contains 147 products from the WHO Model List of
Essential Drugs, complemented with detailed instruc-
Methods for Medicinal Plant Materials to fulfill the
tions for use and a decision tree for special situa-
needs of quality control laboratories and to provide a
tions.[11] The list will be updated periodically by WHO.
basis for the development of national standards.[7]
A compendium of WHO guidelines and related
materials entitled Quality Assurance of Pharmaceuti-
cals, Volume 1, was published[8] and the next volume THE WHO MODEL LIST OF
is in print. ESSENTIAL DRUGS

The Model List of Essential Drugs was created in 1975,

Counterfeit Drugs and the most recent assessment and updating of the list
took place in 1997 and 1999.[12,13] The following addi-
In 1999, WHO published guidelines on the detection tions to the list were made including aciclovir for use
and prevention of counterfeit and substandard pro- in herpes infections; amoxicillin þ clavulanic acid for
ducts.[9] Vigilance and reporting of counterfeit products the treatment of infections resistant to the production
to WHO were enhanced by setting up a liaison officer of b-lactamase; dextrometorphan for the treatment of
network among drug regulatory authorities and WHO. cough; ephedrine for treatment of hypotension in
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spinal anesthesia during delivery; imipenem þ cilastatin

for the treatment of Pseudomonas; ipratropium bromide
THE WHO CERTIFICATION SCHEME for the treatment of asthma; metformin for the treatment
of non-insulin-dependent diabetes; triclabendazole for the
The WHO Certification Scheme on the Quality of treatment of liver and lung flukes; and nevirapine and
Pharmaceutical Products Moving in International zidovudine to reduce or prevent mother-to-child trans-
Commerce[1] was used in increasing the number of mission of HIV infection. The value of lipid-lowering
World Health Organization (WHO): Global Harmonization of Requirements for Medicinal Products 4101

agents was discussed; however, because of their high The Assembly appealed to the pharmaceutical
costs, they were not included in the list, with a recommen- industry, health professional and consumer organiza-
dation that each country make its own decisions for use tions, and other interested parties to promote the
in high-risk patient populations.[12] formulation and use of good information practices
consistent with the principles of the WHO Ethical
Sale and Promotion of Medical Products Criteria for Medicinal Drug Promotion; monitor and
on the Internet report problem cases and aspects of cross-border
advertising, promotion, and sale of medical products
The Internet is a rapidly expanding medium that has using the Internet; and maintain legal and ethical
many uses and great potential for disseminating and standards in these activities.
obtaining information regarding a variety of subjects WHO was requested to encourage the international
such as new treatments, institutions offering care, community to formulate self-regulatory guidelines for
and medical products available. Individuals can get good information practices and to develop a model
this information more quickly and conveniently than guide for member states. The guide should educate
in traditional ways, such as in medical journals and and instruct people using the Internet on how to best
textbooks. Organizations, pharmaceutical companies, obtain reliable, independent, and compatible infor-
and individuals post and exchange information about mation on medical products through this medium.
their aims, activities, or products and sometimes offer The WHO model guide, Medical Products and the
to sell their products, including medical products. Internet, was published in 1999.[17]
However, have the products been approved for mar-
keting? Are they safe, effective, properly labeled, and
of good quality? Reported cases show that dangerous, Biologicals
harmful, unreliable, low-quality, substandard, fake,
counterfeit, and mislabeled products have been avail- WHO’s normative work for biological substances used
able on the Internet.[14] in medicinal products, including vaccines, blood pro-
The World Health Assembly (the annual summit of ducts, and diagnostic procedures, has resulted in a
Ministries of Health of 193 member states of WHO) number of globally applicable specifications, guide-
recognized in May 1997 the increasing use of electronic lines, and guiding principles. In 1999, WHO published
communication by the general public to shop and guidelines for the production and quality control of
gather information on health education, diseases, treat- synthetic peptide vaccines for human use to ensure
ments, and medical products.[15] The Assembly was their consistent safety and efficacy.[18] The guidelines
concerned about reported cases of inappropriate use address control of starting materials including back-
of the Internet, which had caused a potential hazard ground data on the synthesis of the peptide of interest
for public health, and decided to convene a WHO ad and control of the manufacturing process and the final
hoc working group to formulate recommendations product. WHO requirements for tick-borne encepha-
for action. The working group met in September litis vaccine (inactivated) were formulated to take into
1997 in Geneva and consisted of representatives of account current manufacturing practices and control
health and drug regulatory authorities, consumer procedures in place and to give guidance on how these
groups, professional associations, the pharmaceutical practices and procedures could be updated.[18]
industry, experts in legal and ethical matters, market- Requirements for hepatitis B vaccines made by recom-
ing, and other interested parties. The group reviewed binant DNA techniques have been amended to reflect
the situation and made several recommendations recent developments in assay methodology, as well as
regarding cross-border advertising, promotion, and for specifications for Hemophilus type b conjugate vac-
sale of medical products. Based on these recommenda- cines. Under development were guidelines for standar-
tions, in May 1998[16] the World Health Assembly dization and calibration of cytokine immunoassays,
urged all member states to review existing legislation, and requirements for tetravalent dengue vaccine (live)
regulations, and guidelines to ensure that they are and oral poliomyelitis vaccine.[18]
applicable and adequate to cover the promotion and In 1999, WHO published requirements for pro-
sale of medical products over the Internet. Member duction and control of Haemophilus influenzae type b
Water–Zeta

states were asked to set up monitoring and surveillance (Hib) conjugate vaccine and the acellular perussis
systems for the Internet. Collaboration was recom- component of monovalent or combined vaccines.
mended among countries to identify difficult cases The requirements for oral polio vaccine (OPV) were
and disseminate information through WHO. The also revised in 1999, with several additions. WHO
availability of scientific, validated information to published new and replacement International Standards
consumers by competent health authorities was and Reference Materials covering a wide range of
determined to be necessary. products.[19]
 4102 World Health Organization (WHO): Global Harmonization of Requirements for Medicinal Products

WHO, ICH, AND REGIONAL HARMONIZATION REFERENCES

The role of WHO as an observer of the ICH and its 1. WHO and the harmonization of regulatory requirements
for medicinal products. In Encyclopedia of Pharma-
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member states have been described in detail.[1] suppl. 1.
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of experts from the International Generic Pharmaceutical 3. WHO. Guidelines on the Use of International Non-
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line for good manufacturing practices for active phar- Medicinal Plant Materials and Dosage Forms; WHO:
maceutical ingredients based on preliminary work by Geneva, 1998.
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(PIC/S). Australia, China, andIndia, known for their Geneva, 1998.
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bat Counterfeit Drugs; WHO/EDM/QSM/99.1, WHO:
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WHO regional adviser for pharmaceuticals from each lence testing. Drug Information 1999, 13, 158–162.
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of the six WHO regions to coodinate among ICH mation 1998, 12, 22–35.
CTD working groups and drug regulatory authorities 13. WHO. model list (revised in November 1999). Drug Infor-
WHO regions.[20] This is an important development mation 1999, 13.
14. Idänpään-Heikkilä, J.E. Marketing pharmaceuticals on the
because the CTD will undoubtedly have an impact internet. In Medicines on the Internet. Communication
on new drug applications in non-ICH countries and from the Industry to the Patients, Nordic Council on Med-
industries. The CTD expert working groups succeeded icines; 52, NLN: Stockholm, 2000; 63–67.
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cover all four climatic zones.[21,22] 25–34.

X-Ray Powder Diffractometry
Raj Suryanarayanan
College of Pharmacy, University of Minnesota, Minneapolis, Minnesota, U.S.A.

Suneel Rastogi
Forest Laboratories, Inc., Inwood, New York, U.S.A.

INTRODUCTION incident on a crystalline sample at an angle y. Diffraction

will occur if:
X-rays are electromagnetic radiation lying between
ultraviolet and gamma rays in the electromagnetic nl ¼ 2; d sin y ð1Þ
spectrum. The wavelength of the X-ray region is con-
sidered to be between 0.01 and 100 Å.[1] where d ¼ distance between the successive planes in
There are two broad applications of X-rays in the the crystal lattice, expressed in Å, and n ¼ order of
characterization of materials: (i) X-ray spectrometry reflection (an integer).
and (ii) X-ray diffractometry. The former technique X-ray patterns can be obtained using either a
is used for chemical analysis and has found only lim- powder diffractometer or a camera. Currently, diffract-
ited use in the characterization of pharmaceuticals. ometers find widespread use in the analysis of
On the other hand, X-ray diffractometry, by providing pharmaceutical solids. The technique is usually non-
a means for the study of the structure of crystalline destructive in nature. The theory and operation of
materials, is extensively used to characterize pharma- powder diffractometers is outside the scope of this
ceutical solids. There are two principal applications discussion, but these topics have received excellent
of X-ray diffractometry. X-ray crystallography is coverage elsewhere.[1–4] Instead, the discussion will be
concerned with the structure determination of crystal- restricted to the applications of X-ray powder diffrac-
line phases. Single crystals are usually used for this tometry (XRD) in the analysis of pharmaceutical
purpose. On the other hand, in X-ray powder diffrac- solids. The United States Pharmacopeia provides
tometry, the sample is usually in the form of a powder. a brief but comprehensive introduction to X-ray dif-
X-ray powder diffractometry is recognized as a power- fractometry.[5] The use of XRD in the physical charac-
ful technique for the identification of crystalline terization of pharmaceutical solids[6] and in the
phases. The technique can also be used for the quanti- characterization of controlled release delivery systems
tative analyses of solids. This article will be restricted have been discussed earlier.[7]
to the principles and applications of X-ray powder
diffractometry (XRD) in the characterization of phar-
maceutical solids.
Diffraction is a scattering phenomenon. When QUALITATIVE ANALYSIS
X-rays are incident on crystalline solids, they are
scattered in all directions. In some of these directions, Since the X-ray diffraction pattern of every crystalline
the scattered beams are completely in phase and form of a compound is unique, the technique is widely
reinforce one another to form the diffracted beams.[1,2] used for the identification and characterization of solid
The Bragg law describes the conditions under which phases. XRD is the technique of choice to identify
this would occur. It is assumed that a perfectly parallel different polymorphic forms of a compound
and monochromatic X-ray beam, of wavelength l, is (Fig. 1).[8] It can also be used to identify the solvated
and unsolvated (anhydrous) forms of a compound,
Water–Zeta

provided their lattice structures are different. The tech-

nique can also reveal differences in the crystallinity of
compounds. The XRD pattern of an amorphous (non-
Some parts of this article have been reproduced from the chapter, crystalline) compound will consist of one or more
‘‘X-ray Powder Diffractometry,’’ written by R. Suryanarayanan in
‘‘Physical Characterization of Pharmaceutical Solids,’’ edited by broad diffuse halos (Fig. 2A).[9]
H. Brittain. This was Vol. 70 of Drugs and Pharmaceutical Sciences, The United States Pharmacopeia contains the XRD
Editor: J. Swarbrick, Marcel Dekker, Inc., New York, 1995. patterns of two anhydrous forms (Form 1 and Form 2)
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-100200014
Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved. 4103
 4104 X-Ray Powder Diffractometry

Fig. 1 X-ray powder diffraction (XRD) patterns of the

different solid forms of AG337. (From Ref.[8].)

Fig. 2 (A) XRD patterns of (a) crystalline and (b) amorph-

of ampicillin, ampicillin trihydrate and amorphous ous sucrose. (B) XRD patterns of the physical mixtures
anhydrous ampicillin.[5] The powder patterns of these of crystalline and amorphous sucrose. The sucrose content
ranged between 0 and 5 wt%. The 12.7 and 13.1 2y peaks
four solid phases reveal pronounced differences, which
were used for the quantitative analyses. [(B) Reproduced
can form the basis of their identification.
from Ref.[9].]
XRD was a powerful tool for the characterization
of the different solid phases of AG337 (a 5-substituted
quinazolinone; dihydrochloride salt). In addition demonstrated the unique ability of XRD for the identi-
to several polymorphic forms of the anhydrate fication of (a) an anhydrous compound existing in
(C14H12N4OS
2HCl), the compound existed as a both crystalline and amorphous states, (b) different
hemihydrate (C14H12N4OS
2HCl
0.5H2O), a mono- polymorphic forms of the anhydrate, (c) the existence
hydrate (C14H12N4OS
2HCl
H2O), and a dihydrate of solvates, where the solvent of crystallization is
(C14H12N4OS
2HCl
2H2O).[8] The hemihydrate, the water (hydrate) or an organic solvent (in this case, tert-
dihydrate, and one of the polymorphic forms of the butylamine), and (d) polymorphism in the hydrate.
anhydrate (Id) had identical lattice structures (Fig. 1).
While the amorphous and crystalline solid forms of
sucrose can be readily distinguished by XRD (Fig. 2A), Reference Diffraction Patterns
the technique is also capable of revealing the presence
of small amounts of crystalline sucrose in the presence The International Centre for Diffraction Data (ICDD,
of amorphous sucrose.[9] This is evident from the XRD Newtown Square, PA) maintains a collection of single-
patterns of physical mixtures of amorphous and crys- phase X-ray powder patterns.[11] There are separate
talline sucrose wherein the crystalline sucrose content listings of inorganic and organic compounds. The
ranged between 1 and 5 wt% (Fig. 2B). X-ray diffraction data of ibuprofen, as a representative
Water–Zeta

Six crystalline solid phases of fluprednisolone and example is given here (Table 1).
an amorphous phase were characterized using XRD, The card pattern contains the Powder Diffraction
IR spectroscopy, and differential scanning calo- File (PDF) number (Region 1); quality mark of the
rimetry.[10] Three of these six crystalline phases were data (Region 2); the chemical formula and the speci-
anhydrous, two were monohydrates, and one was a tert- men name (Region 3); the experimental conditions
butylamine disolvate. The differences in the powder under which the powder pattern was obtained and
patterns of the phases were readily evident. This study the source of the data (Region 4); physical data that
X-Ray Powder Diffractometry 4105

Table 1 The card pattern of ibuprofen

include crystallographic system, space group, lattice an ‘‘O’’ mark is assigned. Patterns that do not meet the
parameters, and interaxial angles (Region 5); general criteria for ‘‘ ,’’ ‘‘i,’’ or ‘‘O’’ are left blank. When the
comments, Crystal Data cell (if different from that powder pattern is calculated from structural param-
reported in Region 5); Pearson Symbol Code (PSC), eters, the pattern is marked ‘‘C.’’ Extensive details
Merck Index number, etc. (Region 6); and a table of about the quality mark guidelines can be found in
interplanar spacings, relative intensities, and the Miller ICDD publications.[12]
indices (Region 7). Another important database for pharmaceuticals
There are also several indicators of the quality of is the Cambridge Structural Database (CSD). Main-
the data. The highest quality data are given . To qual- tained by the Cambridge Crystallographic Data Centre,
ify for this mark, the chemistry of the compound must CSD is a compilation of single-phase data containing
be well characterized. The intensities of the X-ray lines structural information including unit cell parameters,
must be measured objectively and instrumentally and crystal system, and space group. The structural infor-
there must be no unindexed, space group extinct or mation can be used to calculate powder diffraction
impurity lines. Lines with d-spacings of 2.50 Å must patterns. The ICDD and the Cambridge Crystallo-
retain at least three significant digits after the decimal graphic Data Centre have a cooperative program. This
point. To qualify for the ‘‘i’’ mark, there can be a allows the ICDD to calculate powder patterns for
Water–Zeta

maximum of two unindexed, space group extinct or inclusion in the Powder Diffraction File, from the data
impurity lines provided none of these belong to the recorded in the CSD.
strongest eight. Again there must be no serious system-
atic errors and lines with d-spacings 2.00 Å must Phase Identification in Solid Dosage Forms
retain at least three significant digits after the decimal
point. If the data are of low precision or if the data In addition to the active ingredient, solid dosage forms
are due to a poorly characterized or multiphase system, usually contain one or more excipients. In such powder
 4106 X-Ray Powder Diffractometry

mixtures, each crystalline phase produces its pattern

independently of the other constituents in the mixture.
Thus, the unique advantage of XRD is that it com-
bines absolute specificity with a high degree of accu-
racy. Shell pioneered the use of XRD in the
characterization of pharmaceutical dosage forms.[13]
The recent advances in instrumentation and software
have substantially enhanced the utility of the tech-
nique. XRD was used to simultaneously identify the
three active ingredients, acetaminophen, aspirin, and
caffeine, in a commercially available tablet formulation
(ExcedrinÕ). The three active ingredients together
constituted 83 wt% of the formulation.[14] The XRD
pattern contained numerous peaks in the angular Fig. 4 (a) The residual XRD pattern after proportional
range of 7–37 2y (Fig. 3d). In an effort to identify subtraction of acetaminophen and aspirin XRD patterns
the components in the dosage form, the XRD patterns from that of the powdered tablet formulation. (b) The
of acetaminophen, aspirin, and caffeine were obtained XRD pattern of caffeine powder. (From Ref.[14].)
(Fig. 3a–c). Acetaminophen could be readily identified
by two unique lines with d-spacings of 6.48 and
4.92 Å (2y values of 13.65 and 18.00 , respectively). the literature.[14] When the subtracted profile (Fig. 4a)
At these 2y values, the XRD patterns of aspirin and was compared with that of caffeine (Fig. 4b), the high
caffeine contained no peaks. Similarly, aspirin could intensity peaks of caffeine at 11.70, 26.25, and 26.85
be readily identified by two unique lines with 2y were readily discernible. However, an amorphous
d-spacings of 11.54 and 3.95 Å (2y values of 7.65 and halo was observed over the angular range of 18–23
22.45 , respectively). Caffeine had an intense line with 2y. The formulation contained numerous excipients
a d-spacing of 7.55 Å (2y value of 11.70 ) and two including microcrystalline cellulose, hydroxypropyl
intense lines with d-spacings of 3.39 and 3.31Å (2y methylcellulose, and hydroxypropyl cellulose. The
values of 26.25 and 26.85 , respectively). Since at these XRD pattern of microcrystalline cellulose exhibited a
2y values, peaks due to aspirin and acetaminophen broad halo over the angular range of 18–25 2y
occurred, unambiguous identification of caffeine was (Fig. 5c). While amorphous halos were also observed
not possible. An added complication was that caffeine in the XRD patterns of hydroxypropyl methylcellulose
constituted only 8.9 wt% of the formulation. (Fig. 5b) and hydroxypropyl cellulose (Fig. 5a), their
In order to identify caffeine, the XRD patterns of angular range did not match that of the formulation
acetaminophen and aspirin were selectively subtracted
from the XRD pattern of the formulation. The details
of the pattern subtraction procedure are described in
Water–Zeta

Fig. 5 (a) The XRD patterns of hydroxypropyl cellulose

powder. (b) The XRD pattern of hydroxypropyl methyl-
Fig. 3 (a) The XRD pattern of acetaminophen powder. (b) cellulose powder. (c) The XRD pattern of microcrystalline
The XRD pattern of aspirin powder. (c) The XRD pattern of cellulose powder. (d) The residual XRD pattern after pro-
caffeine powder. (d) The XRD pattern of a powdered tablet portional subtraction of acetaminophen and aspirin XRD
formulation containing acetaminophen, aspirin, and caffeine patterns from that of the powdered tablet formulation.
(ExcedrinÕ). (From Ref.[14].) (From Ref.[14].)
X-Ray Powder Diffractometry 4107

can be analyzed directly with minimal or no sample

pretreatment. There is no need to extract the active
ingredient from the dosage form, since it can be usually
characterized in presence of the excipient(s). The tech-
nique will permit simultaneous identification and
quantification of more than one active ingredient in
formulations. Finally, XRD can provide quantitative
information about the degree of crystallinity.
When theophylline monohydrate was dehydrated, it
formed a metastable anhydrous phase, which then
transformed to the stable anhydrate.[15] The XRD pat-
terns of the two anhydrate phases and the monohy-
drate were sufficiently different so that all three of
Fig. 6 (a) The XRD pattern of chlordiazepoxide hydro- them could be simultaneously identified in a sample
chloride powder. (b) The XRD pattern of a powder mixture (Fig. 7A). Anhydrous theophylline was granulated
of chlordiazepoxide hydrochloride (5 wt%) and microcrystalline with an aqueous solution of PVP. During the wet-
cellulose (95 wt%). (c) The XRD pattern of microcrystalline massing stage, the anhydrate transformed to the
cellulose powder. (d) The residual XRD pattern after pro- monohydrate. When the granules were dried, there was
portional subtraction of the microcrystalline cellulose XRD dehydration resulting in the formation of the meta-
pattern from that of the physical mixture (b). The full scale in stable anhydrate, which then transformed to the stable
this case is different from that of the other three XRD patterns.
(From Ref.[14].)

(Fig. 5d). Therefore, microcrystalline cellulose is likely

to be the major contributor to the observed halo. Thus,
XRD not only permitted simultaneous identification of
all the active ingredients in the dosage form, but it also
provided information about the excipients in the for-
mulation.
The authors also evaluated the sensitivity of the
method, using chlordiazepoxide HCl (hereafter referred
to as chlordiazepoxide) as the model compound.[14]
While chlordiazepoxide was crystalline (Fig. 6a), micro-
crystalline cellulose exhibited a broad amorphous halo
(Fig. 6c). Identifying chlordiazepoxide was no problem
so long as its weight fraction was 0.10. When the drug
weight fraction was decreased to 0.05, its presence was
not readily discernible (Fig. 6b). Using the pattern
subtraction technique, the XRD pattern of microcrys-
talline cellulose was subtracted from the XRD pattern
of the drug-microcrystalline cellulose mixture (Fig. 6d).
This permitted ready identification of chlordiazepoxide
(compare Fig. 6d with 6a).

Phase Transitions Induced

During Processing

The components of a dosage form, both active ingredi-

Fig. 7 (A) XRD patterns of theophylline phases. M, A, and
Water–Zeta

ents and excipients, can undergo a variety of physical

A refer to theophylline monohydrate, stable anhydrous
transformations during pharmaceutical processing
theophylline, and metastable anhydrous theophylline respec-
and storage. These include polymorphic transforma- tively. (B) Phase transitions during the drying of theophylline
tions, alterations in crystallinity, and changes in the granules. PVP was the granulating agent. The integrated
state and degree of hydration. XRD is well-suited for intensities of the 8.9, 9.4, and 7.0 2y peaks unique to M,
analyses of such transformations, provided the ana- A , and A respectively were simultaneously monitored as a
lytes are crystalline. In most cases, the dosage forms function of the drying time. (From Ref.[15].)
 4108 X-Ray Powder Diffractometry

anhydrate. All three phases were simultaneously

monitored by XRD (Fig. 7B).

DEGREE OF CRYSTALLINITY

Solids may be either crystalline or non-crystalline. The

crystalline state is characterized by a perfectly ordered
lattice and the non-crystalline (amorphous) state is
characterized by a disordered lattice. These represent
two extremes of lattice order and intermediate states
are possible. The term degree of crystallinity is useful
in attempts to quantify these intermediate states of lat-
tice order.
XRD is widely used to determine the degree of crys-
tallinity of pharmaceuticals. The procedure developed
by Hermans and Weidinger[16] is based on three
assumptions. First, it must be possible to demarcate
and measure the crystalline intensity (Ic) and amorph-
ous intensity (Ia) from the powder pattern. Usually, the Fig. 8 A plot of the integrated amorphous intensity (Iam) as
integrated line intensity (area under the curve), rather a function of the integrated crystalline intensity (Ic) for lac-
than the peak intensity (peak height), is measured. tose samples milled for various time periods. (From Ref.[17].)
Second, there is a proportionality between the
experimentally measured crystalline intensity and the
years, several techniques have been developed to quan-
crystalline fraction (xc) in the sample. Finally, a pro-
tify low levels of an amorphous compound in presence
portionality exists between the experimentally mea-
of its crystalline counterpart. Using sucrose as a model
sured amorphous intensity and the amorphous
compound, an XRD method was developed to detect
fraction (xa) in the sample. The degree of crystallinity
and to quantify crystalline sucrose when it occurred
(or percent crystallinity), xcr, is given by the expression,
as a mixture with amorphous sucrose.[9] The XRD
patterns of amorphous and crystalline sucrose were
Ic 100 presented earlier (Fig. 2). Standards consisting of
xcr ¼ ð2Þ
Ic þ qIpa amorphous and crystalline sucrose were prepared
wherein the crystalline sucrose content ranged between
1 and 5 wt%. The sum of the background subtracted
where p and q are proportionality constants. The integrated intensities of the 12.7 2y (6.94 Å) and
values of Ia and Ic can be determined for samples of 13.1 2y (6.73 Å) peaks of sucrose (these peaks have
varying degrees of crystallinity. A plot of the measured been marked with an asterisk in Fig. 2) were linearly
values of Ia against those of Ic will result in a straight related to the weight percent of sucrose (Fig. 9).
line, and the intercepts on the y- and x-axes will pro- The limits of detection and quantitation of crystal-
vide the intensity values of the 100% amorphous and line sucrose were determined to be 0.9 and 1.8 wt%
100% crystalline materials, respectively. This method respectively. Water sorption and FT–Raman spec-
was used by Nakai et al.[17] to estimate the degree of troscopy also appear to be very sensitive with detection
crystallinity of lactose that had been milled for various possible down to levels of 1 wt%.[20,21]
time periods (Fig. 8). If the value of (q/p) is known, the
degree of crystallinity of an unknown sample can be
calculated from the experimentally determined values
of Ic and Ia. The degree of crystallinity of microcrystal- PHASE QUANTIFICATION
line cellulose and digoxin milled for various time peri- (QUANTITATIVE ANALYSIS)
Water–Zeta

ods were also determined.[18,19] The problems and

limitations of this method have been discussed in the The Direct Method (No Internal Standard)
literature.[6]
A number of drugs and excipients exist in the Alexander and Klug developed the theoretical basis of
amorphous state. However, the physical instability of the quantitative analyses of powder mixtures.[3]
amorphous compounds can lead to their crystallization Although a powder mixture may be composed of sev-
resulting in partially crystalline materials. In recent eral components, it can be regarded as being composed
X-Ray Powder Diffractometry 4109

This approach was used to determine the weight

fractions of anhydrous carbamazepine and carbamaze-
pine dihydrate when they occurred as mixtures.[22]
Based on the mass attenuation coefficients of the anhy-
drate and the dihydrate, the intensity ratios [Ii1/(Ii1)0]
were calculated as a function of the anhydrate content
in the mixture (the line in Fig. 10). These were in good
agreement with the experimentally obtained values of
[Ii1/(Ii1)0].
A simpler system is a two component mixture,
where m1 ¼ mM 
. This occurs when the analyte and the
matrix have the same molecular formula, as in poly-
morphic mixtures. In such cases, Eq. (4) reduces to:
Fig. 9 Plot of the sum of the intensities of the 12.7 and 13.1
2y peaks as a function of the weight percent of crystalline
Ii1
sucrose in mixtures of amorphous and crystalline sucrose. ¼ x1 ð5Þ
(From Ref.[9].) ðIi1 Þ0

Therefore a plot of the relative intensity as a func-

of just two components: Component 1 (which is the
tion of the analyte weight fraction would be linear.
unknown) and the sum of the other components
This approach was used to determine the weight
(which is designated as the matrix). The relationship
fractions of anhydrous a-carbamazepine and b-
between the intensity of peak i of the unknown compo-
carbamazepine when they occurred as mixtures.[23]
nent (Ii1) and its weight fraction (x1) in the mixture is
While studying the polymorphism of 1,2-dihydro-6-
given by the equation:
neopentyl-2-oxonicotinic acid, Chao and Vail[24] used
XRD to quantify Form I in mixtures of Forms I and
Kx1 II. They estimated that Form I levels as low as 0.5 wt%
Ii1 ¼ ð3Þ
r1 ½x1 ðm1  mM Þ þ mM  can be determined by this technique. Similarly the

where K is a constant, r1 is the density of Component

1, and m1 and mM 
are the mass attenuation coefficients
of Components 1 and the matrix respectively. The
mass attenuation coefficient of a substance is the
weighted average of the mass attenuation coefficients
of its constituent elements.
Let us first consider a two-component mixture,
where m1 6¼ mM
. One example is a mixture of the anhy-
drous and hydrated forms of a compound. It is first
necessary to determine the intensity of peak i of a sam-
ple consisting of only the analyte [(Ii1)0]. Next, Eq. (3)
is modified so that the relative intensity of the XRD
peak of Component 1 [expressed as [(Ii1)/(Ii1)0] is
expressed as:

Ii1 x1 m1
¼ ð4Þ
ðIi1 Þ0 x1 ðm1  mM Þ þ mM


It is possible to calculate the mass attenuation coef-

ficients of the analyte and the matrix based on their
Water–Zeta

chemical compositions.[2] This enables the calculation

of the relative intensity [Ii1/(Ii1)0] as a function of the
Fig. 10 The relative intensity of the 6.78 Å line of anhydrous
weight fraction of the analyte in the mixture and thus carbamazepine (b-form) as a function of its weight fraction in
generate a theoretical curve. This eliminates the need binary mixtures of anhydrous carbamazepine (b-form) and
for the preparation of experimental standard curves. carbamazepine dihydrate. The line is based on theoretical
An added advantage of this approach is that there is values, while the data points are experimental measurements.
no requirement of an internal standard. (From Ref.[22].)
 4110 X-Ray Powder Diffractometry

a-inosine content in a mixture consisting of a-and When dealing with organic pharmaceutical systems,
b-inosine was achieved with a detection limit of 0.4 wt% an organic internal standard is preferred. Since the
for a-inosine.[25] XRD patterns of organic compounds usually contain
numerous lines, it might not be possible to identify lines
unique to the analyte and the internal standard that
Internal Standard Method are completely separated from one another. Therefore,
inorganic compounds such as corundum (a-Al2O3),
In this method, an internal standard is added, and its silicon, lithium fluoride, and zinc oxide are often
weight fraction is maintained constant in all the mix- used as internal standards. Methenamine, an organic
tures. This method was used to simultaneously quan- compound, has several of the properties desired in an
tify the S(þ)-enantiomer and the racemic compound internal standard.[13] However, it has not found wide-
of ibuprofen.[26] spread use in the analyses of pharmaceuticals.

Selection of Internal Standard Whole Powder Pattern Analyses

Success in quantitative XRD may hinge on the selec- In the pharmaceutical community, quantitative analy-
tion of an appropriate Internal Standard. The follow- ses has conventionally been based on the intensity of a
ing properties are desired in an internal standard.[13] characteristic peak of the analyte. It is now recognized
(i) The compound must have high crystal symmetry that phase quantification will be more accurate if
so that strong, but few, diffraction peaks are produced. it is based on the entire powder pattern.[27,28] This
(ii) The high-intensity lines in the analyte and the inter- forms the basis for the whole-powder-pattern analyses
nal standard that are to be used for quantitative pur- method developed in the last few decades. Of the avail-
poses should not overlap with one another, but they able methods, the Rietveld method is deemed the most
should be close to each other. (iii) The density of the powerful since it is based on structural parameters.
internal standard should be close to that of the system This is a whole-pattern fitting least-squares refinement
ingredients so that homogeneous mixtures can be pre- technique that has also been extensively used for
pared for analysis. (iv) The internal standard must crystal structure refinement and to determine the size
be chemically stable in the experimental system. and strain of crystallites.

Table 2 Measured composition of carbamazepine mixtures and the residuals obtained by the Rietveld analysis
Actual composition of carbamazepine (wt %) Measured composition of carbamazepine (wt %)

a b Dihydrate Lithium fluoride a b Dihydrate Lithium fluoride Rawp (%)

80 20 79 21 20.0
80 20 79 21 22.7
80 20 80 20 20.7
79.6 0.4 20 78 1 21 21.5
79.2 0.8 20 80 1 19 21.4
79.2 0.8 20 78 2 21 20.9
79.2 0.8 20 78 2 21 21.4
0.4 79.6 20 <1 79 20 20.7
0.4 79.6 20 <1 79 21 21.3
0.4 79.6 20 <1 78 22 20.9
0.8 79.2 20 1 78 21 20.9
0.4 78.8 0.8 20 <1 77 1 21 21.3
0.8 78.8 0.4 20 <1 78 1 21 20.1
Water–Zeta

4 72 4 20 3 73 5 19 15.5
4 72 4 20 2 71 7 19 26.9
4 72 4 20 2 71 7 19 26.9
4 40 36 20 2 41 36 20 22.3
a
Weighted profile residual.
(From Ref.[29].)
X-Ray Powder Diffractometry 4111

Simultaneous quantitative analyses, of complex The phase transitions of theophylline monohydrate

pharmaceutical mixtures, has been accomplished by were discussed earlier in the section dealing with Phase
the Rietveld method.[29] Using lithium fluoride as an Transitions Induced During Processing (Fig. 7).
internal standard, mixtures consisting of anhydrous High temperature XRD was used to investigate the
b-carbamazepine, anhydrous a-carbamazepine, and dehydration of theophylline monohydrate (Fig. 11).
carbamazepine dihydrate were subjected to quantita- This technique revealed that dehydration resulted in
tive analyses (Table 2). When the analyte concentration a metastable anhydrous phase, which then transformed
was high (20%), the method was accurate, with a to the stable anhydrate.[30] Theophylline has been in
relative error <5%. However, when the analyte con- widespread use for many years and its solid-state
centration was low (<5%), the method lacked accuracy. properties have been the subject of numerous investi-
gations. Without high temperature XRD, it is unlikely
that this metastable phase would have been identified.
NON-AMBIENT XRD
Kinetics of Solid-State Reactions
Conventionally, XRD patterns are obtained at room
temperature under ambient conditions. Variable tem-
Solid-state reactions in pharmaceuticals can be broadly
perature XRD is a technique where XRD patterns
classified into chemical decompositions and physical
are obtained while the sample is subjected to a con-
transformations. The chemical decomposition of drugs
trolled temperature program. It is also possible to con-
has received adequate attention in the pharmaceutical
trol the environment and maintain the sample under
literature. The active ingredient in a solid dosage form
the desired relative humidity. Recent advances in
can also undergo a variety of physical transformations
commercially available instrumentation have greatly
including polymorphic transitions, alterations in
facilitated the study of pharmaceutical systems under
degree of crystallinity, and alterations in degree of sol-
non-ambient conditions. Experiments can be carried out
vation. These transformations can affect pharmaceuti-
both at elevated temperatures and under subambient
cally important properties such as solubility, stability,
conditions.
powder flow, and tabletting behavior, which ultimately
can cause variations in the performance of the pro-
High Temperature XRD duct. There are strict pharmacopeial standards, which
govern not only the chemical purity of drugs and
In the characterization of pharmaceutical systems, excipients, but also the drug content in dosage
high temperature XRD has been used extensively. forms. On the other hand, very few compounds have

Water–Zeta

Fig. 11 High temperature XRD of theophylline monohydrate. XRD patterns were obtained at the temperatures indicated in the
figure. The ‘‘ ’’, ‘‘þ,’’ and ‘‘’’ marks indicate peaks unique to metastable anhydrous theophylline (referred to as A in Fig. 7),
stable anhydrous theophylline (A in Fig. 7), and theophylline monohydrate (M in Fig. 7) respectively. (Adapted from Ref.[30].)
 4112 X-Ray Powder Diffractometry

pharmacopeial standards that govern their physical enantiomers were identical, the racemic compound
form. However, it is increasingly recognized that the exhibited a different powder pattern. XRD permitted
physical form of a compound and physical transforma- the simultaneous quantification of the disappearance
tions in a dosage form can profoundly affect product of the enantiomers and the subsequent appearance of
performance. An added complication is that conven- the racemic compound. The rate of disappearance
tional analytical techniques such as HPLC, which are of the enantiomers followed a diffusion-controlled
used for monitoring chemical decomposition reactions reaction model. During the kinetic experiment, the
are unsuitable to monitor solid-state physical transfor- sum of the weight fractions of the enantiomers and
mations. This is because of their inability to distinguish the racemic compound progressively decreased from
between the different solid forms of a compound. Both an initial value of unity, suggesting the formation of
chemical decomposition as well as physical transitions an intermediate non-crystalline phase. This was con-
can occur simultaneously during the processing or storage firmed by a steady increase in the background counts
of solid formulations. Therefore, it would be ideal if an in the powder pattern.
analytical technique can simultaneously monitor both The sweetener aspartame exists as a hemihydrate
types of transformations. XRD is one such technique. (C14H18N2O5
0.5H2O; ASH), under ambient con-
Conventionally, thermoanalytical techniques, specifi- ditions. When heated, ASH converted to aspartame
cally differential scanning calorimetry and thermogravi- anhydrate (ASA), which on further heating decom-
metric analysis, have been used to investigate the kinetics posed to form a diketopiperazine (DKP) derivative.[33]
of solid-state reactions. These techniques have several The XRD patterns of ASH, ASA, and DKP showed
limitations. They do not unambiguously identify crystal- pronounced differences (Fig. 12). XRD was used to
line phases and are incapable of providing quantitative simultaneously quantify the (i) disappearance of ASH
information about the degree of crystallinity. Inter- and appearance of ASA in the first reaction and (ii)
mediate phases, if formed, may not be readily identified. disappearance of ASA and appearance of DKP in
The techniques may therefore be unsuitable for discern- the second reaction. For studying the kinetics of the
ing the reaction mechanism. When two or more gaseous first reaction, the peaks unique to ASH at 15.9 and
products are simultaneously evolved, thermal analysis 16.4 2y and the 17.1 2y peak of ASA were used. For
alone is not very helpful in the study of reaction kinetics. the second reaction, the sum of the integrated intensi-
In such cases, thermal analysis should be used in con- ties of the peaks at 10.2, 11.0, and 11.8 2y of ASA
junction with other techniques for evolved gas analysis and the 13.0 2y peak of DKP were used. While the
(e.g. infrared spectroscopy, mass spectroscopy, or dehydration of ASH appeared to follow first-order
gas chromatography). It is often realized that solid-state kinetics, the cyclization of ASA was a nucleation-
reactions take place through complex mechanisms controlled process. Figs. 13 and 14 contain the
involving intermediates and the same end product is dehydration and cyclization data at 118 and 180 C
obtained through multiple pathways. In such cases, respectively. Since the concentrations of the crystalline
thermoanalytical techniques yield complex profiles reactant as well as the product were simultaneously
because of overlapping thermal events. Often, it is diffi- monitored, mass balance calculations of the crystalline
cult to separate the reaction steps and study the kinetics phases were possible at each time point. The reaction
of the individual steps. XRD does not suffer from these
limitations and can therefore serve as an excellent
complement to thermoanalytical techniques. Recent
advances in instrumentation have enabled simultaneous
and independent control of the temperature and the
water vapor pressure in the sample chamber.
The dehydration kinetics of theophylline monohy-
drate (C7H8N4O2
H2O) and ampicillin trihydrate
(C16H19N3O4S
3H2O) were studied by XRD.[31]
Dehydration of theophylline monohydrate resulted in
a crystalline anhydrous phase, while the ampicillin trihy-
drate formed an amorphous anhydrate. In the case of
Water–Zeta

theophylline, simultaneous quantification of both the

monohydrate and the anhydrate was possible. By carrying
out the reaction at several temperatures, the activation Fig. 12 XRD patterns of aspartame hemihydrate (ASH),
energy for the dehydration reaction was determined. aspartame anhydrate (ASA) and diketopiperazine derivative
The enantiomers of pseudoephedrine were observed (DKP). ‘‘ ,’’ ‘‘’’and ‘‘þ’’ denote the peaks unique to aspar-
to react in the solid state to form the racemic tame hemihydrate, aspartame anhydrate, and diketopipera-
compound.[32] While the powder patterns of the zine derivative respectively. (From Ref.[33].)
X-Ray Powder Diffractometry 4113

Thus, irrespective of the nature of the product phase,

XRD is capable of revealing such interactions.

Low Temperature XRD

The use of a cooling accessory permits XRD patterns

to be obtained under subambient conditions. In phar-
maceutical systems, the greatest utility of the technique
is to monitor the crystallization of solutes in frozen
solutions. Conventionally, differential scanning calo-
rimetry has been the most popular technique for the
characterization of frozen systems. However, as men-
Fig. 13 Weight fractions of ASH and ASA as a function of tioned earlier, this technique has some drawbacks: (i)
time following storage of ASH at 118 C. The curves are It does not enable direct identification of crystalline
drawn only to assist in visualizing the trends. (From Ref.[33].) solid phase(s). Moreover, it is difficult to draw any
definitive conclusions about the degree of crystallinity.
(ii) The interpretation of DSC curves is very difficult if
rate constants were obtained at several temperatures, there are overlapping thermal events. Low temperature
which permitted the calculation of the activation XRD was found to be an excellent complement to dif-
energies of dehydration and cyclization from the ferential thermal analysis in the characterization of
Arrhenius plots. water–glycine–sucrose ternary systems.[34]
When sodium nafcillin solutions were frozen, the
solute did not crystallize.[35] However, annealing
Drug–Excipient Interactions caused solute crystallization. It was also possible to
monitor the amount of sodium nafcillin crystallizing
XRD is an excellent technique to study drug–excipient as a function of annealing time. XRD was also useful
interactions, provided the drug and the excipients are to characterize the behavior of buffer solutions. While
crystalline. In a solid mixture, the powder pattern of disodium hydrogen phosphate crystallized when an
each crystalline phase is produced independently aqueous solution was frozen, the presence of mono-
of the other constituents. Thus the diffraction pattern sodium hydrogen phosphate inhibited the crystalliza-
of a powder mixture will be the summation of the dif- tion of the former.
fraction patterns of the individual constituents. If the
drug–excipient interaction results in a crystalline pro- In situ freeze-drying
duct, this will be characterized by the appearance of
new peaks in the powder pattern. However, if the The low temperature stage of the X-ray powder dif-
interaction results in an amorphous product, this will fractometer was modified so that the sample chamber
become evident from the broad halos in the pattern. could be evacuated. As a result, the entire freeze-drying
process was carried out in situ in the sample chamber
of the XRD. The advantage of such a set-up is that
the alterations in the solid state during the various
Weight fraction of crystalline phase

Sum of the weight fractions

1 stages of the freeze-drying process (cooling, primary,
and secondary drying) were monitored.[36] As men-
DKP
0.8 tioned earlier, annealing of frozen aqueous solutions
of sodium nafcillin resulted in the crystallization of
0.6 sodium nafcillin hydrate (the hydrate stoichiometry
was not known). During the primary drying, there
0.4 was partial dehydration resulting in the formation of
partially crystalline sodium nafcillin hemihydrate
Water–Zeta

0.2 (Fig. 15). It was possible to simultaneously monitor

ASA
the disappearance of sodium nafcillin hydrate as well
0 as the appearance of sodium nafcillin hemihydrate
0 10 20 30 40 50
Time (min)
(Fig. 16). XRD revealed that this reaction was com-
plete in about 40 min. The technique was also used to
Fig. 14 Weight fractions of ASA and DKP as a function of monitor the phase transitions during the freeze-drying
time, following storage of ASA at 180 C. (From Ref.[33].) of mannitol.
 4114 X-Ray Powder Diffractometry

different stages of freeze-drying will not only aid in

the optimization of the freeze-drying cycle, but will
also facilitate the production of formulations that are
physically and chemically stable.

ISSUES AND CHALLENGES IN XRD

Most materials of pharmaceutical interest are organic

compounds. Jenkins has comprehensively discussed
the problems associated with the X-ray diffractometric
analysis of organic materials:[37] 1) The unit cells
of organic compounds are often large, resulting in
low angle lines (large d-spacings). The accuracy of
Fig. 15 The XRD patterns of (a) ‘‘as is’’ sodium nafcillin, d-spacing data tends to decrease as the d-spacing
(b) frozen solution of sodium nafcillin (40 wt%) annealed at
increases, particularly when the d-spacing value is
4 C for 1 h, and (c) after freeze-drying the annealed system.
The ‘‘as is’’ sodium nafcillin was a monohydrate. Annealing higher than 8 Å; 2) The unit cells are often of low sym-
is believed to result in the crystallization of a higher hydrate metry resulting in complex powder patterns; 3) Since
of sodium nafcillin (unknown stoichiometry). The freeze- organic compounds usually have low mass attenuation
dried material was partially crystalline sodium nafcillin hemi- coefficients, the X-rays can penetrate deep into the
hydrate. (From Ref.[35].) specimen leading to large beam transparency errors;
and 4) Finally, organic compounds are especially
prone to preferred orientation (preferred orientation
is discussed in the following section).
Implications in the Design of
Freeze-Dried Formulations
Sample Preparation
Most solid-state characterization studies investigate
the starting material and the freeze-dried end product. There are many different methods of sample prep-
It will be useful and relevant to study alterations in the aration. Most of the sample holders contain a cavity
solid state during the different stages of the freeze- into which the powder sample is filled, usually from
drying process, and this is enabled by low temperature the top. However, the side-loading (also referred to
XRD. The physical characterization of the phases at as side-drift) method is considered the best packing
method.[1,38] Unfortunately, many commercial sample
holders do not permit sample filling by this method.
As a result, holders have been specially fabricated for
this purpose.[23] Since organic compounds predomi-
nantly consist of atoms with low atomic numbers,
there will be significant penetration of X-rays. This
can result in peak displacement as well as broadening.
Detailed information on sample preparation is avail-
able in the literature.[4]
When only a very small amount of powder sample is
available, the use of a low-background (also referred to
as zero background) support is advisable. The support
is a quartz or silicon plate cut along a non-diffracting
crystallographic direction, which is then polished.[1]
This results in a very low background and improves
the signal-to-noise ratio and enables XRD patterns to
Water–Zeta

be obtained with extremely small amounts of sample.

Fig. 16 Real time monitoring of the phase changes during
the freeze-drying of sodium nafcillin by in situ XRD. The dis-
appearance of sodium nafcillin hydrate (‘‘Hydrate’’) and the Sources of Error
appearance of sodium nafcillin hemihydrate (‘‘Hemi-
hydrate’’) were simultaneously quantified. The curves were There are numerous sources of error in XRD. Two of
drawn to assist in visualizing the trends. (From Ref.[36].) these are discussed hereafter.
X-Ray Powder Diffractometry 4115

crystallites. When the crystallites in a sample are oriented

in some particular (non-random) manner, they are said
to exhibit preferred orientation.[1] Ideally, random orien-
tation of individual crystallites is desired. It is recognized
that preferred orientation is a commonly encountered
condition in powdered samples.[2]
Grinding the sample and thereby reducing the par-
ticle size can decrease preferred orientation. However,
grinding can cause lattice disorder (decrease in crystal-
linity) and also induce other undesirable transitions,
such as polymorphic transformations. Moreover, when
the particle size is very small, particle size induced peak
broadening can be observed. This effect, observed
when the particle size is <10,000 Å (1 mm), is simply a
consequence of the limited number of planes that are
diffracting X-rays (Fig. 18). The particle (crystallite)
size is related to the X-ray line width according to
the Scherrer Eq. (1). It is important to recognize the
fact that line broadening can be brought about by a
loss in crystallinity as well as a decrease in particle size.
Fig. 17 The structural makeup of solids. (From Ref.[1].) In the pharmaceutical literature, there is a tendency to
automatically attribute line broadening to loss in crys-
tallinity. Such a conclusion, while usually correct, can
Preferred Orientation
be occasionally erroneous.
Reliable and reproducible results can only be obtained
if the sample is prepared carefully. When a sample is Measurement of Line Intensity
packed in an X-ray holder, the distribution of crystal
orientations can be non-random, and this condition When measuring intensity, the integrated line intensity
is referred to as preferred orientation. Jenkins and (peak area) and not the maximum intensity (peak
Snyder[1] have elegantly illustrated this point in Fig. 17. height) must be measured. Variations in lattice strain
Atoms bond together, in a unique manner, to form unit and particle size can significantly influence the line
cells. If the unit cells are grouped so as to have long- shape, but their effect on the integrated intensity will
range order, a crystalline material is obtained. In the generally be minimal.[2]
absence of long-range order, the material is amorphous
(non-crystalline). A powder sample is composed of Other Sources of Error

There are numerous other sources of error that are

3.0
particularly relevant while conducting quantitative XRD
studies. These are discussed in detail in the literature.[1–3,6]

ACKNOWLEDGMENT
2.0
Line breadth [˚2θ]

We thank Mr. Rahul Surana for his assistance in the

preparation of the manuscript.

1.0
REFERENCES
Water–Zeta

1. Jenkins, R.; Snyder, R.L. Introduction to X-Ray Powder

Diffractometry; Wiley: New York, NY, 1996.
Instrumental Broadening
2. Cullity, B.D. Elements of X-Ray Diffraction, 2nd Ed.;
0 Addison-Wesley: Reading, MA, 1978.
10 20 50 100 200 500 1000 2000 5000 10000
3. Klug, H.P.; Alexander, L.E. X-Ray Diffraction Procedures
Particle dimension [Å] for Polycrystalline and Amorphous Materials, 2nd Ed.;
Wiley: New York, NY, 1974.
Fig. 18 The width of X-ray lines as a function of particle 4. Buhrke, V.E.; Jenkins, R.; Smith, D.K. A Practical Guide
dimension. (From Ref.[1].) for the Preparation of Specimens for X-Ray Fluorescence
 4116 X-Ray Powder Diffractometry

and X-Ray Diffraction Analysis; Wiley-VCH: New York, in a mixture by powder X-ray diffractometry. Pharm. Res.
NY, 1998. 1989, 6 (12), 1017–1024.
5. The United States Pharmacopeia; 24th Rev.; U.S. Pharma- 23. Suryanarayanan, R. Determination of the relative amounts
copeial Convention: Rockville, MD, 1999. of a-carbamazepine and b-carbamazepine in a mixture by
6. Suryanarayanan, R. X-Ray powder diffractometry. In Physi- powder X-ray diffractometry. Powder Diffr. 1990, 5 (3),
cal Characterization of Pharmaceutical Solids; Brittain, 155–159.
H.G., Ed.; Marcel Dekker, Inc.: New York, NY, 1995. 24. Chao, R.S.; Vail, K.C. Polymorphism of 1,2-dihydro-
7. Rastogi, S.; Suryanarayanan, R. Characterization of deliv- 6-neopentyl-2-oxonicotinic acid: characterization, intercon-
ery systems—X-ray powder diffractometry. In The Encyc- version, and quantitation. Pharm. Res. 1987, 4 (5), 429–432.
lopedia of Controlled Drug Delivery; Mathiowitz, E., 25. Doff, D.H.; Brownen, F.L.; Corrigan, O.I. Determination
Ed.; Wiley: New York, NY, 1999. of a-impurities in the b-polymorph of inosine using infrared
8. Rastogi, S.; Zamansky, I.; Roy, S.; Tyle, P.; Suryanarayanan, spectroscopy and X-ray powder diffraction. Analyst
R. Solid-state phase transitions of AG337, an antitumor (London) 1986, 111 (2), 179–182.
agent. Pharm. Dev. Technol. 1999, 4 (4), 623–632. 26. Phadnis, N.V.; Suryanarayanan, R. Simultaneous quantifi-
9. Surana, R.; Suryanarayanan, R. Quantitation of crystal- cation of an enantiomer and the racemic compound of ibu-
linity in substantially amorphous pharmaceuticals and profen by X-ray powder diffractometry. Pharm. Res. 1997,
study of crystallization kinetics by X-ray powder diffracto- 14 (9), 1176–1180.
metry. Powder Diffr. 2000, 15 (1), 2–6. 27. Anwar, J. Analysis of time-resolved powder diffraction
10. Haleblian, J.K.; Koda, R.T.; Biles, J.A. Isolation and char- data using a pattern-decomposition method with restraints.
acterization of some solid phases of fluprednisolone. J. Appl. Crystallogr. 1993, 26 (3), 413–421.
J. Pharm. Sci. 1971, 60 (10), 1485–1488. 28. Rietveld, H.M. Profile refinement method for nuclear and
11. McClune, W.F., Ed.; Powder Diffraction File, Inorganic magnetic structures. J. Appl. Crystallogr. 1969, 2, 65–71.
Phases, Organic and Organometallic Phases Search 29. Iyengar, S.S.; Phadnis, N.V.; Suryanarayanan, R. Quanti-
Manual; International Centre for Diffraction Data: tative analyses of complex pharmaceutical mixtures by the
Newtown Square, PA, 1993. rietveld method. Powder Diffr. In press.
12. Powder Diffraction file: set 43. In Inorganic and Organic 30. Phadnis, N.V.; Suryanarayanan, R. Polymorphism in anhy-
Databook; McClune, W.F., Ed.; International Centre for drous theophylline-implications for the dissolution rate
Diffraction Data: Newtown Square, PA, 1993. of theophylline tablets. J. Pharm. Sci. 1997, 86 (11), 1256–
13. Shell, J.W. X-ray and crystallographic applications in phar- 1263.
maceutical research, II.. J. Pharm. Sci. 1963, 52, 24–39. 31. Shefter, E.; Fung, H.-L.; Mok, O. Dehydration of crystal-
14. Phadnis, N.V.; Cavatur, R.K.; Suryanarayanan, R. Identifi- line theophylline monohydrate and ampicillin trihydrate.
cation of drugs in pharmaceutical dosage forms by X-ray J.Pharm. Sci. 1973, 62 (5), 791–794.
powder diffractometry. J. Pharm. Biomed. Anal. 1997, 32. Duddu, S.P.; Khin-Khin, A.; Grant, D.J.W.; Suryanarayanan,
15 (7), 929–943. R. A novel X-ray powder diffractometric method for studying
15. Tank, J. Changes in the Solid-State of Theophylline Upon the reaction between pseudoephedrine enantiomers. J. Pharm.
Aqueous Wet Granulation. M.S. thesis, Department of Phar- Sci. 1997, 86 (3), 340–345.
maceutics, University of Minnesota: Minneapolis, MN, 1997. 33. Rastogi, S.; Zakrzewski, M.; Suryanarayanan, R. Investi-
16. Hermans, P.H.; Weidinger, A. Quantitative X-ray investi- gation of solid-state reactions using variable temperature
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analysis. J. Appl. Phys. 1948, 19, 491–506. Pharm. Res. 2001, 18 (3), 267–273.
17. Nakai, Y.; Fukuoka, E.; Nakajima, S.; Morita, M. Physico- 34. Shalaev, E.Y.; Malakhov, D.V.; Kanev, A.N.; Kosyakov,
chemical properties of crystalline lactose. I. Estimation of V.I.; Tuzikov, F.V.; Varaksin, N.A.; Vavilin, V.I. Study of
the degree of crystallinity and the disorder parameter by the phase diagram water fraction of the system water–
an X-ray diffraction method. Chem. Pharm. Bull. 1982, glycine–sucrose by DTA and X-ray diffraction methods.
30 (5), 1811–1818. Thermochim. Acta 1992, 196 (1), 213–220.
18. Nakai, Y.; Fukuoka, E.; Nakajima, S.; Hasegawa, J. Crys- 35. Cavatur, R.K.; Suryanarayanan, R. Characterization of
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cellulose. Chem. Pharm. Bull. 1977, 25 (1), 96–101. diffractometry. Pharm. Res. 1998, 15 (2), 194–199.
19. Black, D.B.; Lovering, E.G. Estimation of the degree of 36. Cavatur, R.K.; Suryanarayanan, R. Characterization of
crystallinity in digoxin by X-ray and infrared methods. phase transitions during freeze-drying by in situ X-ray
J. Pharm. Pharmacol. 1977, 29 (11), 684–687. powder diffractometry. Pharm. Dev. Technol. 1998, 3 (4),
20. Saleki-Gerhardt, A.; Ahlneck, C.; Zografi, G. Assessment 579–586.
of disorder in crystalline solids. Int. J. Pharm. 1994, 101, 37. Jenkins, R. New directions in the X-ray diffraction analysis
237–247. of organic materials. Adv. X-Ray Anal. 1992, 35, 653–660.
21. Taylor, L.S.; Zografi, G. Quantitative analysis of crystallinity 38. McMurdie, H.F.; Morris, M.C.; Evans, E.H.; Paretzkin, B.;
using FT-Raman spectroscopy. Pharm. Res. 1998, 15, 755–761. Wong-Ng, W.; Hubbard, C.R. Methods of producing stan-
22. Suryanarayanan, R. Determination of the relative amounts dard X-ray diffraction powder patterns. Powder Diffr.
of anhydrous carbamazepine and carbamazepine dihydrate 1986, 1 (1), 40–43.
Water–Zeta
Zeta Potential
Luk Chiu Li
HPD Advanced Drug Delivery, Abbott Park, Illinois, U.S.A.

Youqin Tian
Alcon Laboratories, Fort Worth, Texas, U.S.A.

INTRODUCTION zeta potential (z). It can provide a measure of the net

surface charge on the particle and potential distri-
Dispersion systems represent an important class of bution at the interface. Zeta potential serves as an
pharmaceutical dosage forms such as emulsions, sus- important parameter in characterizing the electrostatic
pensions, microspheres, liposomes, and nanoparticles. interaction between particles in dispersed systems and
The medium of these systems is mainly aqueous in the properties of the dispersion as affected by this
nature and the dispersed phase can be either solid electrical phenomenon.
particles or immiscible liquid droplets. Electrical
charges are developed by several mechanisms at the
interface between the dispersed phase and the aqueous METHOD AND INSTRUMENTATION
medium.[1] The two most common mechanisms are the
ionization of surface functional groups and the specific Microelectrophoresis
adsorption of ions. These electrical charges play an
important role in determining the interaction between The most common method for determining the zeta-
particles of the dispersed phase and the resultant physi- potential is the microelectrophoretic procedure in
cal stability of the systems, particularly for those in the which the movements of individual particles under
colloidal size range. For dispersed systems which are the influence of a known electric field are followed
used as drug carriers, i.e., liposomes and microparti- microscopically. The zeta potential can be calculated
cles, their in vivo fate and therapeutic efficacy are from the electrophoretic velocity of the particles using
both affected by these surface charges. Therefore, the the Helmholtz–Smoluchowski equation,
understanding of this electrical phenomenon becomes
essential in developing these systems. Up ¼ Vp E ¼ zeZ ð1Þ
The presence of these surface charges influences an
uneven distribution of charges (ions) surrounding the where Up is the electrophoretic mobility, Vp is the elec-
particle and the development of an electrical potential trophoretic velocity, E is the electric-field strength,
(or an electrical field) between the surface and the elec- z is the zeta potential, e is the permittivity, and Z is
trically neutral bulk-solution phase of the system. The the viscosity of the medium.
surface charge and the counterions in its vicinity give It is important to realize that the observed electro-
rise to an electric double layer as shown in Fig. 1. phoretic migration is the sum of two contributions,
The double layer is divided into two parts separated one of which is the electro-osmotic flow of the medium
by a plane called the Stern layer which is located at through the cell. The glass of the sample cell generally
about a hydrated-ion radius from the surface. Usually, bears negative charges, so that a diffuse layer of
the number of ions in the Stern layer is smaller than cations exists adjacent to them. This sets up an electro-
that needed to achieve neutralization of the surface osmotic flow through the cell (with or without parti-
charge and the balance of the neutralization occurs cles). This flow has its maximum value at the center,
in a Gouy–Chapman layer outside the Stern layer. since the layer of fluid adjacent to the walls is station-
The surface potential (c0) and the Stern potential ary. The particles tracked at the center of the cell
Water–Zeta

(cd) are not readily measured experimentally; instead, therefore possess the maximum increment in velocity
the potential between a stationary fluid layer envelop- because of the electro-osmotic flow. Since there is no
ing the particle and the bulk-solution phase can be net liquid flow through the closed cell, a back-pressure
determined by measuring the mobility of the particle builds up which causes fluid flow in the reverse
in an applied electrical field. The potential between direction in the core of the cell. In the steady state,
the tightly bound surface liquid layer (shear plane) of these flows balance, giving rise to a velocity profile as
the particle and the bulk phase of the solution is called shown in Fig. 2. Therefore, the tracking of particles
Encyclopedia of Pharmaceutical Technology DOI: 10.1081/E-EPT-100200015
Copyright # 2007 by Informa Healthcare USA, Inc. All rights reserved. 4117
 4118 Zeta Potential

Gouy plane – – increases, however, the susceptibility to errors also

– + –
increases. For example, a long measurement time will
– + + + – +
Diffuse layer – + + cause convection currents generated by heating of the
– – + –
+ –+ + sample, air-bubble formation, settling of particles,
– – – Net + +
+ +
+ +– –
negative + – –
Bulk solution and electrode reactions. These potential errors can be
Shear plane + +
+ + particle– + +
+
– avoided if the measurement is taken much more
–
– + ++ +– –+ + +
+ + + + quickly without loss of accuracy. A rotating prism
Stern plane – + + + + –
method employed by the Lazer Zee Meter (Model
+
+– – + + +
+ –
+ 50,1 Pen Kem, Inc.), allows the operator to measure
Particle surface more than 10 particles simultaneously. Data are
+ Potential ψe
thereby attained more rapidly and, consequently, with
ψd
fewer time-dependent errors. Fig. 3 shows a diagram of
ξ light paths in the rotating prism method.[2] The micro-
scope is focused on the stationary layer of the cell (with
no fluid flow). Special cylindrical optics compress the
1/k Distance from
particle surface
laser beam into an illumination sheet of laser light. A
vertical adjustment is provided to vary the height so
Fig. 1 A schematic representation of the electric double that it coincides with the focal plane of the microscope.
layer around a particle with negative surface charges and A cube prism inside the microscope causes the viewed
electrical potentials surrounding the particle. image to translate at a rate proportional to the prism’s
speed of rotation. The applied electric field causes the
particle to move at an electrophoretic velocity pro-
at a location where the medium experiences no net flow portional to the mobility and the applied voltage.
becomes important in yielding an accurate electro- Adjustment of the potentiometer causes the prism to
phoretic mobility measurement. A velocity profile has rotate at a rate proportional to potentiometer voltage
been derived for the liquid contained in a cylindrical times applied voltage. When the image velocity caused
electrophoretic cell. The location with zero liquid flow by prism rotation is equal and opposite to the mean
is shown to be 14.6% of the cell diameter inside the sur- electrophoretic velocity, the cloud of particles appears
face of the capillary. The location of the surface of zero stationary. The applied voltage is therefore pro-
liquid flow in cells of rectangular cross section has also portional to the mean particle electrophoric mobility.
been determined. For a cell in which the direction of The mobility value is scaled by an appropriate
migration is relatively long compared to the width of constant in the digital meter and the computed zeta
the cell, the surface with zero liquid flow lies 21.1% potential is displayed on the 3-digit readout.
of the cell depth above the bottom and below the top In spite of a major improvement gained from the
of the working compartment. Experimentally, particles rotating prism method, there are still serious limita-
tracked at these positions of the sample cell will display tions in the use of microelectrophoresis in determining
their mobility uncomplicated by the electro-osmotic zeta potential. First, this technique cannot be applied
effect. to dense dispersions, i.e., the number density of the
The rate of particle migration can be determined by particles must be very low. This can sometimes, but
measuring with a stopwatch the time required for a not always, be remedied by filtering the sample and
particle to travel between the marks of a calibrated diluting a small amount of the original dispersion with
graticule in the microscope eyepiece. Since many parti- a large volume of filtrate. Second, it cannot be applied
cles are usually measured in order to get good statisti-
cal results, the measurement of a sample may take
30 min or even longer. As the time of measurement

Prism Galvanometer
Beam-forming
optics Chamber
Water–Zeta

Laser
Planes of 0 velocity

Vertical
adjustment

Fig. 2 The velocity profile of fluid flow in an electrophorec- Fig. 3 A diagram of light paths in the rotating prism
tic cell. method of microelectrophoretic measurement.
Zeta Potential 4119

to particles less than about 0.2 mm in diameter because zeta-potential distribution.[6] This instrument provides
they are not easily visible in the ultramicrosope, simultaneous measurements of line widths at four
though the particle-size range may be extended by different angles so that the motion of the particle can
adsorbing the colloidal particles (i.e., protein mole- be decomposed with high resolution into components
cules) on suitable carrier particles. An additional and caused by electrophoretic and diffusional motions.
unresolvable problem with very small particles is the Therefore, particle sizing can also be carried out by
blurring effect imparted by Brownian motion. Further- the same instrument simultaneously with electrophore-
more, the microelectrophoretic technique cannot be tic mobility determinations. Fig. 4 presents a schematic
used to determine mobility distributions or to separate diagram of the DELSA 440 system. Other commer-
mutlimodal mobility distributions. cially available electrophoretic analyzers using the
laser light-scattering principles are the Zetasizer 3
(Malvern Instruments), ZetaPlus (Brookhaven Instru-
Electrophoretic Light-Scattering[3–5] ments), System 3000 (PenKem, Inc.), and NICOMP
380/ZLS (Particle Sizing Systems).
Electrophoretic light-scattering is a technique which
determines the electrophoretic velocities by measuring
the Doppler shifts of scattered laser light. The electro- Electrokinetic Sonic Analysis[3,6,7]
phoretic mobility can be expressed by
When a colloidal dispersion is subject to ultrasonic
Up ¼ Dnl0 =2nE sinðy=2Þ ð2Þ waves, a density difference between the dispersed phase
and the continuous phase will cause relative motion
where Up is electrophoretic mobility, Dn is the Doppler between the particles and the surrounding fluid. This
frequency shift, l0 is the wavelength of the laser in a relative motion means that there will be a periodic
vacuum, n is the index of refraction of the medium, interchange of the charged particle surface and the
E is the electric field applied, and y is the scattering oppositely charged counter-ions in the electric double
angle. Since a He–Ne laser light has a frequency of layer. This interchange results in the development of
about 6.0
1014, a direct measurement of frequency an alternating dipole at the frequency of the applied
shifts ranging from zero to several hundred Hz may ultrasound wave. This effect is termed the ultrasound
be problematic because of the difficulty in measuring vibration potential (UVP). The UVP effect in colloids
a small difference between large quantities. However, is also referred to as the colloid vibration potential
this can be overcome by using the principle of hetero- or CVP. An effect opposite that of CVP is generated,
dyning, which measures the difference in frequency when an alternating electric field is applied to a
resulting from a reference beam and the Doppler- colloidal dispersion, creating an acoustic wave which,
shifted beam. by means of a piezoelectric device, may be transduced
When the Brownian motion of the particles is negli- into an alternating potential of a certain amplitude,
gible, the power spectrum which is generated by the depending on the dispersion and the electric double-
autocorrelation function of the scattered light has a layer properties. This is called the electrokinetic sonic
very narrow peak. However, the actual spectral amplitude (ESA). These two phenomena are both
peak has considerable line width attributable to the known as electro-acoustic effects. The magnitude of
Brownian motion of the particles and a distribution these effects is directly proportional to the electro-
of electrophoretic mobilities. The separation of these phoretic mobility of the particles. The mobility determ-
two sources of spectral-peak broadening is critical ined by either electro-acoustic effect is the dynamic or
because, without such a separation, a measurement AC mobility of the particle.
of electrophoretic mobility at a single angle provides The dynamic mobility which is determined by the
information only about the peak position (Doppler electro-acoustic effects differs from the low-frequency
shift). The shape of the peak (Doppler broadening), or DC mobility that is determined by electrophoresis
being attributable to either of the two effects, cannot
be interpreted with a single-angle measurement. Simul-
taneous multiple-angle measurements allow peak Photo- Cathode Variable Collimating Beam
diodes attenuator lens shaper
Water–Zeta

shapes to be analyzed to give accurate measurement Main

beam
of the electrophoretic mobility and the subsequent Laser
determination of zeta potential according to the
Helmholtz–Smoluchowski equation [Eq. (1)].
Beam Focusing Frequency Beam Focusing
The DELSA 440 (Coulter Scientific Instruments) is dump
Anode lens shifter splitter lens
a laser-scattering particle electrophoretic analyzer that
measures the electrophoretic mobility distribution and Fig. 4 A schematic diagram of the DELSA 440 system.
 4120 Zeta Potential

because of the effects of particle inertia which cause

particle velocities in an alternate field to become out Synthesizer
of phase with the applied field. The higher the
frequency, the greater is the phase lag between the par-
ticle and the applied field. Dynamic mobility (Ud) is a
function of frequency, particle size, particle density,
and particle zeta potential. These inertial effects arise Gated Measurement Signal 80386
in both ESA and CVP measurements. In the case of Amplifier Cell Processing Computer
spherical particles with thin double layers and low zeta
potential, the equation below can be applied: Fig. 5 An AcoustoSizer signal processing block diagram.

Ud ¼ ðez=ZÞGðaÞ ð3Þ
converted by a pressure transducer in the cell to an
electrical signal, which then passes to the signal proces-
where G(a) is the inertial term. The formula for sing electronics. The electronics extract the amplitude
dynamic mobility is identical to the well-known and the phase of the ESA signal, storing them in the
Helmholtz–Smoluchowski equation [Eq. (1)] for DC computer housed in the AcoustoSizer.
electrophoretic mobility except for the G(a) term. A schematic diagram of the cell is shown in Fig. 6.
A linear relationship exists between the ESA or The dispersion is located in the region between the pair
CVP amplitude and the volume fraction of the sus- of electrodes. The pulse from the gated amplifier is
pended particles. At relatively high-volume fractions, applied across these electrodes, generating the ESA
hydrodynamic and electric double-layer interactions sound waves. Voltage pulses rather than continuous
lead to a non-linear dependence of these two effects sinusoids are applied to avoid interference by the
on volume fraction. Generally, non-linear behavior sound wave emanating from the electrodes. The use
can be expected when the electric double-layer thick- of pulsed signals also avoids electrical heating of the
ness is comparable to the interparticle spacing. In most suspension and the complications of multiple reflec-
aqueous systems, where the electric double layer is tions of the waves at the electrodes and at the ends
thin relative to the particle radius, the electro-acoustic of the rods. The voltage pulse produced by the sound
signal will remain linear with respect to volume frac- wave in the transducer on the right passes into the
tion up to 10% by volume. At volume-fractions that signal-processing electronics, which measures the
are even higher, particle-particle interactions lead to amplitude and phase of the sinusoidal component of
a reduction in the dynamic mobility. the pulse. These data are passed to the computer where
The commercial systems for zeta-potential determi- it is stored for subsequent processing.
nation using these electro-acoustic effects are typified Since the ESA signal depends on particle motion
by the AcoustoSizer 8000 (Matek Applied Sciences) and also on the acoustic property of the dispersion,
consisting of five main components (Fig. 5). The the determination of particle motion from the ESA sig-
synthesizer produces a continuous sinusoidal voltage nal requires knowledge of the acoustic impedance of
which feeds into the gated amplifier. This creates a the dispersion. This parameter is measured using sen-
sinusoidal voltage pulse across the cell which contains sors associated with the rod on the left immediately
the dispersion. The resulting ESA sound waves are after each ESA measurement. The signals from these

Signal From
Gated Amplifier
Acoustic Impedance Pressure
Sensing Element Transducer

Colloid

Acoustic Impedance
Measurement Rod ESA Measurement
Water–Zeta

Acoustic Delay Rod

Electrodes

To Signal To Signal
Processing Processing

Fig. 6 A schematic diagram of AcoustoSizer measurement cell.

Zeta Potential 4121

two rods are stored in the computer and are subse- be positive (repulsion) at both below and above this
quently converted to a particle-velocity spectrum, and concentration.
from that the zeta potential and size information is The attractive-force component between particles
determined. The AcoustoPhor 8000 (PenKem, Inc.) is in a colloidal system is developed by summation of
another example of a commercial system which deter- the London dispersion forces between all atom pairs
mines zeta potential and the particle size of dispersions in the particles. Neglecting retardation effects, the
using these electro-acoustic effects. expression for the attractive energy VA between two
particles of radius a at a distance of separation H0,
for a  H0, is
APPLICATIONS
VA ¼ A; a12H0 ð5Þ
The practical significance of zeta-potential measure-
ment lies in the fact that strong empirical correlation where A is the effective Hamaker constant describing
exists between the measured zeta potential of the sys- the attraction between the particles and the dispersion
tem and the properties of the system which are the medium. It is apparent that VA decreases as an inverse
manifestation of the electrostatic interfacial phenom- power of the distance between the particles.
enon. Since the measurement of zeta potential can be According to DLVO theory, the total energy of
conveniently performed, it becomes an ideal parameter interaction between colloidal particles is given by the
for use in routine testing. Zeta-potential control has sum of the attraction (VA) and repulsion (VR) energies:
been successfully applied to various technical fields
involving colloidal and non-colloidal systems. VT ¼ VR þ VA ð6Þ

Fig. 7A shows the total interaction energy curve for a

Colloid Stability[3,8,9] colloidal system. At very close distances between the
particles, the Born repulsion between adjoining elec-
The physical stability of a colloidal system is determ- tron clouds predominates, and the net interaction is
ined by the balance between the repulsive and attract- repulsion. However, at slightly greater distances, the
ive forces which is described quantitatively by the van der Waals attraction predominates over the elec-
Deryaguin–Landau–Verwey–Overbeek (DLVO) theory. trostatic repulsion, and the net interaction is strong
The electrostatic repulsive force is dependent on the attraction as shown by the deep potential energy mini-
degree of double-layer overlap and the attractive force mum VP. At even greater distances, the net interaction
is provided by the van der Waals interaction; the magni- is moderate attraction (a shallow secondary minimum
tude of both are a function of the separation between the VS) because with increasing distance between two par-
particles. It has long been realized that the zeta potential ticles, electrostatic repulsion weakens more rapidly
is a good indicator of the magnitude of the repulsive than the van der Waals attraction energy. At inter-
interaction between colloidal particles. Measurement of mediate distances, the electrostatic repulsion predomi-
zeta potential has therefore been commonly used to nates and the net interaction becomes repulsion, with
assess the stability of colloidal systems. an energy barrier, Vm.
Electrostatic repulsive energy, VR, at a given inter- The attraction energy arising from van der Waals
particle distance is the work which must be performed forces does not vary significantly with changes in the
to bring the particles to a specific point. Using the surface potential and electric double-layer properties
Debye–Huckel low-potential approximation and of the colloidal particles. By contrast, changes in these
assuming equal spheres, VR can be described by two parameters have significant impact on the repul-
sion energy component of the total interaction energy.
VR ¼ 2peacd 2 lnð1 þ exp½kHÞ ð4Þ Therefore, at greater distances, a further increase in
electrostatic repulsion will result in a total repulsive
where a is the particle radius, H is the distance of sep- net interaction curve without the secondary minimum
aration between the two particles, and k is the Debye (Fig. 7B). However, as the repulsive component of
parameter. The magnitude of VR decreases in an the interaction energy diminishes (i.e., the electrolyte
Water–Zeta

approximately exponential fashion with increasing H concentration increases), the interaction curve becomes
and that its range decreases by increasing k (i.e., by totally attractive and rapid coagulation (controlled by
increasing electrolyte concentration and/or counter- particle-diffusion rate) of the colloidal particles takes
ion charge number). VR can be also influenced by other place (Fig. 7C).
specific factors. Counter-ion adsorption in the Stern An empirical relationship was developed between
layer may cause a reversal of charge, so that VR will the zeta potential and the coagulation behavior of a
be zero at the reversal-of-charge concentration and will variety of systems. Deryaguin shows that a rapid
 4122 Zeta Potential

A B C

VR
Potential Energy

Vm
H

VT

VS
VA

VP

Fig. 7 Variation of the attraction and repulsion energies and the total energy of interaction between two colloidal particles with
interparticle distance.

decrease in stability can be expected when the follow- the size of their dispersed phase. Therefore, in general,
ing conditions are met.[3] the theoretical and empirical relationships between
zeta potential and colloid stability are applicable to
4pex2 =kA ð7Þ the physical stability of these pharmaceutical systems.
However, the practical aspects of applying zeta poten-
For a given system, A and e are fixed; therefore, tial to these systems may vary because of differences
varying the electrolyte (in type and concentration) in the chemical composition and physical state of their
should produce instability at a particular value of dispersed and continuous phases. In addition, the prac-
x2/k. The validity of using zeta potential in describing tical significance of zeta-potential measurement may be
the coagulation process should come as no surprise different for products which differ in their clinical
because it characterizes the potential of the diffuse part requirements such as route of administration and
of the electric double layer which is involved in double- consequent in vivo disposition.
layer overlap and which plays an important role in the
coagulation process. When the zeta potential is suffi-
ciently high to produce a potential barrier opposing Emulsions
coagulation, the rate of coagulation is slowed by a
factor W, called the stability ratio, defined as the ratio Emulsions have been widely used as vehicles for oral,
of the total number of particle collisions to the number topical, and parenteral delivery of medications.
of collisions forming permanent doublets. Wiese and Although the product attributes of an emulsion dosage
Healy reported an excellent correlation between the form are dependent on the route of administration, a
theoretical values for W (calculated from the zeta common concern is the physical stability of the system,
potential) and the experimental values for TiO2 and in particular the coalescence of its dispersed phase
Al2O3 sols as a function of pH (Fig. 8) and electrolyte and the consequent alteration in its particle-size
(KNO3) concentration.[10] Rapid coagulation, whether distribution and phase separation. The stabilization
induced by pH or KNO3 concentration changes, mechanism(s) for an emulsion is mainly dependent
occurs at a value of jzj  14  4 mV, corresponding on the chemical composition of the surfactant used.
Water–Zeta

to z2/k ¼ 6
104 (mV)2 cm. Electrostatic stabilization as described by DLVO

theory plays an important role in emulsions (O/W)
containing ionic surfactants.[11] For O/W emulsions
Pharmaceutical Systems with low electrolyte content in the aqueous phase,
a zeta potential of 30 mV is found to be sufficient to
Most of the pharmaceutical dispersions can be classi- establish an energy maximum (energy barrier) to
fied as colloidal systems, although this depends upon ensure emulsion stability.[11] For emulsions containing
Zeta Potential 4123

50 the surface potential are likely to have an impact on

Theoretical overall emulsion stability. The physical instability of
Experimental an injectable emulsion as manifested by an increase
in particle size (>5 mm) will result in serious clinical
10–4M KNO3 consequences such as formation of pulmonary
20
emboli.[15] Therefore, the extent of electrostatic stabili-
zation of an injectable emulsion as measured by its zeta
potential has been an important parameter in the char-
10 acterizing and monitoring of its physical stability.
A relative wealth of information relating to the
Stability Ratio

application of zeta potential to injectable emulsions

5 has been documented with respect to the use of total
nutrient admixtures (TNA).[16] Total nutrient admix-
tures are prepared by mixing the lipid emulsion with
other components (i.e., dextrose, amino acids, and elec-
trolytes) in a single container prior to administration.
2 Depending on composition, the mixtures vary widely
in their stability and may show clinically unacceptable
coalescence after different periods of storage time.
1 When the electrostatic stabilization of the emulsion
is considered, the electrolytes (monovalent and
TiO2 Al2O3
divalent) added to the mixture are the major destabiliz-
ing species. The zeta potential of the emulsion particles
5 6 7 8 9 10 is a function of the concentration and type of electro-
pH
lytes present. Two types of emulsion particle-electro-
Fig. 8 Comparison of theoretical and experimental stability lyte (ions) interaction are proposed: non-specific and
ratios for TiO2 (0.05 g/L) and Al2O3 (0.15 g/L) colloids in specific adsorption.[16] In non-specific adsorption the
104 M KNO3. ions are bound to the emulsion particle only by electri-
cal double-layer interactions with the charged surface.
As the electrolyte concentration is increased, the zeta
non-ionic surfactants (polymers), the principal stabili- potential asymptotes to zero. As the electrostatic
zation mechanism is the repulsion between the repulsion decreases, a point can be found where the
adsorbed polymer chains on the surface of the oil attractive van der Waals force is equal to the repulsive
globules called steric stabilization.[11] In such a case, electrostatic force and flocculation of the emulsion
zeta-potential measurement may have limited values occurs (Fig. 9A). This point is called the critical floccu-
with respect to emulsion stability. lation concentration (CFC).
In the past, injectable emulsions were administered DLVO theory predicts that the CFC should vary
intravenously to provide patients with vegetable oils inversely with the sixth power of the ion charge. The
as a major energy source in nutrition therapy.[12] In application of CFC alone fails to correlate the TNA
recent years, injectable emulsions have been found to stability with electrolytes in the mixtures. This is
be very effective vehicles for the parenteral delivery because divalent ions (calcium and magnesium)
of water-insoluble drugs.[13] Because of toxicological adsorbed specifically on the emulsion particles through
considerations, surfactants which are used in the for- their complexation with the negatively charged acidic
mulation of injectable emulsions have been largely phospholipids. Fig. 9B demonstrates the relationship
limited to mixtures of phospholipids which are either between the concentration of a specifically adsorbed
derived from egg (egg yolk lecithin) or soybean (soy- ion and the zeta potential and flocculation rate of an
bean lecithin). The majority of the phospholipids injectable emulsion. The increase in electrolyte concen-
(80–90%) are phosphatidylcholine (PC) and phosphati- tration causes the particle charge to pass through zero;
dylethanolamine (PE), which are uncharged at physio- this is referred to as the point of zero charge (PZC).
Water–Zeta

logical pH. But the presence of small quantities (2–5%) At this point the flocculation rate is at maximum; a
of acidic components, largely phosphatidylserin (PS) further increase in electrolyte concentration causes a
and phosphatidylglycerol (PG), which are ionized at surface-charge reversal of the particles and increases
pH 7, confers a surface charge of approximately 40 in zeta potential (opposite sign) as well as a decrease
to 50 mV on the emulsion particles.[14] Therefore, of the flocculation rate from its maximum value.
the stability of these emulsions is mainly attributable The effect of pH on the stability of an injectable
to electrostatic stabilization. Any factors which alter emulsion was also followed by measuring its zeta
 4124 Zeta Potential

A B
Zeta Potential Zeta Potential

+ Electrolyte Concn. +
0 0
– –
Electrolyte Concn.

Flocculation Rate Flocculation Rate

C.F.C
C.F.C

Electrolyte Concn. Electrolyte Concn.

Fig. 9 Zeta potential and flocculation rate of a parenteral emulsion in the presence of a non-specifically adsorbing electrolyte (A)
and a specifically adsorbing electrolyte (B).

potential (Fig. 10). At pH 7 the ionization of PS and using DLVO-based methods in the measurement of the
PG imparts a surface negative charge (30 to zeta potential of the system.[19]
50 mV) to the emulsion particles. As the pH is
reduced, this ionization is suppressed until the charge Suspensions
is zero at a pH of 3.2. Further decreases in pH cause
the protonation of the phospholipid and a positive Pharmaceutical suspensions are dispersions of solid
surface charge (positive zeta potential). Significant particles in a suspending medium or vehicle (usually
progress has been made in relating the electrokinetic aqueous in nature). When the suspended solids are less
properties of phospholipid-stabilized emulsions to their than 1 mm, the system is referred to as a colloidal sus-
instability when influenced by other TNA ingredi- pension. When the particle sizes are greater than about
ents.[17,18] Washington and his coworkers have attempted 1 mm, the system is called a coarse suspension. The
to develop a model to predict emulsion stability in TNAs practical upper limit for particles in a coarse suspen-
sion is approximately 50–75 mm. Depending on the
affinity or interaction between the dispersed phase
and the dispersion medium, a colloidal dispersion can
Zeta/mV be classified as lyophilic (hydrophilic) or lyophobic
20 (hydrophobic).[20]
The dispersion of hydrophilic particulate solids in
water occurs spontaneously and gives rise to a thermo-
0 dynamically stable system such as colloidal silicone
dioxide or microcrystalline cellulose. Hydrophobic
solids are not easily wetted and are not dispersed
–30 spontaneously in water as sulfur, clays, and most
non-polar organic compounds. The van der Waals
attractive forces between particles cause them to
aggregate, since the solvation forces which promote
–40
dispersal in water are weak. Therefore, aqueous disper-
Water–Zeta

sions of hydrophobic solids can only be stabilized kine-

tically to resist flocculation and coagulation, which can
–60
be defined by referring to the total interaction energy
2 4 6 8 10
curve (Fig. 7). Flocculation indicates the association
pH
of particles in the secondary minimum and coagulation
Fig. 10 Zeta potential of a parenteral emulsion as a function is the aggregation of particles in the primary minimum.
of pH. Since the attractive forces between particles in a
Zeta Potential 4125

flocculated system are relatively week, they can be

100 Caking zone Noncaking zone Caking zone
redispersed and the process is reversible. In contrast, + + –
+ + – + – – VeVa
coagulation is irreversible as attractive forces operating +
+
+ +
+
– –
–
–
Ratio
between particles in the primary minimum are difficult

Apparent zeta potential

to overcome. 0.06
Ve / V a Curve
When electrostatic repulsion is predominant
between particles in a suspension, it is called a defloc- + Zeta potential
0.03
curve
culated system. For particles with a diameter of
2–5 mm, Brownian movement counteracts sedimen-
tation to a measurable extent at room temperature 0 0
by keeping the dispersed particles in random motion.
This results in a system which is physically stable with –
respect to sedimentation and flocculation.[20] However,
Caked Not caked Caked
in a deflocculated suspension consisting of larger parti- 50
cles (coarse), gravitational force counteracts Brownian Concentration KHe POi
movement and sedimentation occurs.[21] At the bottom
of the container, a compact sediment with strong Fig. 11 A caking diagram showing the flocculation of a bis-
attractive forces between the particles is formed and muth subnitrate suspension by means of the flocculating
it cannot be easily redispersed. This phenomenon is agent, potassium monobasic phosphate.
referred to as caking. The prevention of caking has
been one of the main objectives in preparing stable
coarse suspensions.[21,22] Although flocculation is then increases in a negative direction. The suspension
referred to as a physical instability of colloids, floccu- remains flocculated until the zeta potential becomes
lated suspensions are pharmaceutically acceptable sufficiently negative to effect deflocculation of the sys-
since they are non-caking. In a flocculated suspension, tem, which is shown by a decrease in the sedimentation
particles attract each other (at the secondary mini- volume and an increase in caking tendency.
mum) loosely to form flocs which tend to settle Zeta potential has also found an application in
together, creating a distinct boundary between the relating the surface-charge characteristics of a suspen-
sediment and the supernatant. Particles in flocs are sion to properties other than physical stability. Heyd
thereby easily redispersed after settling. and Dhabhar improved the fluidity of concentrated
Controlled flocculation is a process by which floccu- antacid suspensions by adding a colloidal polyelectro-
lation of particles in a suspension is purposely pro- lyte (carrageenan sodium) which is called a fluidizing
duced by adding flocculating agents. One of the most agent.[25] As the concentration of the fluidizing agent
effective methods of achieving controlled flocculation increases, the zeta potential of the suspension changes
is to reduce the electrostatic repulsive forces between from positive to negative, accompanied by a reduction
particles.[21,22] Electrolytes have been shown to be in viscosity of the suspension. The mechanism for this
effective flocculating agents. Zeta-potential measure- viscosity-thinning phenomenon is believed to be the
ment has been a valuable tool in monitoring and selective adsorption of the negatively charged fluidizing
evaluating the influence of various electrolytes on the agent onto the positively charged antacid particles. This
flocculation phenomena in coarse suspensions.[23] Sedi- imparts an electronegative charge (as measured by the
mentation volume F, which is the ratio of the volume zeta potential) onto the antacid particles, decreasing
of sediment to the original volume of the suspension, the demand for water.
is a useful parameter in measuring the extent of floccu- In the literature, the reported use of zeta potential
lation of the suspension. An excellent correlation measurement for non-aqueous suspensions is relatively
between the zeta potential, sedimentation volume, and infrequent because non-aqueous suspensions only rep-
caking was found for a series of bismuth subnitrate resent a small percentage of all medicated suspensions.
suspensions containing various concentrations of Su and others evaluated the flocculation–deflocculation
potassium phosphate monobasic (Fig. 11).[24] The behavior of cefazolin sodium in non-aqueous media
addition of potassium phosphate monobasic causes a and the effect of surfactants as measured by zeta
Water–Zeta

decrease in the zeta potential of the suspension because potential along with sedimentation and porosity mea-
of partial neutralization of the positive surface charge surements.[26] A significant difference in zeta potential
on the bismuth subnitrate particles. With the decrease was observed when the particles were dispersed in
in the zeta potential (positive), the suspension becomes peanut oil and ethyl oleate. The addition of lecithin
flocculated and non-caking, accompanied by an increase reduced the zeta potential of cefazolin sodium, result-
in the sedimentation volume. The continued addition of ing in a deflocculated state accompanied by a decrease
electrolyte causes the zeta potential to fall to zero and in sedimentation volume. The effect of surfactant
 4126 Zeta Potential

excipient on the zeta potential of salbutamol in tri- charged liposomes, their surface potential is a critical
chlorofluoroethane (P113) for metered-dose inhalation factor affecting their physical stability and in vivo
was determined.[27] Although the zeta potential of the performance. Instead of their surface potential, zeta-
particles is decreased (more negative) by the addition potential measurement is frequently used in the investi-
of oleic acid, the overall zeta potential is too low to gation of the behavior of liposomes as influenced by
be effective in providing the system with adequate elec- their surface charge. Liposomes in aqueous dispersions
trostatic stabilization, particularly in media with a low have a tendency to aggregate and subsequently fuse on
dielectric constant (organic solvents). storage. This physical instability potentially limits their
Microparticulate drug-delivery systems, including application as dosage forms.
microspheres and nanoparticles, are suspensions when Crommelin incorporated charge-inducing compo-
they are administrated in a liquid medium. Micro- nents in liposome bilayers to produce electrostatic
spheres are solid particles containing dispersed drug repulsion between liposomes, thus increasing the shelf
in either solution or microcrystalline form and range life of the dispersion.[35] He investigated the effect of
in size from 1 to 1000 mm. Although most microspheres including stearylamine or phosphatidylserin on the zeta
are outside the conventional colloidal size range, in potential of phosphatidylcholine-cholesterol–containing
pharmaceutical literature microspheres with sizes up liposomes. A correlation between the aggregation stabil-
to approximately 15 mm are still considered as colloidal ity of negatively charged liposomes and the increments
delivery systems. Nanoparticles are similar to micro- in the surface-charge density at both low and high ionic
spheres but have particle sizes in the range of 0.01– strength was established, whereas the positive liposome
1 mm; therefore, they form colloidal suspensions when dispersions were found to be unstable.
dispersed in a liquid medium.[28] Zeta-potential measure- Carrion and his coworkers investigated the effect of
ment is an important surface-characterization technique incorporating phosphatidic acid on the zeta potential
which provides information regarding the surface charge and aggregation of large unilamellar vesicle liposomes
of these microparticulate systems. The effect of surface of phosphatidylcholine in the presence of neutral elec-
charge (zeta potential) on the physical stability of trolytes.[36] Their results show that increasing concen-
these systems (i.e., aggregation) can be predicted using trations of phosphatidic acid in lipid bilayers resulted
the DLVO theory, although steric stabilization plays in high zeta potential and enhanced physical stability
an important role in systems with surface-adsorbed of the liposomes. The destabilizing effect of divalent
macromolecules. cations (i.e., calcium ions) on charged liposomes has
When the microparticles are administered intra- been well documented and is attributable to the
venously, surface change is an important parameter reduced surface potential of the liposomes as the result
affecting the interaction between the microparticles of cation binding.[37–39]
and other biological components of the body and their The effect of calcium ions on the aggregation behav-
subsequent in vivo distribution and disposition.[29] iors of neutral liposomes composed of egg phos-
Tabata and Ikada reported that phagocytosis of poly- phatidylcholine and dipalmitoylphosphatidylcholine
mer microspeheres by macrophages is enhanced as the was investigated by Mosharraf and coworkers.[40] Both
absolute value of zeta potential increases for both the systems displayed a high negative zeta potential in
negatively charged and the positively charged surfaces, deionized water, probably the result of adsorption of
while the lowest phagocytosis is shown for the surface hydroxyl ions. A reasonable correlation was found
with a zeta potential of zero.[30] The uptake of charged in fboth cases between particle-size increase and zeta
serum components by nanoparticles (opsonization) in potential decrease of the system, suggesting that the
blood is considered to be the first step leading to mechanism by which the aggregation takes place is
phagocytosis by liver and spleen macrophages. Muller related to the surface-charge characteristics. Although
and his coworkers determined the increase in zeta it has long been established that liposomes composed
potential of non-particles in serum as a measure of of neutral phospholipids acquire negative surface
the extent of opsonization and used this as selection charge via anion adsorption, Makino and his group
criteria for i.v. drug carriers avoiding liver/spleen in Japan found that neutral liposomes exhibit negative
uptake.[31] zeta potential in buffer solution containing no chloride
ion.[41] They also reported that changes in the ionic
Water–Zeta

Liposomes strength and temperature causes the zeta potential of

neutral liposomes to reverse sign. They proposed that
Liposomes are phospholipid-based vesicles which have the reversal of zeta potential is triggered by changes
been studied as delivery systems to target drugs to spe- in the direction of the dipole connecting the negative
cific sites in the body.[32–34] The surface of a liposome charge of the phosphatidyl group and the positive
can be neutral or negatively or positively charged, charge of the choline group consisting the head group
depending on the composition of lipids used. For of a lipid molecule.
Zeta Potential 4127

The study of the effect of drug-loading on the zeta it allows the quantitation of the electrostatic repulsion
potential of liposomes provides information regarding between particles, which is one of the most important
the drug-liposome interaction. Lawrence and others forces governing the behavior and physical stability
reported that the loading of polymyxin B, a polycatio- of colloidal systems. In recent years, the concept
nic antibiotic in negatively charged liposomes, results of zeta potential has been applied to areas beyond
in the decrease of zeta potential, indicating electrostatic classical colloidal sciences and industrial processes.
attraction between the positive drug and the negative Pharmaceutical sciences are the new fields to which
lipid bilayer.[42] That the highest drug-loading is found the application of zeta potential has been extended.
in negatively charged liposomes in comparison to The expanding role of zeta potential in these fields is
positively charged and neutral liposomes further sup- attributable to the advance in modern instrumentation
ports this electrostatic drug-liposome interaction. of zeta potential measurement, the rapid development
Beschiaschvili and Seelig investigated the binding of of colloidal drug-delivery systems, and the emphasis
a cyclic somatostatin analogue, a positively charged on interdisciplinary basic research. While good corre-
peptide, to negatively charged mutilamellar lipo- lation has been demonstrated for zeta potential and
somes.[43] From the binding isotherm and the zeta- in vitro properties of colloidal drug-carrier systems,
potential measurement, they were able to describe a the understanding of the impact of zeta potential in
partition equilibrium between the peptide and the organ distribution and in vivo clearance of these sys-
negatively charged membrane with a surface-partition tems has been the focus of additional research efforts.
constant. They concluded that most of the peptide
molecules are embedded in the headgroup region with
little penetration into the lipid core.
The measurement of the zeta potential of liposomes ARTICLE OF FURTHER INTEREST
provides valuable information relating to their in vivo
performance because, in addition to size, the surface Microsphere Technology and Applications, p. 2328.
charge of liposomes is an important determinant of
their clearance from the general circulation and their
tissue disposition after parenteral administration. REFERENCES
Large multilamellar liposomes are cleared from the cir-
culation much more rapidly than small unilamellar 1. Shaw, D.J. Introduction to Colloid and Surface Chemistry,
liposomes. Among small unilamellar liposomes, how- 4th Ed.; Butterworth Heinemann Ltd.: Oxford, 1992.
2. Goetz, P.J.; Penniman, J.G. A new technique for microelec-
ever, those with a negative surface charge are cleared trophoretic measurements. Jr. Am. Lab. Oct. 1976.
rapidly, but positively charged or uncharged liposomes 3. Hunter, R.J. Zeta Potential in Colloid Science, Principles
remain in the circulation for longer periods.[44] and Applications; Academic Press: London, 1981.
4. Uzgiris, E.E. Progress in Surface Science; Pergamon Press:
Rahman and his coworkers compared the pharmaco- New York, 1981; X.
logical and therapeutic effects of adriamycin entrapped 5. Oja, T.; Bott, S.; Sugrue, S. Doppler electrophoretic light
in positively charged and negatively charged liposomes scattering analysis using the coulter DELSA 440. Coulter
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in rats.[45] They found that less cardiotoxicity was 6. O’Brien, R.W. J. Fluid Mech. 1988, 190, 71–86.
associated with the drug entrapped in the positively 7. Babchin, A.J.; Chow, R.S.; Sawatzky, R.P. Electrokinetic
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Interface Sci. 1989, 111–151, 30
uptake by cardiac tissue. When liposomes are 8. Hiemenz, P.C. Principles of Colloid and Surface Chemis-
employed to target drugs to lymph nodes, their surface try, 2nd Ed.; Marcel Dekker, Inc.: New York, 1986.
charge has been shown to affect their lymphatic uptake 9. Hirtzel, C.S.; Rajagopalan, R. Colloidal Phenomena,
Advanced Topics; Noyes Publications: New Jersey, 1985.
from subcutaneous and intraperitonial injection 10. Wiese, G.R.; Healy, T.W. Coagulation and electrokinetic
sites.[46] Patel showed that negatively charged lipo- behavior of TiO2 and Al2O3 colloidal dispersions. J. Colloid
somes yield the highest localization in the lymph nodes, Interface Sci. 1975, 51, 427–433.
11. Friberg, S.E.; Goldsmith, L.B.; Hilton, M.L. Theory of
followed by positively charged and neutral ones. The Emulsion in Pharmaceutical Dosage Forms: Dispersion
drainage of negatively charged liposomes has also Systems; Lieberman, H.A., Reiger, M.M., Banker, G.S.,
shown to be faster than that of positive liposomes after Eds.; Marcel Dekker, Inc.: New York, 1988; I.
12. Wretlind, A.J. Development of fat emulsions. Parenter.
intraperitoneal administration.[47] Enteral. Nutr. 1981, 5, 230–235.
Water–Zeta

13. Davis, S.S.; Washington, C.; West, P.; Illum, L.; Liversidge,
G.; Sternson, L.; Kirsh, R. Ann. N.Y. Acad. Sci. 1987, 507,
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CONCLUSIONS 14. Washington, C.; Chawla, A.; Christy, N.; Davis, S.S. The
electrokinetic properties of phospholipid-stabilized fat
Zeta-potential measurement has long been recognized emulsions. Int. J. Pharm. 1989, 54, 191–197.
15. Burnham, W.R.; Hansrani, P.K.; Knott, C.E.; Cook, J.A.;
as an excellent tool for characterizing colloidal sys- Davis, S.S. Stability of a fat emulsion based intravenous
tems. Its practical significance stems from the fact that feeding mixture. Int. J. Pharm. 1983, 13, 9–22.
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16. Washington, C. The stability of intravenous fat emulsions 33. Gregoriadis, G.; Florence, A.T. Liposomes in drug delivery.
in total parenteral nutrition mixtures. Int. J. Pharm. 1990, Drugs 1993, 45, 15–28.
66, 1–21. 34. Barenholz, Y.; Crommelin, D.J.A. Liposomes as pharma-
17. Washington, C.; Athersuch, A.; Kynoch, D.J. The electro- ceutical dosage forms. In Encyclopedia of Pharmaceutical
kinetic properties of phospholipid stabilized fat emulsions. Technology, 1st Ed.; Swarbrick, J., Boylan, J.C., Eds.; Mar-
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64, 217–222. 35. Crommelin, A.J.A. Influence of lipid composition and ionic
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lized fat emulsions. V. The effect of amino acids on emul- 36. Carrion, F.J.; De La Maza, A.; Parra, J.L. The influence of
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19. Washington, C.; Ferguson, J.A.; Irwin, S.E. Computational liposomes. J. Colloid Interface Sci. 1994, 164, 78–87.
prediction of the stability of lipid emulsions in total nutrient 37. Düzgünes, N.; Sur, S.; Wilschut, J.; Bentz, J.; Newton, C.;
admixtures. J. Pharm. Sci. 1993, 82, 808–812. Portis, A.; Papahadjopoulos, D. Calcium- and magnesium-
20. Zograft, G.; Schott, H.; Swarbrick, J. Dispersion systems. induced fusion of mixed phosphatidylserine/phosphatidyl-
In Remington’s Pharmaceutical Sciences, 18th Ed.; Mack choline vesicles: effect of ion binding. J. Membr. Biol.
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21. Hiestand, E.N. Theory of coarse suspension formulation. 38. Hope, M.J.; Walker, D.C.; Cullis, P.R. Ca2þ and pH
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22. Falkiewicz, M.J. Theory of suspensions in theory of emul- phosphatidylethanolamine and negatively charged phos-
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Lieberman, H.A., Reiger, M.M., Banker, G.S., Eds.; Marcel Commun. 1983, 110, 15–22.
Dekker, Inc.: New York, 1988; I. 39. Minami, H.; Inque, T.; Shimozawa, R. Aggregation kinetics
23. Nash, R.A.; Haeger, B.E. Zeta potential in the development of dimyristoylphosphatidylglycerol vesicles induced by
of pharmaceutical suspension. J. Pharm. Sci. 1966, 55, 829–837. divalent cations. J. Colloid Interface Sci. 1993, 158, 460–465.
24. Martin, A.; Swarbrick, J. American Pharmacy, 6th Ed.; 40. Mosharraf, M.; Taylor, K.M.G.; Craig, D.Q.M. Effect of
Lippincott: Philadelphia, 1966. calcium ions on the surface charge and aggregation of phos-
25. Heyd, A.; Dhabhar, D.J. Enhancing fluidity of concentrated phadylcholine liposomes. J. Drug Target. 1995, 2, 541–545.
antacid suspensions. J. Pharm. Sci. 1975, 64, 1697–1699. 41. Makino, K.; Yamada, T.; Kimura, M.; Oka, T.; Ohshima, H.
26. Su, K.S.E.; Quay, F.; Campanale, K.M.; Stucky, J.F. Temperature and ionic strength-induced conformational
Nonaqueous cephalosporin suspension for parenteral changes in the lipid head group region of liposomes as
administration: cefazolin sodium. J. Pharm. Sci. 1984, 73, suggested by zeta-potential data. Biophy. Chem. 1991, 41,
1601–1602. 175–183.
27. Clarke, J.G.; Wicks, S.R.; Farr, S.J. Surfactant mediated 42. Lawrence, S.M.; Alpar, H.O.; McAllister, S.M.; Brown,
effects in pressurized metered dose inhalers formulated as M.R.W. Liposomal (MLV) polymyxin B: physicochemical
suspensions. I. Drug/surfactant interactions in a model characterization and effect of surface charge on drug associ-
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Marcel Dekker, Inc.: New York, 1994; 10. tostatin analogue. Biochemistry 1990, 29, 10,995–11,000.
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30. Tabata, Y.; Ikada, Y. Effect of the size and surface charge Woolley, P.; Schein, P.S. Liposomal protection of adriamy-
of polymer microspheres on their phagocytosis by macro- cin-induced cardiotoxicity in mice. Cancer Research 1980,
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Eds.; Marcel Dekker, Inc.: New York, 1989; 2. 1984, 801, 76–86.
Water–Zeta
Index

AAG. See Alpha acid glycoprotein. [Absorption] Absorption spectroscopy, in chemical

AAG-bound erythromycin, 3037 oral, 22 component quantitation, 3471
AB block copolymer micelles, 3588 parenteral and enteral, 19 Absorptive cell basolateral membrane, 2715
hydrophobic drugs, 3588 pharmacokinetic characterization, 19 Acacia
Abbreviated new drug application (ANDA), process of, 214 gum, 994–995
1787, 3188, 3931 of protein molecules, 2731 suspending agents, 1624
bioequivalence for, 1892 pulmonary, 2733 Accelerators, sulfur-cure, 1467
and stability information systems, 733 rectal, 22 Acceptable daily intake (ADI), 1588
Abciximab (ReoProÕ), 1569 sublingual, 21 Accreditation and certification standards
AbelcetÕ, 3362 vaginal, 22 for developing and programming, 2167
drug product, 1643 Absorption, distribution, metabolism, for testing and calibration laboratories,
Abietic acid excretion (ADME) pharmacokinetic 2167
chromatogram of, 1694 studies, 1785 validation requirements, 2167
molecular structure of, 1694 Absorption enhancers, 30, 2685 Accreditation program, objectives of, 125
Ab initio calculations, 718 alcohols and polyols, 13 AccudepÕ technology, 999
electron correlation effects, 719 amines and amides, 13 AcDiSolÕ, 2010, 3555, 3557, 3566
Gaussian type (GT) functions, 719 in animal models, 2733 compression pressure disintegration,
hydrogenic orbitals, 719 bile salts, 31 3358–3359
linear combination of atomic orbital characteristics of, 13 Acellular hemoglobin solution, infusion of,
(LCAO) method, 719 chemical, 13 372
molecular orbital (MO) theory, 719 coadministration of, 2726 Acellular perussis, 4101
non-parameterized method, 719 cyclodextrins, 14 Acetaminophen, 943, 1385, 2416, 3438
polarization functions, 719 damaging effects of, 2681 active ingredients, 4106
Slater type (ST) functions, 719 enzymatic activities, inhibition of, 2685 atomic force microscopy (AFM)
Abnormal toxicity, test for, 2830 esters, 14 images of, 40
AbraxaneTM, 268, 3361 fatty acids, 13 capsule, 3073
nanoparticle formulation, 2384 functions of, 2685 cocrystals, 618
protein-stabilized, 2384 liposomes, 31 etching pattern of, 40
Absorbance spectra mixed micelles, 31 hepatotoxicity, 319
for dihydrotestosterone, 452 nasal, 2685 isothermal antisolvent crystallization of,
for testosterone, 452 oral, 31 867
Absorbent gauze, in wound dressing, pharmacopeiae liver toxicity by, 2426
1026 mucosal application, 2685 release, 2019
Absorption, 1311 for pulmonary delivery, 2733 solid dispersion of, 779
active transport mechanism of, 2716 sulfoxides, 14 sustained-release tablets, 1235
bases, 3259 surfactants, 31 Acetaminophen tablet
anhydrous types of, 3260 terpenes, 14 extragranular crospovidone in, 3564
classification of, 3259 types of, 2713 manufacture on roller compaction, 2409
buccal, 21 use of, 13, 1256, 2684, 2685 Acetate-shikimate biosynthesis, 237
cell membrane, 19 Absorption filters Acetic acid preparations
clinical factors, 19 bandpass, 3396 antibacterial activity, 2477
definition of, 19 cut-off, 3396 antifungal activity, 2477
degradation, 19 neutral tint, 3396 in external ear, 2478
of drugs, 19, 165 Absorption measurements, 201 Acetohexamide dispersion, by spray–drying
enhancement, mechanism of, 2674 excitation wavelength, 201 process, 775
enteral, 22 flow cells, 201 Acetonitrile, 303
gastrointestinal tract, 22 non-halogenated solvents, 201 Acetylation, 319
acidic environment in, 23 Absorption-promoting adjuvants, variety of, Acetylcholine
liposome for, 31 1307 biosensors, 2030
mathematical aspects of, 1311 Absorption rate constant, 2719, 2819 iontophoresis of, 3822
mechanisms of, 25 Absorption rates, of ibuprofen, 426 pharmacophoric conformations of, 2149
membrane penetration, 19 Absorption spectrophotometry, 449 Acetylcholinesterase, 2030

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.

I1
I2 Index

Acetyl coenzyme A, 231 Acrylic acid (AA), 2025 [Active pharmaceutical ingredient (API)]
catalysis in cytosolic N-acetyltransferases, polymers, 768 non-toxic impurities, 3940
319 synthetic, 1611 objective, 3935
Acetylcysteine, in acetaminophen toxicity Acrylic adhesive, 2927 preapproval inspection, 3935
treatment, 3942 Acrylic alkyl esters, copolymer of, 2927 process optimization, 3935
Acetylethyl tetramethyltetralin, restriction in Acrylic copolymers, lipophilicity of, 2930 process qualification, 3935
cosmetics, 801 Acrylic microspheres, controlled release of, particle morphology, 2340
Acetyl-L-carnitine 2409 particle size distribution, 2340
in Alzheimer’s disease treatment by, 2440 Acrylic oil–gel adhesive, 2928 particle size of, 2339
side effects, 2440 Acrylonitrile rubber, 2275 particle surface properties, 2348
Acetylsulfonamides, water-soluble, 319 Acryloyl-polyethylene glycol–RGDS, 2030 physical characteristics, 2339, 3939–3940
Acetyltributyl citrate (ATBC), 2008 Actinometry, 2863 preformulation studies with, 3930, 3932
Acetyltryptophan, 4002 Actinomyces pyogenes, 2476 propensity of, 632
Achiral molecules, 453 mastitis by, 3958 raw materials in, 2502
Achiral racemic mixture, 451 ActisiteÕ, 903 slurry milling of, 2342
Achlorhydric patients, 2871 Activated clotting enzyme, 3059 solubility of a binary cocrystal of, 621
Acid–base equilibria, neutralization Activated-modulated drug delivery systems stability of, 3306
titrations, 3754 biochemical classification, 1090 stabilizing, 1835
Acid–base indicators, neutralization biochemical-activated, 1090 stage-I product design, 3932
titration, 3755 enzyme-activated, 1090 steps in, 3933
Acid chloride, hydrolytic reactivity of, 2043 chemical classification stage-II process development
Acidic digestive fluids, 2664 hydrolysis-activated, 1090 fractional factorial design for, 3935
Acidic drugs ion-activated, 1090 parameters, 3934
dissolution of, 3181 pH-activated, 1090 pilot laboratory, 3933
salts solubility of, 3180 physical classification process characterization, 3933
sodium salts of, 3177–3178 hydration-activated, 1090 product optimization in, 3933
Acidic endosomes, in hydrolysis, 1331 hydrodynamic pressure-activated, stage-III pilot-production batch stage,
Acidic functional groups, dissociation of, 1090 3935
3390 iontophoresis-activated, 1090 clinical use, 3935
Acidic preservatives, 2226 magnetics-activated, 1090 stage-IV formal process validation
Acidity, 22 mechanical force-activated, 1090 program, 3935
food digestion, 22 osmotic pressure-activated, 1090 change control, 3936
hydrochloric acid secretion, 22 sonophoresis-activated, 1090 dosage form categories, 3936
Acid-labile enzyme, delivery of, 296 vapor pressure-activated, 1090 levels, 3936
Acid-labile hydrazone linkage, 1139 Activator protein 1 (AP-1), 2444 objective, 3935
Acid phosphatase, 1341 Active ingredient, properties of, 1618 SUPAC (scale-up post approval change)
Acid-resistant coating, 941 Active ingredients and excipients, program, 3936
Acid secretion, physiological response of, segregation, 3765 three-batch, 3936
2872 Active pharmaceutical ingredient (API) stereochemical configuration, 3939
Acidulated phosphate-fluoride products, aspect ratio, 2340 surface area, 2339
895 ball milling, 2343 wet (slurry) techniques, 2341
Acne medications, sulfur in, 548 bioavailability of, 2339 Active powder drugs, 2974
Acousto-optical tunable filters, 3375 characteristics, 1835 Active systemic anaphylaxis test (ACA), 123
AcoustoSizer 8000 system compressive force, 2349 Active targeting ligands, 1330
components, 4120 concentrations of, 1834 Acute global oxygen deficit, reduction of, 358
gated amplifier, 4120 crystallization, 2339 Acute hypovolemia, 358
determination degree of reproducibility, 2340 Acute illnesses, consequence of, 2871
particle size dispersions, 4121 dissolution rate, 2339 Acute myelogenous leukemia, 1133
zeta-potential, 4120 dry milling techniques, 2341 Acute myeloid leukemia patients, 1140
signal processing, 4120 and excipients, 3304 Acute myocardial infarction
Acquired immune deficiency syndrome and finished drug products, 3939 antimyosin Fab in, 1151
(AIDS), 281, 2105 impact force, 1837, 2349 in blood coagulation, 273
excipients as process, 1613 ISO quality standards, 3940 diagnostic imaging of, 1151
recognition, 1355 parameters, 3939 radiolabeled antimyosin antibody, 1150
treatment drugs, 2473 primary processes, 3939 Acute myocardial necrosis, 1151
Acridine orange (AO) manufacture of, 2339, 3068, 3938 Acute normovolemic hemodilution, 348
base pair in, 931 for medicinal purposes, 3939 Acute pain, treatment of, 1077
interaction with DNA, milling process Acute pancreatitis, 371
Acromegaly, Sandostatin LARÕ in disadvantages of, 2341 treatment of, 349
treatment of, 178 equipments for, 2340 Acute phase reactant, definition of, 3037
Acrylamide, 2025 equipment design, 2349 Acute renal failure, 2639
emulsion polymerization of, 1183 multimodal distributions, 2340 Acute respiratory distress syndrome, 335
Acrylamide-based composite hydrogel particle density, 2340 Acute toxicity, 2991
dressing, 1030 needle-shaped, 2340 Acute toxicology testing, 117

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I3

AcutrimÕ tablet, 1093 [Adhesive] [Adverse drug reactions (ADRs)]

semipermeable membrane, 1093 island dressings, 1029 in geriatrics, 48
solid-type osmotic pressure-activated drug materials, 413, 2925 in pediatrics, 48
delivery system, 1096 gelatin, 413 case–control designs, 53
Acyl glucuronide metabolite, 3033 hypromellose, 413 causality assessment, 54
Acyl–oxygen fission (Ac), 2040 patches, preparation of, 2670 classification systems of, 46
Acyl transfer reactions, 2040 plasters, 965 concurrent diseases, 48
Acylation, conjugation of drugs, 320 properties and toxicity, 2925 allergic diseases, 49
Acylcarnitines, 1307 Adiabatic microcalorimeters, 3738 hepatic disease, 48
Adams–Bashford multistep equation, Adiabatic saturation temperature, 3884 liver disease, 48, 49
2762 Adipic acid renal disease, 49
Adaptive spline modeling of data (ASMOD), crystals, 42 definition of, 46, 1907
2405 in polyanhydride preparation, 184 detection systems, 52
Add-VantageÕ system, 1008 Adipose tissue, 2634 dosage form and delivery system, 47
Addition reactions, of glutathione Adjuvants dose-related reactions, 47
conjugation aluminum as, 3914 drug–drug interactions, 47
in asymmetric centers, 318 antigens, 3913 drug-related factors, 47
in stereochemistry, 319 in veterinary vaccines, 3914 dosing issues, 47
in stereospecificity, 319 Adobe Acrobat’s free portable document interactions between drugs, 47
Additives, 1630–1632, 1733 format codes, 2560 pharmacodynamic changes for, 47
adriamycin RDFÕ, 1630 ADR. See Adverse drug reactions. pharmacokinetic interactions for, 47
alprostadil, 1630 Adrenergic neurons, 1020 types of drugs, 47
aluminum, 1630 Adsorption in elderly, 1907–1908
BioclateÕ, 1632 of binder solution, 34 formulation effects, 47
calcium gluconate injection, 1630 classification of, 34 gender associated factors, 50
CardiotecÕ, 1630 effects of, 34 genetic factors, 49
Cipro IVÕ, 1630 noncompressible excipients, 34 in health care systems, 55
dexamethasone acetate, 1630 nonspecific, 1306 herbal medicines, 53
EdexÕ, 1630 at solid surface, 34 hypoalbuminemia, 48
epsilon amino caproic acid, 1632 surfactant, 3583 impaired renal function, 49
ergotrate maleate, 1630 Adsorption–desorption isotherms, 43 intramuscular (IM) route
Estradurin InjectionÕ, 1630 Adsorption–desorption process, administration, 47
functions in pharmaceutical formulations, equilibrium of, 4041 intravenous (IV) delivery devices for, 47
1630 Adsorption indicator Karch and Lasagna’s definition of, 46
Gamimune NÕ, 1632 cationic mechanisms for, 49
gamma cyclodextrin, 1630 organic dye, 3753 in medication, 52
Lupron Depot InjectionÕ, 1630 Adsorption on dissolution, effects of, 39 meta-analysis of, 47
meglumine (N-methylglucamine), 1630 Adsorptive stripping voltammetry (AdSV), multidrug usage, 51
Premarin InjectionÕ, 1630 1499 Naranjo causality algorithm, 54
protamine, 1632 Adult=acquired respiratory distress syndrome Naranjo’s classification, 47
sodium caprylate, 1630 (ARDS), 366, 1128 non-dose-related reactions, 47
SporanoxÕ, 1630 Advanced Care ViadentÕ, 898 nutritional factors, 50
tri-n-butyl phosphate, 1630 Advancement of Medical Instrumentation observational cohort, 53
use of, 1630 (AAMI), 3525 patient education, 54
zinc, 1632 AdvatabTM, ODT technologies in patient-related factors, 48
ADE. See Adverse drug events, 46. development, 1112 pharmacoepidemiology, 52
Adeno-associated viruses (AAVs), 3151 Adverse drug events (ADE), 46, 2243 types of systems, 52
Adenosine receptor agonist, 2809 medication errors, 46 pharmacokinetic dosing principles, 49
Adenosine triphosphate (ATP), 76, 1420 negative consequences of drug prevention of, 54, 1908
Adenovirus- and poxvirus-based vectors, misadventures, 46 reporting systems, 52
advantages of, 3910 Adverse drug event (ADE) risks, 1907
Adenoviruses, attenuated, 1612 assessment and identification, 1908 screening methods, 52
Adequate quality system, definition of, definition, 1907 change in medications, 52
3076 delirium, 1910–1911 computer report tracking systems, 52
Adhesion drug–food interactions, 1911 narrow therapeutic range drugs, 52
and cohesion, testing methods of, drug interactions, 1911–1917 tracer drugs, 52
2926 falls, 1909–1910 specific medical syndromes, 54
initial stage of, 2926 medication classes causing, 1908, 1912 type A reactions, 46
particle–wall interactions, 1538 symptoms associated with, 1908, 1910 type B reactions, 46
surface area, 1538 Adverse drug reactions (ADRs), 46 classification systems of, 46
Adhesional wetting, 3585 advantages of herbal therapies, 51 hypersensitivity reactions, 46
Adhesive age-related alterations, 48 immune-based reactions, 46
characteristics, 2667 compatibility and stability issues, 48 type I reactions, 46
hydrophilicity of, 2927 in children, 48 type II reactions, 46

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I4 Index

[Adverse drug reactions (ADRs)] [Aerosol] [Affinity chromatography]

type III reactions, 46 emulsification in, 3591 mobile phase in, 532
type IV reactions, 46 fine particle fraction (FPF) of, 2114 for separation
type C reactions, 46 fire extinguishers, 949 of antibodies, 532
type D reactions, 46 foam, 997 of enzymes, 532
types of, 1908 formation from ultrasonic nebulizers, of glycopeptides, 532
Adverse skin, classification, 1315 2105 of hormones, 532
Advertising and Promotional Labeling Staff generation, 2093, 3854 of metal binding amino acids, 532
(APLS), 63 based on the piezoelectric crystal, 3855 of peptides, 532
AdvilÕ, 3335 characteristics, 3858 of viruses, 532
Aero-FlowTM automated powder flowability control, 2086 stationary phase in, 532
analyzer, 3290–3291 mass median aerodynamic diameter AffmetrixÕ (Super-Array’s Oligo GEArray),
Aero-Flow rotating drum tester, flow (MMAD), 3856 2796
characteristics in, 3276 for inhalation, 2092, 3854 AftachÕ, 1254
Aerobic metabolism, 347 mechanism, 3854–3855 Agar-agar, advantages of, 2792
Aerobic organisms, growth of, 131 size, 3858 Agar plates, 2303
Aerobiologic contamination, 2307 uncontrolled vs. on-demand, 2086 Agarose hydrogels of, 2029
Aerobiologic samplers, comparison, 2310 output from nebulizers, 2098 Age-related macular degeneration (AMD),
Aerobiologic sampling, 2304 oral, 997 2441
methodologies particle size analysis Agglomerates
gravity settle plate, 2307 by inertial impaction, 2282–2283 disintegration of, 34
volumetric air sampler, 2307 by optical methods, 2282 drug-layered, 1777
procedures, 2307 particle size quantification, 2096 structure of, 1776
AeroChamberÕ, 997 performance testing, 2087 Agglomeration and coating
AerodoseÕ, 2705 pharmaceutical, 2092 materials for, 1775–1776
AeroDoseTM, 2112 properties, 2093 Agglomerative mechanisms
characteristics, 2112 salbutamol, 2100 coalescence, 2258
components, 2112 space, 997 layering, 2258
operation principle, 2112 surface, 997 nucleation, 2258
performance comparison, 2116 therapeutic inhalation, 3856 Aggrecan, 2436
Aerodynamic diameter, 2704 by ultrasonic nebulization, 3854 Aging
theoretical calculation of, 1799 Aerosol flow reactor dissolution characteristics on, 1861
Aerodynamic particle size assessment, 2087 biocompatible organic solvent for, 2387 physiologic changes with, 1905–1906
Aerodynamic pressure, 2095 hollow spherical morphology, 2387 Agitated batch driers, 3892
AeroEclipseTM key parameters, 2387 Agitator, for evaporation of viscous
breathing patterns, 2110 particle size distribution, 2387 materials, 3880
components, 2110 scanning electron microscopy (SEM) Agitator mixer, types of, 2976
performance comparison, 2116 micrographs, 2387 Agochloro fluorocarbons (CFCs), use of,
Aerosol Aerosol isolation suite, 2883 2980
aerodynamic diameter of, 2588 Aerosol particles, surface roughness of, Agriculture, excipients for, 1638
canister, 2277 1800 Ahrous carbamazepine, determination, 4109
characteristics, 2280 Aerosol photometer, 2174 Air attrition mills
strength, 2280 Aerosol products impinging jet mills, 2347
containers, 997 bioequivalence studies of, 3094 loop=spiral mills, 2347
definition, 949, 2092 spatial distribution of, 3094 particle size, 2347
delivery systems, 1283, 2092 Aerosol solvent extraction system, for types of, 2347
dosing reproducibility, 1283 microparticles preparation, 3575 Airbag gas generation technology, 1217
formulation stability, 1283 Aerosol sprays, 2282, 3971 Airborne contaminants, 2171
low drug mass per puff, 1283 Aerosol valve, operation, 2275 Air-classifying mill, 2347
low system efficiency, 1283 Aerosolization, 2080, 2093 Air-filled capsules, use of, 423
dosage forms, 997, 2109 Aerosolized solvent extraction system Air filters, 3888
deposition (ASES), 3570 asbestos in, 3888
factors, 2093–2094 AERxÕ, 2705 cellulose in, 3888
mechanisms, 2094 AERxTM, 2110 centrifugal operations, 3888
distribution by components, 2110 design, 3888
geometrical standard deviation (GSD), drug distribution in lungs, 2111 operation, 3888
2100 inhalation flow rates, 2111 testing of, 3888
mass median aerodynamic diameter metered dose inhalers (MDI) and, 2111 Air filtration theory particle-collection
(MMAD), 2100 performance, 2111, 2116 mechanisms, 2174
droplet size within, 3854, 3856 Affinity chromatography Airflow
drug classification conventional, 2175
delivery efficiency, 2081, 2098 bioadsorption, 532 laminar, 2172
particle size distribution, 2098 bioaffinity, 532 equipment
dry powder, 2085 immunoaffinity, 532 laminar, 2171

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I5

Air, humidity determination, 3884 [Alginate] Alpha-1 antitrypsin (A1AT)

Air ionization chamber, 3086 hydrogels, 2029 for hereditary deficiency, 2737
Air-jet nebulizers, 3854 in islet transplantation, 190 human plasma, 2737
Air–liquid interface, polysorbate at, 1429 molecules, 1186 recombinant human, 2737
Air–liquid interfacial degradation, 1428 nanoparticles, 1186 Alpha-chymotrypsin, precipitation of,
AiromirÕ, 997 wound contact layer, 1031 3578
Air-purifying respirator, 2884 Alginate-HEMA graft copolymer, 2336 Alpha cyclodextrins, 682, 687
Air samplers, volumetric, 2308 Alginate-polylysine encapsulated pancreatic Alpha decay process, 3084
Air–water vapor mixture, properties, islets, 2336 Alpha-folate receptor, 1328
1441, 3883 Alginic acid, 1883 Alpha-globulin protein, 3030
Airway diseases, treatment of, 2092 Algorithm of Fedorov, 2460 Alpha-1,4 glycosidic bonds, 682
Airway, human, 2093 Algorx, 1209, 1217 Alphahydroxy acids, 1316
Akaike’s Information Criterion (AIC) value, Alimentary canal, enzymatic activity in, Alpha-interferon, 1078
2766 1242 buccal delivery of, 1078
AL-6XNÕ (UNS N08367), 791 Aliphatic amines, titration, 3756 hepatitis treatment, 1078
Albumin, 1905 Aliphatic and aromatic monomers, surface- HIV treatment, 1078
drug delivery system, 1644 eroding polyanhydride polymers Alpha-keratin, solvation of, 13
human or bovine serum, 1613, 1616 preparation, 184 Alpha-lactose monohydrate, granulation of,
in hydrogel preparation, 190 Aliphatic polyanhydrides, preparation 746
in vivo glycosylation, 3037 of, 184 Alpha[-(2-piperidyl) beta-3,6-bis-
microspheres, 1138 Alkaline ester hydrolysis, 2042 (trifluoromethyl]-9-
delivery potential of, 1143 Alkaline hydrolysis process in bones, 406 Phenanthrenemethanol, 2221
5-fluorouracil, 1099 Alkaloids, 246–256, 2911 Alpha-tocopherol, 3347
protease-activated biodegradation, 1099 based on terpene skeleton, 254 metabolic pathway, 146
nanoparticles, 2333 biosynthesis of, 253 AltaceÕ, 1255
pasteurization of, 3999 derived from Alteplase protein, 266
synthesis of, 3029 nucleotide precursor, 255 Alternate chromatographic and
Albumin–acacia coacervates, 2333 other precursors, 256 immunological techniques, 3045
Albumin and globulins, separation of, 3028 phenylalanine and tyrosine, 248 Alternating current
Albumin bound drugs tryptophan, 251 polarography=voltammetry, 1497
protein binding of, 3036 ergot, 247 Alternative endotoxin test, 3058
Albuterol sulfate, 2112 oils extraction, 3902 Alternative medical practices, complementary
Alcohol, 998 steroid, 252 and, 67
and diazepam (Valium), 1043 Alkeran, 3359 Alternative medicine, 2902
as disinfectant, 1042 Alkoxyimino side chain, 2147 courses, 68
as recreational substance, 1042 Alkyl–oxygen fission (Al), 2040 definition of, 66
as solvent, 1042 nucleophilic substitution of carbon in, prevalence of, 2903
as vehicle in pharmaceutical formulations, 2040 products, 2903
1042 Alkyl chain anesthetics, 1059 regulation and oversight, 68–69
Alcohol and aldehyde dehydrogenases, in Alkyloxynol-741, 1354 limitations of, 69
drug biotransformation, 312 Allegra, 2422 resources, 80
Alcohol dehydrogenase (ADH), 1042 Allergenic extracts use of, 68
Alcoholic hepatitis, 75 aseptic filtration, 1267 Alternative reference methods, validation of,
Alcoholic solvents, inclusion of, 921 for diagnostic or therapeutic purpose, 2830
Alcoholism, 1042 1267 Aluminum acetylacetonate, 2928
Aldehyde dehydrogenase-2, deficiency of, sterile concentrates, 1267 Aluminum adjuvants, 3914
1019 Allergic contact dermatitis, 1315 in diptheria–tetanus–pertussis (DTP)
Alefacept, in psoriasis treatment, 275 Allergic reaction, 46 vaccines, 3915
Alendronate, absorption of, 1017 adverse skin, 1315 in human vaccines, 3915
Aleve, 2421 IgE-mediated reaction, 46 Aluminum chloride
Alexanderwerk Inc. roller compactor immunoglobulin E, 46, 49 as astringent, 805
with horizontal twin feed screw system, improvements in documentation of, 54 Aluminum containers
3173 Allergic-type reactions, 1616 corrosion of, 2274
sizing unit, 3174 Allium sativum (garlic), 72 manufacture
Alfacalcidol, 3348 Allogeneic red cell transfusions, 359 by deep-drawing process, 2274
Alginate Allopathic medicines, 2901 in metered dose inhalers (MDIs), 2274
applications, 2030 Allura red AC, 660 types
drug delivery system, 1644 Allylestrenol bulk drug substance and tablets, cut-edge can, 2274
in hydrogel preparation, 190 purity test for, 546 rolled-edge can, 2274
dressings Allylisopropylacetureide, 3435 Aluminum, corrosion resistance of, 793
calcium, 1031 Alpha-acid glycoprotein (AAG), 579, Aluminum ferrule, 2274
for soluble wound packing, 1032 1905–1906, 3028 Aluminum hydroxide gel, 1017, 1888
in wounds, 1031 Alpha-adrenoceptor antagonist, 2808 Aluminum magnesium silicate, 1889
wet integrity of, 1032 Alpha-amino acids, polymer of, 2717 Alveolar capillaries, endothelium of, 1280

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I6 Index

Alveolar cell monolayers AmiconÕ, 3028 Amorphous solids

type I, 1281, 2732 Amidated pectin beads, 1237 humdities for, 4050
type II, 1281, 2732 Amides, hydrolysis of, 2043, 3001 isotherm analysis
Alveolar cells, types of, 1281 Amikacin, plasma concentration profiles for, ambient temperatures, 4058
Alveolar epithelium 3954 as temperature function, 4060
alveolar cells, 1281 Amino-beta-lactams, 942 viscoelastic properties of, 4061
composition of, 1281 Amino acid-bonded transporter, 2721 water vapor absorption, 4052, 4058
definition of, 1281 Amino acid Guggenheim, Anderson, and deBoer
measurement, 1281 absorption of, 2721 (GAB) equation, 4052
Alza Corporation technology (MacrofluxTM), conjugation, 320 Amorphous state, drugs and excipients in,
1319 in domestic animal species, 3963 4108
AlzamerÕ, 186 reactions, 320 Amorphous sucrose, 2379, 4104, 4108
AlzetÕ osmotic pump detection of, 542 Amorphous system
applications of, 1093 in mammalian species, 320 freeze concentration in, 1809
cross-sectional view, 1095 methionine, 76 glass-transition temperature of, 1821
implantable or insertable CrDDS, 1093 and peptides, 1528 Amoxicillin, 2034
osmotic pressure-activated drug-delivery Aminobutylethylisoluminol (ABEI), 2057 as amoxicillin trihydrate, 3957
system, 1095 Aminoglycosides, 1229, 2634 penicillin allergy, 2255
semipermeable membrane, 1093 antibiotics, polar drugs, 3952 pharmacy software program, 2255
Alzheimer’s disease, 150 renal elimination of, 2636 weight on bioavailability of, 3957
treatment Aminomonosaccharide, 73 Amoxicillin trihydrate, 700, 1864
by acetyl-L-carnitine, 2440 Aminophylline tablets, 3964 intramuscular injection, 3956
by docosahexaenoic acid (DHA), Aminopyrine, 2034 and prednisolone, 3959
2440 Aminosalicylic acid salts, bioavailability of, Amperometric detector, 535, 1524
Alzheimer’s-type dementia, 73 3181 Amperometric principles, 1508
Amanita phalloides, 75, 76 Aminosyn II, 984 Amperometric titrations, 3765
Amaryllidaceae alkaloids, 248 Amiodarone hydrochloride, 3360 applications, 3766
biosynthesis of, 245 Amitriptyline, 2146 oxidation–reduction titrations, 3766
Amber container Amlodipine besylate, in tablets, 547 precipitation titrations, 3766
destabilizing effect of, 2862 Ammonia gas sensor, 1508 curves, 3765
disadvantage of, 2864 Ammonium oxalate, 3435 mirror image curve, 3765
Amber glass Amniotic fluid, 2631 polarographic methods, 3765
advantage of, 2509 Amorphous (glassy) materials procedures
stabilizing effect of, 2862 and crystalline materials, 2079 ion-combination reaction, 3764
Amberlite IRP-64Õ, 2126 formation, 85, 2078–2079, 2940 oxidation–reduction reaction, 3764
Amberlite IRP88Õ, 3561 glass transition temperature of, 2079 with two polarized microelectrodes, 3765
AmbisomeÕ, 1128, 3362 physical and chemical stability of, 2079 Amperometry, 1523
Amebocytes, osmotic lysis of, 3057 Amorphous and crystalline systems, Amphetamine
Ameliorate brain injury, 363 characterstics of, 86 abuse, 1044
American Association of Mechanical Amorphous compounds, 399 effects, 1045
Engineers (ASME), 2238 Amorphous dispersion, 83 enantiomers of, 2143
American Association of Pharmaceutical Amorphous drug delivery systems, 87 heterochiral, 2156
Scientists (AAPS), 1572 Amorphous drug powder, 86 prevention efforts and agents, 1045
American Diabetic Association (ADA), Amorphous excipients, 1648, 1822 racemic, 2156
2817 Amorphous hydrogels, in wound dressing, Amphipathic surface active molecules, 638
American Herbal Pharmacopeia, 2904 1030 Amphipathic surfactants, micellization of,
American Herbal Products Association Amorphous lactoses, 3230 638
(AHPA), 69 Amorphous pharmaceutical materials, Amphiphilic block copolymers, self-assembly
American Institute for Iron and Steel 399, 400 of, 1334
(AISI), 789 applications, 83–84 Amphiphilic polymers, 2222
American Inventors Protection Act, 2619 characteristics, 84–86 Amphiphilogels, 1062–1063
American Medical Association (AMA), definition, 86–87 Ampholytic surfactants, 3586
55 description, fundamental, 86–87 AmphotecÕ, drug products, 1643
American National Standards Institute, Inc., examples of, 85 Amphoteric compounds, solubility of, 926
(ANSI), 3525 molecular mobility, 88–89 Amphoteric drugs, 3957
American or western ginseng degree of, 86 fluoroquinolones, 3957
(Panax quinquefolius), 73 molecular motions in rifampin, 3957
American Pharmaceutical Association temperature dependence of, 400 tetracyclines, 957
(APhA), 2417 occurrence, 83–84 Amphotericin B, 2478, 3362
American Society for Quality, 3077 polyamorphism, 89–90 Amphotericine, antimycotic, 1128
American Society for Testing Materials properties of, 83, 86, 90 Ampicillin, 2938
(ASTM), 1108, 2238 solubility ratios of, 3314 antimicrobial efficacy of, 1190
American Society of Rheology, 3128 stability, 86–88 carbonyl resonances in, 2378
Ames assay, 1413, 2192 Amorphous polymers, 1743 elimination, 3965

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I7

[Ampicillin] [Analytical procedure] Anhydrous carbamazepine, 827

FTIR spectra of, 2378 linearity of, 95 polymorphs, nomenclature for, 847
powder XRD patterns of, 2378 for manufacturing and release of drugs, Anhydrous cholesterol, crystals of, 823
sodium and potassium salts of, 3181 96 Anhydrous metronidazole benzoate, 2940
therapeutic activity of, 1190 minimum range for, 95 Animal care programs, evaluation of,
trihydrate, 13C NMR spectra of, precision of, 96 125
2378–2379 quality control chart, 111 Animal-derived materials
Ampicillin-loaded nanospheres, 1190 quantitation limit (QL) of, 96 in drug products, 1641
Amplification, definition of, 84 range of, 95 egg phospholipid, 1641
Ampoule tightness tests, 3534 recovery deviations, 104 egg yolk lecithin, 1641
with poststerilization vacuum, 3539 relative standard deviation (RSD), 95 Animal drug use clarification act, 3992
Ampoules, 1976 reversed phase (RP), 98 Animal drugs, US regulation of, 3984
Amprenavir, 3336, 3349 robustness of, 96 food additives and drugs, distribution of,
dose of, 3349 sophisticated, 2830 3984
oral bioavailability of, 3349 system suitability test, 96 Animals
Ampuls thin layer, 98 in drug development, 114
constituents, 949 validation characteristics, 97 in drug safety testing, 117–124
definition, 949 applied, 98 acute toxicology, 118
Limousin’s rules, 950 by chromatographic separation, 98 antigenicity testing, 122
Amygdalin, 1508 peak purity investigations, 98 carcinogenicity, 121
Amyloid fibrils, 282 specificity, 97 mutagenicity, 120–121
Amyloidosis (AL), 282 specificity vs. selectivity, 97 primary irritation, 121
Amylopectin, 1885, 3476 types of, 94 principles, 117–118
branched, 3477 Analytical reference standards, 109 reproductive toxicology, 119–120
Amylose, 3476 Analytical solutions, stability of, 96 subchronic and chronic toxicology,
linear, 3477 Analytical ultracentrifugation 118–119
American Nurses Association (ANA), 55 particle density, 2397 use of, 1950
Analgesic drug (paracetamol), tablet particle size, 2397 Animal health management, effectiveness of,
formulation for, 1667 sedimentation rate, 2397 3982
Analgesics, 1077, 2435 Anaphylactic reaction, 47, 343 Animal models, 1661
buprenorphine, 1077 in paclitaxel (Taxol), 47 in drug discovery, 114–115
flurbiprofen, 1078 to surfactant Cremaophor EL, 47 for exciepient testing, 1661
ketobemidone, 1078 Anaprox, 2421 IPEC-America model, 1662
morphine, 1077–1078 ANDA. See Abbreviated new drug testing in humans, 1662
Analog-to-digital converter (ADC) board, application. route of exposure, 1661
3688 Andersen cascade impactor, 2283 Animal pharmacokinetics, 2494
resolution, 3688 Anderson–Darling statistic, 3507 Animal proteins, 267
sampling rate speed, 3688 AndrodermÕ system, 1084 Animal toxicity data extrapolation,
Analogue (N0915) Anesthetic agents 1424–1426
skin permeation enhancer, 1316 conception of, 2901 Animal toxicology
structure of, 1316 use of, 901 in cosmetic safety, 804
Analysis of variance (ANOVA), 3492 Anexothermic process, 1172 studies, 3065
dosage forms, 174 Angina Animal wax
null hypothesis, 3494 acute attacks, treatment of, 1076 lanolin, 4066
principle, 3495 pectoris, 1287 spermaceti, 4066
for testing equivalence of means, 3492 treatment of, GTN in, 1075 Anion-selective liquid-membrane electrodes
Analytical chemistry, 3016 Angiodema, cause of, 1616 preparation of, 1512
quality control, 2, 1371 Angiogenesis stimulating protein, 1328 Anionic dendrimers, 883
stability-indicating assays, 2, 1371 Angiography procedure, 344 Anionic emulsifiers, 3261
voluntary temporary interdepartmental Angioplasty procedure, percutaneous Anionic heparin, 600
transfers, 2, 1371 transluminal coronary, 346 Anionic perfluorinated surfactants, range of,
Analytical information systems, 734 Angiotensin converting enzyme (ACE) 1061
Analytical procedure inhibitors, 2722 Anionic polymerisation, of
accuracy of, 95 Angle-of-repose plate, of pharmaceutical alkylcyanoacrylate, 1184
analytical variability vs. specification powders, 3282 Anionic polymers, in sustained-release solid
limit, 97 Anhydride, hydrolytic reactivity of, 2043 dispersion preparation, 778
comparative test for, 2830 Anhydroglucose unit, 4059 Anionic surfactants, 14, 3586
detection limit (DL) of, 96 hydroxyl groups on, 4059 sodium cholate, 600
for drug development, 96 starch calcutation, 4059 Anise
gas, 98 Anhydrous a-lactose constituents, 1764
coefficient of correlation, 95 by a-lactose monohydrate dehydration, natural flavoring agents, 1764
homoscedasticity, 100 3682 waters, 994
investigation of, 105 Anhydrous calcium oxalate Anisotropic forces, effect of, 848
ion, 98 dehydration, 3732 ANN. See Artificial network.

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I8 Index

Annular shear cell, 3288–3289 Antibody-directed prodrug therapy (ADEPT) Antigenic peptides, 3910
for friction of pharmaceutical powders, drug method, 1146 Antigen positive cells, 1141
3289 Antibody drug conjugates, coupling methods Antiglaucomatous agents, 1195
Anodic oxidation, 1490 for, 1134 Antiglobulin, 277
Anodic stripping voltammetry (ASV), 1498 Anti-drug antibodies Anti-Helicobacter agents, 1253
Anorectal tissues, 1298 calibration curve, 1574 Antihemophilic products, 277
Anorexia nervosa, 2871 definition of, 1574 Anti-HER2 immunoliposomes, 1333
ANOVA quantities, calculation of, 3495 positive antibodies, 1574 Antihistamines OTC
ANOVA table validation, 1574 Allegra, 2422
calculation of, 3496 Anticancer agents, 117, 702, 1408, 1610 Claritin, 2422
construction of, 3495 delivery of, 1143, 1328, 1332 Zyrtec, 2422
Antacid tablets, formulation, 2406 Anticardiac myosin antibody, 1150 Antihyperlipidemic agents, 2922
Antacid testing procedures, 2419 Anticholesterol compound, experimental, Antihypertensive treatment, 364
Anterior superior iliac spine, 2646 2940 Anti-idiotype antibodies, monoclonal,
Anthelmintics, dosage of, 3960 Anticholinergic drugs, 1017 3912
Anthracene, irradiated crystals of, 3546 Anticholinergic effects use of, 1137
Anthracyclines, 1192 impairment in self-care capacity, 1395 Anti-idiotype vaccines, uses of, 3912
Anthraquinones, 2911 in drugs, 1393 Anti-immunoglobulin E (Anti-IgG)
plant-derived, 232 memory impairment, 1395 aerodynamic size distribution, 2738
Anthraquinones and xanthones, Anticoagulant drugs, 72, 237 aerosol delivery, 2738
biosynthesis of, 231 Anticoagulants, 2911 synchronization of optimum breathing,
Antiadherents, 1733–1734 absorption of, 1017 2738
Antialiasing filter, signal conditioning of Anti-ED-B fibronectin, 1333 Anti-inflammatory agents, 244
amplifier, 3687 Antiepileptic agents, clinical trials of, 2469 Anti-inflammatory arylacetic acids,
Antiallergic drug Antiepileptic drug development program, bioprecursors of, 3009
acidic 2469 Anti-inflammatory drugs, 544
ethanolamine salt of, 3179 Antifungal antibiotic griseofulvin, 1017 Anti-irritant properties of cosmetic
rank–order solubilities for, 3179 Antifungal drug (griseofulvin) formulations, 1317
Antiarrhythmic drugs, 1018 hard gelatin capsule formulation for, 1671 Antileukemia agents, 2469
Antiarrhythmic treatment, 588 Antifungal preparations, 2478 Antimalarial drug
Anti-asthmatic drugs, 1429, 1613 Antigen halofantrine, 1610
Antibacterial agent, selection of, 3972 altered processing of, 3914 lactate salt of, 3177
Anti-B-cell lymphoma, 1143 aluminum-based vaccines, 3915 mefloquine, 2861
Antibiotic agents cross-linked protein crystals, 3922 Antimalarial mefloquine, 3390
neomycin, 2479 delivery systems, 3919 Antimicrobial activity
in ototopical preparations, 2479 by microparticulate vaccine, 3920 enhancement of, 2992
polymyxin B, 2479 classification, 3919 spectrum of, 2985
Antibiotic formulations, gel-formers in, goal, 3919 Antimicrobial agent
161 live attenuated microorganisms systems, concentration effect of, 2987
Antibiotic powders, adsorption of, 35 3919 delivery of, 891
Antibiotics non-living microparticulate systems, half-life of, 3963
preparation of, 1646 3919 intramammary preparations, 3959
process of freeze-drying, 1646 determination of, 2048 local delivery of, 903–905
Antibodies competitive assays, 2048 parenteral preparations with, 3958–3959
in acute myoardial infarction, 1150 non-competitive assays, 2048 transfer of, 3959
anticardiac myosin, 1150 genetic stability of, 1144 various species of fish, 3963
carcinoembryonic, 1149 oral delivery of, 1195 Antimicrobial effectiveness test, 2785
charge-modified, 1153 overexpression of, 1327 principles of, 2786–2787
foreign antigens, 2048 production of, 2746 Antimicrobial mechanisms, 2987
glycoproteins, 1152 saponins, 3915 Antimicrobial preservative agents
in immunoassays, 2048 sustained release of, 3915 and adjunctive heat treatment, 1273
in immunotherapy, 1149 transcytosis mechanism of, 2728 constituents of, 1273
isoelectric point of, 1153 Antigen–antibody complex, 1567, 2049, inclusion of, 2991
monoclonal, 1149, 2048 3091 in ointment formulations, 3269
non-charge-modified, 1153 Antigen–antibody reaction, 1144 parenteral madministration, 1274
physiological conditions, 1152 Antigen-bearing cells, 1139 product serility, 1273
polyclonal, 1149 Antigen-binding ability, retention of, 1134, Antimicrobial resistance, 3981
preparation and manufacture of, 1133 1142 malaria and schistosomiasis, in humans
in radioimmunotherapy, 1149 Antigen binding capacity, 1132, 1134 pathogens
use of, 1132 Antigen binding regions, 1132 resistance of, 3981
Antibody-based drug delivery systems, Antigen density cells, 1140 Antimicrobial resistance monitoring system,
1134, 1144 Antigen density tumor cells, 1140 3983
Antibody-dependent cellular toxicity Antigen-encoding plasmid, delivery of, Antimicrobial Resistance Research
(ADCC), 1137 2746 Unit (ARRU), 3983

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I9

Antimicrobial risk management, resource for, Antipyrine Aqueous cleaning agents

3982 clearance interspecies scaling, 3965 commodity chemicals, 1584
Antimicrobial susceptibility, trends of, delivery of, 1299 formulated products, 1584
3982 Antireflux preparations, 1253 advantage, 1584
Antimicrobial therapy, 2635 Antisecretory drugs, nocturnal effects of, selection, 1584
Antimicrobials, use of, 3982 2872 types, 1583
Antimyosin Fab, 1151 Antisense oligonucleotides, 1184 water, 1583–1584
in acute myocardial infarction, 1151, Antisense probes, as therapeutics, 935 Aqueous coating solutions, 2330
1154–1155 Antisense therapy, advantages, 936 Aqueous fluid loads, biovalidation of, 332
in vivo administration, 1154 Antiseptic agents Aqueous metal ion solution, 1490
intravenous administration, 1154 for external ear canal disease, 2477 Aqueous nasal spray pumps, 1207
in myocardial necrosis, 1154 for otologic surgical prophylaxis, 2477 Aqueous pharmaceutical products, 329
negative-charge modified, 1153 Antiseptic chlorhexidine, 904 biovalidation of, 332
polymer modified, 1154 Antiseptic cottons, 950 Aqueous reaction mixtures, 3004
radioactivity of, 1153 Antiseptic gauzes, 950 Aqueous solubility, 84, 1245, 3002
Antimyosin immunoliposomes, 1159 Antiseptic pencils, 950 Aqueous versus non-aqueous cleaning agents,
cell morphology, 1164 Antisolvent addition rate profile, 865, 866, 1583
rhodamine labeled, 1162 868 Arabinofuranosylcytosine, 1141
silver grains, 1164 Antisolvent batch crystallization, Arachidonic acid metabolites, 3054
Antimyosin immunoscintigraphy, 1155 development of, 858 Arc path cutter, 2965
endomyocardial biopsy criteria, 1155 Antisolvent flow rate, calculation of, 867 Arch-breaking hopper design, of roller
Antineoplastic agent, 2036 Antisolvent precipitation compactor, 3168
clinical devolopment of, 2812 controlled precipitation, 2388 Area under plasma concentration time curve
water-insoluble, 3348 evaporative precipitation into aqueous (AUC), 1785
Anti-osteogenic sarcoma, adsorption of, solution (EPAS), 2389 drug absorption, 3710
1143 dissolution curves for, 2389 Arenediazonium salt, 1514
Anti-oxidants, effective, 356 drug-loaded organic solvent, 2389 Argentometric methods
Antioxidants Antisolvent supercritical fluid processes, determination
activity, quantification of, 150 3575 arsenates, 3761
addition of, 2927 preparation of polymeric drug halides, 3761
advantages of, 1274 microparticles, 3575 halogenoids, 3761
antimicrobial properties, 1625 insulin and catalase microparticles, mercaptans, 3761
butylated hydroxy anisole, 1625 3578 oxalates, 3761
propyl gallate, 1625 Antisolvent system, metastable zone for, phosphates, 3761
carotenoids, 143 864 sulfides, 3761
chelating agents, 1274, 2227 Antistatic spacers, 1543 Arginine salt, 3185
classification of, 142 Antitransferrin receptor, 1136 Argon–oxygen decarburization (AOD)
compatibility of drug with, 1625 Antitumor drugs, 1139, 2469 furnaces, 790
concentration range, 1626 release, 1777 Argon ionization detector, 198
constituents of, 1554 Antitumor effect, 1137 Argon plasma flame, 3373
cysteine as, 1625 evaluation of, 2808 Argonaut technologies, 3006
definition of, 139 Antitumor efficacy, 1329 Argonaute protein
dietary, 142 Antiviral agents, resistant to, 3981 Aromatic amines, 455, 1528
effect of, 2862 Antropyloroduodenal coordination, 2870 Aromatic hydrocarbons, biotransformation
exciepients, 1625 AnzemetÕ, 1256 of, 313
flavinoids, 143 AphA. See American Pharmaceutical Aromatic p-electron systems, 2161
free radical auto-oxidation reaction, 1274 Association. Aromatic water, 971, 994
frequency, 1626 Aphthous stomatitis, 1254 Arrhenius analysis, application of, 1690
hydrophilic, 142 API. See Active pharmaceutical ingredient. Arrhenius equation, 390, 400
lipid-soluble, 142 Apical luminal surface, 2724 Arrhenius law, 1689
maximum permissible amount, 1627 Apomorphine hydrochloride tablets, 922 Arrhenius temperature dependence, 86
natural, 142 Approximate dielectric requirement, Arrhythmia, 1014
oxidation–reduction potential, 1274 definition of, 808 Arsenic trioxide, advantage of, 2511
oxidation potential, 2227 Aptamers Arsphenamin
oxidative degradation, 2227 antibody–ligand interactions, 1575 molecular structure of, 1123
in pharmaceutical preparations, 1553 low-molecular-weight ligands, 1575 with thermotropic mesomorphism,
product stability, 2227 oligonucleotide sequences, 1575 1123
salts of sulfites, 2227 AquacoatÕ, 1737 Artemisia annua, 241
in small-volume parenterals, 1274 AquaFreshÕ, 902 Arthritis, anti-inflammatory therapy for,
synergists, types, 1625 AquaTar (Allergan Herbert), 1888 608
types, 1625 Aqueous=organic cosolvent systems, 682 Arthritis and Crohn’s disease, treatment of,
Antiozonants, 1467 Aqueous boundary (diffusion) layer, 1245, 2809
Antiperspirant, application of, 3821 2715 Articular cartilage replacement, 2029
Antiporters. See Exchangers. permeability coefficient of, 2716 Artificial cerebrospinal fluid, 364

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I10 Index

Artificial digestive systems (ADS), 2067 Asoxazepam, 1909 Atomic absorption spectrophotometry
class 1 drug, 2073 Asparagines, deamidation of, 1827 detection limits of, 3370–3371
efficacy of, 2074 Asparagus pea, 1180 ionization interference in, 3372
Artificial flavoring agents, 1765 Aspargine, deamidation, 283, 284 lamps in
Artificial lipid vesicles, 643 Aspartame, 1616 electrodeless discharge lamp (EDL),
Artificial neural networks (ANNs), 2399, Aspartame anhydrate (ASA), weight 3367
3435, 3650 fractions, 4112–4113 hollow cathode lamp (HCL), 3367
Artificial neurons, 2400 Aspartame hemihydrate (ASH), weight limitation of, 3367
Artificial radioactivity, 3082 fractions, 4112–4113 for metal analyses, 3367
Artificial salts, 950 Aspidosperma alkaloids, 253 in pharmaceuticals, 3367
Aryl ammonium salts, 1512 Aspirin operating principles of, 3367
AsacolÕ, 1254 active ingredients, 4106 sources of error in, 3372
Ascorbic acid, 145, 1825 chemical stability of, 42 viscosity effects in, 3372
in treatment of acetaminophen toxicity in degradation, 2382 Atomic absorption spectroscopy, 1490
cats, 3942 dissolution kinetics of, 389 Atomic displacements, in solids, 3544
Asepsis, definition of, 127 gastric irritation of, 944 Atomic force microscopy (AFM)
Aseptic coacervation processing, 612 hydrolysis, 2043 advantage of, 1794
Aseptic dispensing, 2136 instability of, 2043–2044 chemical nature of molecule, 876
Aseptic filling risks of, 2426 dendrimer molecules, 876
acceptance criteria for, 381 sorption isotherms for, 2373, 2375 dentritic structures, 875
batch manufacture, 381 teratogenic potential of, 2991 molecular structure, 876
broth fill contamination, 381 Aspirin-disintegrant mixtures, 2382 noncontact mode, 2395
incubation period, 381 Aspirin-Reye’s syndrome, 2420 pulse force mode, 876
microbiological growth media, 381 Assembled lipid bilayers, 1331 resolution of, 2395
processing capability, 381 Association colloids, 638 stiffness and adhesion properties, 876
sterility confidence level, 381 Association constant, inverse of, 3028 surface measurements, 2395
Aseptic manufacture, materials of, 328 Asthma, treatment by b-adrenergic agonists, Atomic number, 3082
Aseptic manufacturing activities, evolution 2090 Atomic structure, 3082
of, 130 Asthmatic attacks, incidence of, 1287 Atomization, 1799
Aseptic meningitis, 3059 chemical characteristics of materials, Atomizing air nozzle, 766
Aseptic pharmaceutical BFS technology, of 3632 Atopic dermatitis, treatment of, 2743
sterile liquid products, 383 physical characteristics of materials, Atrial natriuretic peptide, 364
Aseptic processing 3632 AtridoxÕ, 904, 3360
activity, 127 ASTM (American Society for Testing and AtrigelÕ, 3360
American Society of Health-System Materials), 3632 Atrisorb-FreeFlowTM, 3360
Pharmacists (ASHP), 46 Astringent pencils, 950 Atropa belladonna, 247
anaerobic medium, 135 Arterial blood flow, 2680 Atropisomerism, 2148
use of, 135 Atherosclerosis, 1158 AtroventÕ, 983
assurance for, 137 Atherosclerotic lesions, 1158 Attenuated total-reflectance (ATR)
caution, 128 carotid, 1160 spectroscopy, 3414
clean environment, 127 metabolic component of, 1158 for aqueous solutions analysis, 3414
environmental monitoring, 132, 133 muscle cells of, 1158 Attenuated total reflection–Fourier transform
gowning certification, 134 pathogenesis of, 1158 infrared (ATR–FTIR) spectrometer,
in health care industry, 127 in rabbit model, 1158 863
humidity in, 128 targeted, 1158 AUC (Area under concentration–time curve),
media fill program, 133 Athy–Heckel analysis 1892
microbial contamination, 128 on deformation mechanism, of drugs, Audit trials and electronic signatures
source of, 127 3643 comparison, 9
microbiological growth media in, 128 Atmospheric pressure chemical ionization in electronic batch record (EBR) system,
personnel monitoring, 134 (APCI), 1703 10
practice, survey on, 135 gas phase Auger-filling machine, capsules, 409
preparatory training, 134 ion chemistry, 1704 Auger cameras, 3087
principles of, 127–128, 132 process, 1704 in nuclear medicine, 3088
process simulation tests, in evaluation of, molecular ion fragmentation, 1705 Auger electron spectroscopy (AES), 2241
129 thermodynamically controlled ionization Auger electrons, 3085
manufacturing activities, 129 process, 1705 Augmented Acute Normovolemic
quantitative risk analysis, 128 Atmospheric pressure photoionization Hemodilution (A-ANHSM), 347,
in sterile material preparation, 127 (APPI), 3801, 3803 348
validation program, 127–138 Atmospherical corrosion, 793 Auranofin, polymorphs of, 2943
worst case situations, 128 Atomic absorption Auristilla
Asialoglycoprotein receptors, hepatic, 1330 components of, 3368 compound oil of Hyoscyamus, 950
Asian or Chinese ginseng (Panax ginseng), Atomic absorption burner medication of ear canal, 950
73 and inductively coupled plasma argon preparation, 950
AsmanexTM, 3252 plasma torch, 3368 Austenite–ferrite boundary, 790, 791

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I11

Austenitic stainless steels Avicel PH 200 Bacterial cell

application, 789 agglomerated irregular-shaped product, fixing and staining of, 2792
characteristics, 789 3230 membrane of, 3053
chemical composition of, 790 and cellactose, rheology of, 3286 pseudomonas, 1526
nickel in, 789–790 microcrystalline cellulose, 3230 surfaces, alteration of, 897
Autoclave chamber, 3535 Avidin bearing liposomes, 1145 Bacterial endotoxin, 3052
Autoclave treatment, 330, 1613 Avidin–biotin complex (ABC), 2053 clottable protein, 2293
AutohalerÕ, 997, 2109 Avidin–biotin reactions, 2053 control of, 2293
Autoimmune diseases, 71 Avidin–biotin systems, 2053 degree of specificity, 2293
Autoionization, of water, 385 dissociation constant, 2053 product-contact packaging components,
Autologous blood in immunochemistry, 2053 2293
donations, use of, 360 Avipox-rabies glycoprotein recombinant, production of gel, 2293
salvage system, 361 3910 pyrogen, 3053
volume, 361 Avitriptan sterile parenteral products, 2293
Autologous erythrocyte, 361 bioavailability, 2824 test, 3057, 3059
Automated dissolution systems, 740 drug development program, 2824 principles, 2789
Automated freeze-drying, 741 pharmokinetics of, 2824 threshold temperature, 2294
Automated metastable zone measurement, AvodartTM, 3348 validation and performance of, 2293
use of, 870 Azidothymidine (AZT) toxicity, 363 Bacterial enzymes
Automated multiple development instrument Azithromycin (Zithromax), 1399 hydrogel degradation by, 1237
(AMD 2), 541 Azo-coupling reactions, titration based on, matrix and hydrogels degradation by,
Automated potentiometric titrations, 1514 1237
equipment for, 1509 Azo-dextran gels, 2035 polymer degradation by, 1235–1237
Automated process development, type of, Azo-inulin gels, 2035 Bacterial glycosidases, 1255
3006 Azo-reductases, 1235 Bacterial infections, treatment of, 703
Automated sample handling and analysis, Azo dyes, 998 Bacterial KcsA channel, crystal structure of,
3006 Azone 4022
Automated tableting, 740 absorption enhancers, 32 Bacterial meningitis, treatment of, 2635
Automatic orbital welding effect of, 32, 1317 Bacterial metabolism, products of, 4043
components of, 2240 oral absorption enhancers, 32 Bacterial proteins, invasion of, 2723
square butt-welded tubing joints, 2240 structure of, 1316 Bacterial resistance, development of, 903
Automatic sampling, 2965 transdermal penetration, 32 Bacterial subcultures, 2303
methods AzoneÕ, 13, 1313 Bactericidal antibiotic concentrations, 903
discussion of, 2968 Bacteriostatic sodium chloride, 1006, 2637
Automatic transfer switches (ATSs), 1488 Bacteroides fragilis, 2479
Automatic welding machines, 4046 Babylonian clay tablets, 2901 Bag sampling, 2964
Automation software packages, 739 Bacillules Bagley plot, 1716
Automobile exhaust emissions, measure of, preparation, 950 Bailey’s model, 1539
2847 rod-shaped lozenges, 950 Baking, leaning process, 1586
Autoxidation Bacillus atrophaeus, 3526 Ball milling
autocatalytic reactions, 140 Bacillus calmette guerin, 2336, 3909 bead materials for, 2343
biological role of, 139 Bacillus coagulans, 327 efficiency of, 2343
of carotenoid, 145 Bacillus macerans amylase, 671 grinding process, 2344
definition of, 139 Bacillus stearothermophilus, 327, 2292 intrinsic parameters, 2344
free radical chain reactions, 140 Bacillus subtilis, 135, 379, 3519, 3888 agitator tip speed, 2344
inhibitors of, 140 Bacillus subtilis var. niger, 3512, 3519 bead loading, 2344
initiators of, 140 dry-heat sensitivity of, 3515 linear velocity, 2344
prevention of, 139 microbial death rate, 3526 mechanical force, 2343
rate of, 140 Bacillus xerothermodurans, 3516 nanoparticle size, 2343
reactions, 139 Bacitracin, 2700 residence time, 2343
phases of, 140 Back titration procedures for ethylenediamine scale-up performance, 2344
suppression of, 140 tetra-acetic acid (EDTA), 3760 spherical particles, 2343
of unsaturated lipids, 140 Back propagation algorithms, 2402 surface areas, 2343
Avalanching regime, 2354 Bacteria Balneum, 951
Average chart, 3500 colonic, 1615 Balsalazide, 1232
control chart limits for, 3501 gram-negative, 2791 Balsams
and subgroup size, 5301 live attenuated, 3919 definition of, 951
Avermectins types of, 4043 natural solutions of resin, 951
highly lipophilic substances, 3955 Bacterial hemolysin, 1139 Bancroft’s rule, 1554
veterinary preparations, 3971 Bacterial and parasitic infections treatment Bandages
Avian poxviruses, 3910 of, 3978 classification, 951
Avian species, 3943, 3953 Bacterial antigens, 3908 elastic, 951
in half-life of drugs, 3963 vaginal permeation of, 1343 inelastic, 951
Avicel PH 102Õ, 3566, 3675 Bacterial azoreductase enzymes, 942 ribbons of muslin gauze, 951

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I12 Index

[Bandages] BCS. See Biopharmaceutics classification Benzyl alcohol

semielastic, 951 system. advantage of, 1616
splint, 951 Bead inhaler technology, 1432 harmonized monograph, 1633
B- and D-tablet tooling Beagles, half-life of fentanyl in, 3973 preservatives of excipient, 1625
dimension, 3784, 3789 Bootstrap error-adjusted single-sample Bepex-Hosokowa Co. roller compactor,
heads of, 3786 technique (BEAST) 3174–3175
shapes, 3789 detection of NIRS, 3632 feed screw design, 3175
subcategories, 3784 non-parametric clustering algorithm, Bergey’s manual method, 932
Bar coding techniques, 555 3632 Bernoulli theorem
Barbiturate Beckmann’s freezing-point method, 3776 for fluid flow, 3862
alkyl, solubilization of, 3325 Beclomethasone dipropionate, 2107, 2980 pressure change determination, 3864
model T of, 1041 Becotide 100Õ, 2107 Bespak Piezo Electric ActuatorÕ, 3856
octanol–water partition coefficient of, Beer–Lambert law, 448, 3392, 3416 Bestatin, 2700
3325 for monochromatic radiation, 3416 Beta-adrenergic antagonists, 116
synthesis, 1041 Beer’s Law, 305, 864, 1971, 3367 Beta-adrenergic agonists, 1220, 2090
Bard Inspiron mini-neb, 2097 applicability of, 3461 as clenbuterol, 3942
Barium sulfate deviation from, 3461 Beta-blockers post-MI, 1922
characterization, 2379 Beeswax, 2330 Beta-carotene, 656
precipitation, 821 as coating material, 4072 Beta-carotene prooxidant activity,
Barwell ram extruder, 1726 insect wax, 4066 pathway of, 145
Basal gastric acidity, circadian rhythm of, as polishing agent in sugar coating, 4066 Beta-cyclodextrin, 459, 682, 689, 1258, 3328,
2872 as stabilizer of w=o-emulsions, 4066 3329
Basal secretion level, in human, 2749 as stiffening agent, 4066 chemical structre of, 674
Basic local alignment search tool (BLAST), white and yellow, 4066 17b-estradiol, 148, 2442
3151 Bench-scale device, 2352 biotransformation of, 778
Basolateral cell membrane, 1244, 2715, Bench-scale filling machines capsules, 410 Beta-Galactosidase expression, in tissues of
2722 Bending vibrations, 3406 mice, 3913
surfaces, 2724 Benign prostatic hyperplasia (BPH) treatment Beta-glucosidase, 1508
transporters of, 2721 of, 2807 Beta-Glucuronidase, 1341
Basolateral plasma membrane proteins, Benoxaprofen, polymorphic forms of, 17b-hydroxysteroid dehydrogenase, 2443
2720 3300 Beta-interferon preparation, antiviral activity
Bass larvae feeding of, 2330 Bentonite, 994–995, 1721, 1888 of, 2674
Batanopride hydrochloride BentylÕ, 2253 Beta-lactam antibiotics, 1256, 1244, 2722
aqueous solution of, 390 Benzaldehyde, 1633 b-lactamase, production of, 4100
decomposition pathway for, 390 Benzalkonium chloride, 292, 901, 2992 b-receptor blocking agents, 1287
Batch-forming process Benzamide crystal, intermolecular b-sheet conformation, 1311
process method of, 2233 interactions in, 617 Betamethasone valerate and miconazole
sizing rollers, 2233 Benzene nitrate, in cream preparations,
Batch crystallization as coating solvent, 1289 547
control of, 858 and dichloromethane, use of, 2996 Bevantolol, absorption of, 1017
operation of, 858 Benzimidazole anthelmintics, veterinary Bexarotene, 3336
optimization studies, 865 drugs, 3941 Biaxial shear testers
process control, 865 Benzisoquinoline hydrochloride, 3735 pharmaceutical powders characterization,
development of, 865 Benzo(a)pyrene and perylene, in 3289
Batch ovens, 3515 ethanol=water systems types
Batch production record, specification of, solubilization of, 3319 flexible boundary tester, 3289
1945 melting point of, 3319 mixed boundary testers, 3289
Batch validation, 554,556 Benzocaine, solubilization of, 3320–3321 rigid boundary tester, 3289
Baths Benzodent, 901 Bicarbonate buffer, 356
classification, 951 Benzodiazepine, 1909 Bifomyk gel, 1126
cold, 951 advantage of, 3359 Bifunctional reagents, 1135
external application of water, 951 antagonists, 1043 advantage of, 1136
hot=vapor, 951 binding, 3030 Bifurcation, femoral, 1159
tepid, 951 metabolites of, 311 Big BlueTM mouse models, 1414
therapeutic techniques, 951 Benzoic acid Bilayered skin equivalent, 1035
warm, 951 derivatives, micellar partition coefficients Bile
Bayesian adaptive control approach, 2812 of, 3325 constituents of, 24
Bayesian analysis, 2756, 2758 glutamic acid derivative of, 1146 hepatic, 24
Bayesian feedback approach, application of, preservatives, 1614 pH of, 24
2812 Benzophenathradine, 897 salts, 24
Bayesian inference criterion, 2405 Benzoyl benzoate, 779 absorption enhancers, 31
B-cell lymphocytes, 1137 Benzoyl peroxide, 3970 constituents of, 31
B-cell lymphomas, 1137 Benzphetamine, dissolution of, 3181 effect of, 15
B-cell lysis, 1137 Benzydamine, 1356 in stomach, 3593

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I13

Biliary excretion, 114 Bioanalytical assay, sensitivity of, 2922 [Biodegradable polymer(s)]
Bilirubin Bioartificial organs, design of, 612 erosion versus chemical changes, 177
neurotoxicity, 370 Bioassay, concept of, 438 hydrolysis of polymer chains, 176
serum concentration of, 3036 Bioavailability degradation rate of, 179
Bin blender. See Tote blender. absorption, rate of, 1892 as drug carriers, 176
Binary blend systems, 1430 definition, 1892 drug delivery systems, 177
Binary drug–carrier systems, 762 degrees of, 2942 examples of, 179, 180
Binary-phase diagram, 775 dosage forms, 1892 and glass transition temperature (Tg),
with eutectic formation, 775 generic drug product, 1892 179
Binder dispersion, degree of, 36 human test of, 2850 limitation of polyorthoesters as, 187
Binder solution, adsorption of, 36 measurement, 1893 mass transport rates of, 179
Binder surface tension, 36 pharmaceutical excipients, 1892 in matrix (monolithic) system preparation,
Binders, wet granulation process, 3658 physiological factors associated with, 940 181
Bingham bodies, 3132 testing, 1893 and melting temperature (Tm), 179
model, 3132 Bioavailability and bioequivalence studies, phosphazenes as, 187
rheogram of, 3130 1785 phosphoesters as, 187
shear strain rate for, 3130 during IND period, 1784 properties of, physicochemical, 179
Bingham-type plastic flow, 3604 new drug applications (NDA), 1784 polymer composition vs. physicochemical
pharmaceutical gums and hydrophilic orally administered drug products, 1785 properties, 179
colloids, 3604 systemic exposure measures, 1785 surface-erosion of, 181
property of, 3604 Bioburden testing heterogenous hydrolysis, 181
yield stress, 3604 analysis of bulk solutions, 380 in sustained release drug delivery
Binomial and normal distribution theory, and manufacturing process, 380 poly(D,L-lactide), 177
3491 non-sterile environment, 380 poly(D,L-lactide-co-glycolide), 177
Binomial distribution, 3490 polymer granules, 380 tailor-made dosage forms and delivery,
binary outcomes, behavior of, 3490 reduction=inactivation, 383 177
Bio-Cor, 1222 stages of liquid processing, 380 thermal properties of, 179
Bio-Dis apparatusÕ, 912 Biocatalyst, insoluble, 1508 Biodegradable polymer selection
Biovalidation cycles, 333 Biocatalytic membranes, 1508 factors affecting, 179–180
Biovalidation steam sterilization, principles, Biocatalytic urea sensor, 1508 homopolymers vs. copolymers, 179
325 Biochemical balances, maintainance of, 3041 and regulatory approval, 179
Bioabsorbable hemostatic agent, 155 Biocide, fraction of, 2988 Biodegradable sutures, moisture and, 177
Bioabsorbable polymers Biocompatible emulsifier, selection of, 342 Biodegradation definition of, 177
application of, 155 Biocompatible polymer, 1329 Biodegradation vs. bioerosion, 177
types of, 155 Biodegradable powder inhalation products, Biodressings
Bioabsorption, definition of, 155 415 types
Bioactivation mechanism, 1255 Biodegradable crosslinked polymer networks, biological dressings, 1034
Bioactive agents, chemical stability of, in drug delivery, 190 biosynthetic dressings, 1034
2380–2381 Biodegradable delivery systems in wounds, 1034
Bioactive dressings, in wounds, 1036 development, for proteins, 178 Bioequivalence study, 3989
Bioactive peptides and biodegradable delivery drug stability, factors affecting, 178 Bioequivalent drug products, 222
systems, 178 for synthetic analogs of luteinizing Bioequivalency
Bioactive wound management products, hormone-releasing hormone absorption rate, 1893
1036 (LHRH), 178 advantage of, 1893
Bioadhesion Biodegradable drug delivery blood concentration curve, 1893
biological synthetic and natural polymers in, 179 characterization, 1893
macromolecules, 1169 new approaches for, 190 disadvantage of, 1893, 1894
tissues, 1169 Biodegradable enzymes, 258 measurement, 1893
definition of, 1169 Biodegradable gel, 1031 model independent method, 1892
measurement of, 2668 Biodegradable hydrolyzed gelatin matrix, multidose, 1893
phenomenon of, 2667 904 single-dose, 1893
theory of, 2668 Biodegradable nanoparticles, 2384 statistical-moment theory, 1892
Bioadhesive–biological interface, 1172 Biodegradable polyesters, 1010, 1334 steady-state theory, 1893
Bioadhesive delivery systems, 1257 Biodegradable polymer(s), 1143, 1183 systemic circulation of dosage form, 1893
principles of, 1169 advantages of, 176 testing of, 1892
Bioadhesive dosage, use of, 1064 bulk degradation vs. molecular weight, therapeutic equivalence, 1892
Bioadhesive force, 1172 183 Bioerodible polymers, 177
Bioadhesive gels, 1064 classes of, 176, 180 Bioerosion, definition of, 177
Bioadhesive hydrogel formulation, 1064 hydrolytically labile polyesters, 176 Bioerosion-regulated drug delivery systems,
Bioadhesive polymer, 1253 classification of, 179 1099
Bioadhesive properties, thermal gelling common classes of, 182 drug-dispersed bioerodible matrix
polymers with, 1299 degradation mechanisms, 176 fabrication, 1099
Bioadhesives, residence time of, 2668 chemical changes to the polymer, 176 feedback-regulated system for, 1099
Bioaerosol samplers, 2310 erosion, 176 Bioerosion vs. biodegradation, 177

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I14 Index

Biofilm Biologics licensing applications (BLAs), 1782, [Biosensor potentiometric]

bacterial growth, 4043 1788 hydrogels in, 2030
definition of, 4043 Bioluminescence, 3401 hydrogen peroxide, 2030
Biogenesis of aconitine, 251 See also Chemiluminescence, 204 poly(vinylimidazole), 2030
Biogenesis of nocardicin A, 256 Biomarkers, 267–268 Biosynthesis of
Biogenetic theory, 228 screening use of, 3048 amaryllidaceae alkaloids, 245
Bioinequivalence, definition of, 2850 Biomedical drug analysis, 194 camptothecin, 250
Bioinformatic techniques, applications, 2492 developments in, 195 citrinin, 230
BioJect, 1217 gas-pressure liquid chromatography, 195 colchicines, 244
Biological dressings high-pressure liquid chromatography, desacetylcephalosporin C, 255
cultured, 1035 195 drugs, 228
natural, 1035 ultraviolet (UV), 195 emetine, 245
Biological fluids visible light absorption or emission, 195 ergot alkaloids, 247
analysis Biomedical trace analysis fatty acids, 229
fluorescence densitometry, 545 absorption photometric detectors, 199 flavonoids, 235
visual zone comparison, 545 for high-performance liquid furanocoumarins, 235
drugs in, 516 chromatography, 199 griseofulvin, 231
Biological Ghost Delivery Systems, 1335 detectability, 199 hydrastine and sanguinarine, 243
Biological indicator evaluator resistometer electrochemical detector, 199 isoflavonoids, 236
(BIER), 3525 fluorescence detector, 199 lanosterol and cycloartenol, 238
Biological indicators sterilization of, 3525 types of detectors for, 199 (S)-norcoclaurine and (S)-reticuline, 242
Biological license applications (BLAs) and Biomembrane characteristics, 2667 penicillin G, 255
stability information systems, 733 Biometric identifier, sort of, 2556 prostaglandins, 230
Biological matrices, drugs in, 1497 Biomolecular pharamacologist, 3047 protoberberine alkaloids, 243
Biological medicines, types of, 2830 Bionite lenses, 1221 pyrrolizidine alkaloids, 241
Biological membranes, lipid peroxidation of, Biopharmaceutic classification, basis for, quinine and quinidine, 251
150 1246 strictosidine, 247
Biological processes, metals in role of, 3367 Biopharmaceutic classifications system, 419, tropane alkaloids, 240
Biological product 3714 vinblastine, 249
growth factors (GFs), 270–271 category, 2817 Biosynthetic dressings, in wounds, 1035
hormones, 269 classes of drug substances, 167 Biotechnical and genetic engineering, 570
interferons, 269 implementation of, 926 Biotechnology
stability=degradation of, 262 in vivo methods, 167 drugs nature of, 2746
types of, 269 mechanism, 167 future of, 278
in United States, 1955 parameters, 926 history of, 259
Biological product categories, 268–269 Biopharmaceutic experimentation, 2821 industry maturation of, 2741
Biological safety cabinet (BSC), 2173 Biopharmaceutical therapeutic agents, products, 94
airflow patterns of, 2178–2181 1612 delivery of, 2741
applications Biopharmaceutics, 3648 research geography, 259
in industry, 2181 bioavailability, 222 technology of, 259–260
in pharmaceutical manufacture, 2181 bioequivalence, 222 Biotechnology-derived products, 1957
in pharmacies, 2181 definition of, 208 based on
in sterile testing, 2181 lackluster potency, 2, 1371 animal-source insulin, 1957
cabinet ventilation, 2179–2180 oral absorption, 2, 1371 insulin, 1957
classification, 2178 and pharmacology, 2, 1371 prospective harmonization, 1959
class I, 2178–2179 Biophase distribution model, 2804 safety evaluation, 1421–1422
class II, 2178–2179 first-order distribution rate constant, Biotherapeutic products, 3060
class III, 2178–2180 2804 Biotinylated antibodies, mixture of, 1145
containment and barrier techniques to, hypothetical effect–compartment, 2804 Biotinylated liposomes, binding of, 1145
2178 peripheral pharmacokinetic compartment, Biotinylation, 2053
in drug manufacture, 2178 2804 Biotransformation
laminar flow, 2178 Bioreactors types, 3905–3906 of aromatic hydrocarbons, 313
location and maintenance of, 2180 Bioresearch monitoring inspections, 3072 chemical exposure, 322
operating procedure, 2180 Bioresponsive drug delivery systems, 1100 conjugation reactions, 317
performance, 2182 cross-sectional view, 1101 drugs
pharmacy operations, 2178–2179 feedback-regulated system, 1100 arylacetic acid (indomethacin), 311
protection design features of, 2180, 2182 glucose-triggered insulin delivery system, arylpropionic acid (ibuprofen), 311
suitability, 2179 1100 definition, 310
Biological substances, monographs on, mechanism of, 1101 detection of metabolites, 311
2830 Biosensor potentiometric, 1525 electrophilic substances, 311
Biological systems acetylcholine, 2030 by enzymic process, 310
organic compounds in, 3090 cell-based, 1525 functional groups, 311
radioresistant, 3550 for detection of alcohols, 2030 by non-enzymic process, 310
Biological tissues, cavitation in, 3837 for enzyme immobilization, 2030 nucleophilic substances, 311

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I15

[Biotransformation] [Biotransformation pathways] Blood–brain barrier, 577, 2635, 2681

polar hydrophilic moiety, 311 of aromatic hydrocarbons, 313 Blood–milk barrier, 3959
reactive intermediates, 311 of epoxides, 313 Blood–vaginal epithelium barrier, 1355
in epidermis, 3968 of primary alcohols, 312 Blood capillaries, 2665
epoxides, isomeric phenols, 313 of secondary alcohols, 312 Blood cell growth factor, 270
by gut flora, 311 of unsaturated hydrocarbons, 313 Blood cell production, process of, 363
hydroxylations on aliphatic carbon, 312 Blood coagulation, 704
catechols, 314 oxidative enzymes in, 312 in acute myocardial infarction, 273
NIH shift, 314 phase I, 311 deficiency of, 272
induction phase II, 311 factors, 271–272
of Barbiturates, 322 reduction, 314 stages in, 3058
consequences of, 322 in sulfur, 315 Blood components, electrostatic interaction
of Dieldrin, 322 reversible, 311 of, 1332
inhibition BioValve, 1217 Blood drug concentration–time curve
of Cimetidine, 322 Biowaivers, 226 division, 169
consequences of, 322 BiphetamineÕ, 1771 dosage forms, 168
N-oxidation Bipolar neurons, 2681 Blood ethanol concentrations, 2639
in hydroxamic acids, 314 Birds Blow–fill–seal (BFS)
in hydroxylamines, 314 anseriformes, 3963 advantage of, 379, 747
nitroglycerin, 322 galliformes, 3963 air-borne contamination, 379
of nitrogen micronebulizer, 2097 aseptic processing technique, 378
azoreductase activity, 315 Birefringence effect, 446 automation of, 381, 383
in azo dye Prontosil, 315 Bismuth subnitrate suspension flocculation, broth fill contamination, 381
nitroreductase activity, 315 4125 broth filled units, 379, 383
oxidation Bisphenol A (BPA) preparation of coolant systems, 380
of divalent sulfur atoms, 314 polyphosphoesters from, 189 container contamination, 379
and lipophilicity, 312 1,4-Bis(hydroxyethyl)-terephthalate (BHET), for cosmetics, 378
of nitrogen, 314 190 dry heat sterilization process, 383
at saturated (sp3) carbon, 311 Bisphosphonate drug clodronate environmental contamination, 379
of sulfones, 314 bioavilability of, 2825 extrusion process, 380
of thioethers, 314 Bitter orange filling environment, 379
at unsaturated (sp2) carbon, 311 constituent of, 1765 filling mandrels, 378
pathways of, 311 essential oil, 1765 for foods, 378
properties of, 310 Black belts, definition of, 3079 high levels of sterility, 379
reduction Black body radiation integrity testing, 383
of halogenated hydrocarbons, 315 emissive power, 3874 leaking units in, 383
of sulfur, 315 heat transfer, 3874 localized air shower, 379
of reduction Stefan–Boltzmann law, 3874 for liquid product categories, 378
in carbon–carbon double bonds, 315 Black cohosh (Cimicifuga racemosa), machines, 380, 748
in nitrogen, 315 69–71 microbial contaminants, 379
stereoselective, 320 Black tea coolant systems, 380
stereospecific, 320 causes, 2438 raw materials, 380
in asymmetric centers, 320 consumption, 2438 microbiological monitoring, 378
hydroxylation of secondary carbon flavonoids for non-sterile medical devices, 378
reduction of ketones, 320 theaflavins, 2438 pharmaceutical, 378
in stereochemical inversion, 321 thearubigin, 2438 plastic containers, 378
of sulfoxidation, in thioamides, 314 Bladder tumor(s) polymer granules, 378, 380
of sulfoxide formation, 314 formation, 2776 process, 747, 748
sulfation, 318 preclinical findings of, 2779 routine microbiological environmental
of aromatic hydrocarbons catechols Blaze wavelength, 3397 monitoring, 379
terminal metabolites, 314 Bleeding, postoperative, 73 semiquantitative air monitoring, 379
Biotransformation pathways Blenders specification class for, 378
by enzymes in liver, 311 drum, 2355 spore contamination, 383
factors affecting, 311 tumbling, 2352 sterile pharmaceutical products, 380
hydroxylation, aromatic hydrocarbons, Blending regimens, 2352 sterilizing grade filters, 378
313 Blend specimens analyses of, 3492 sulfur hexafluoride (SF6) tracer gas, 379
metabolites of Blend uniformity surface monitoring, 379
amines, 312 history of, 2967 validation of
thioethers, 312 replacement of, 2968 clean in place (CIP), 380, 381
oxidation, aromatic ring systems, 313 Bleomycin, 1352 steam in place (SIP), 380, 381
oxidation and deamination of primary Blister packages, 1976, 2085 Blood flow variation, regional effect of,
amines, 313 Blister plasters, 951, 965 3822
cleavage of esters, 313 Blood-borne pathogens, 353 Blood glucose level, 116, 1428
of arene oxide intermediates, 313 Blood boundary layer permeability of, 2716 change in, 2751

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I16 Index

Blood level studies, dosage forms, 168 Bovine mammary gland milk production in, [Broth fills]
Blood, oxygen-carrying properties of, 353 3957 blow-fill-seal units
Blood partition coefficient, 2759 Bovine red cells, 355 acceptance criteria, 382
Blood replacement, principle of, 354 Bovine serum albumin (BSA), 2160, 2318, internal surfaces of, 382
Blood salicylate levels, increase of, 2991 2391 microbial contamination, 382
Blood substitutes, 335 release of, 1237 operator activity, 382
fluorocarbon approach, 335 Bovine spongiform encephalopathy (BSE), contamination, 381
Blood, transfusion, allogeneic, 335 1642, 3206 environmental contamination, 381
Blood versus plasma clearance, 3033 Bovine type 1 collagen, 1035 frequency and size of, 382
Blood vessel, component of, 1326 Bovine viral diarrhea virus (BVDV), 3999 incubation period, 382
Bluish–gray stains, 902 inactivation, 4000 machine recommissioning, 382
BMS-182994205, process development of, reduction, 4000 operational qualification of Blow-Fill-Seal,
2996 Bowman–Birk inhibitor, 2726 381
Body composition, differences in, 2634 Box–Wilson design, 2458, 2460 statistical probability of, 382
Bohdan automation, 3006 Brønsted–Lowry model, 385 sterilization, 382
Bohle concept, 745 Bradyarrhythmias, 76 Brown–Richards equation, for static flow,
Bohr’s planetary model, 3082 Bragg’s Law, 2940 3281
Boiler and pressure vessel code (BPVC), Brain-computed tomography scan, 364 Brownian motion, 1796, 3889
2239 Brain cells morphological change, 1803 advantages of, 2386
Boiling liquids Brain injury clinical management of, 366 of colloidal particles, 2386
heat transfer, 3872 Brain metastases, 1328 definition of, 640
in vertical tubes, 3873 Brain stem ischemia, 347 in dispersing medium, 2386
Boiling point elevation Brain tumor, treatment, 1328, 2322 pharmaceutical particles, 4119
Clausius–Clapeyron equation, 3771 Brass alloys, dezincification of, 784 stability of, 2386
colligative property, 3771 Bravais-Friedel Donnay-Harker (BFDH) Stokes–Einstein theory, 2386
definition, 3771 model, 847, 848 Brownian particles, 1797
ebullioscopic constant, 3771 Breast cancer Browning reaction, 2232
molal elevation constant, 3771 cells, 150, 1328 Brunauer–Emmett–Teller (BET) surface area,
non-volatile solute, 3771 primary, 1137 1793
osmotic pressure measurement, 3776 models, 1139 Brunauer, Emmett, and Teller equation, 4051
tonicity, 3776 patients application, 2372, 4051
Boltzmann constant, 816 genetic abnormality of, 264 non-polar gas adsorption, 4051
Bone survey of, 68 linear form, 4051
components, 2441 Breast carcinoma xenograft, 1139 modification of, 2372
infection, antibiotic formulations for, Breath-activated device, 1923 in surface area determination, 2372
162 Breath operated inhaler (BOI), 2277 for vapor adsorption, 4051
marrow BreathancerÕ, 997 Brush border
cytogenetic analysis, 120 Brevundimonas diminuta, 383, 1748 definition of, 2715
progenitor cells, 266 BrexinÕ tablets, 1258 peptidases, 2718, 2721
osteoporosis, 2441 BricanylÕ Turbohaler, 1540 Brushing, mechanical methods of, 900
Bone health, 2441 Brigham tea, 74 Bubble-point performance, 3902
nutraceuticals for, 2441–2442 Brilliant blue FCF, 660 Bubble drying process, 1429
g-linolenic acid, 2442 BrioÕ, 557, 558 Bubble point test, 1755
n-3-PUFAs, 2442 British Drug Regulatory Agency, 568 integrity test, 1755
soy isoflavones, 2441–2442 British Herbal Compendium, 2904, 2906 measurement of, 1755
Bone mineral density (BMD), 2442 British Herbal Pharmacopeia (BHP), 2904, microporous membranes, 1755
Bone morphogenic growth proteins (BMP), 2906 wetting fluid, 1755
275 British patent system, basis for, 2605 Buccal absorption, 21
Borosilicate glasses, 2514 British Pharmacopoeia (BP), 1753, 2212, 2857 for drug adminstration, 1071
chemical durability of, 2519 Brodie’s principle, 194 model, of drug, 1074
Borosilicate microfibers, 2173 Bromobutyl rubbers, 1475 Buccal adhesive
Borosilicate pharmaceutical glasses 4-bromo-1-naphthalenediazonium chloride, patches, 2670
compositions and thermal 1514 tablets, 2670
expansions of, 2515 Bromocriptine, 1356 Buccal administration, presystemic
Boston bioprobes, 935 Bronchial asthma, 1287 metabolism prevention, 1075
Botanicals, analysis of, 2851 Bronchial carcinoma patients, clinical trials Buccal bilayer devices
Boundary diffusion layer, 1495 with, 1140 components of, 1175
Bovine adenovirus type 3 (BAd3), 3912 Bronchial epithelial cells, human, 1141 drug-containing mucoadhesive layer, 1175
Bovine and porcine gelatins, alternative for, Bronchodilator, 2100 drug-free backing layer, 1175
420 terbutaline sulfate, 2980 Buccal bioadhesive captopril tablet, 1077
Bovine corneal opacity (BCOP) test, 1420 Bronchopulmonary tract, 2980 Buccal delivery devices
Bovine growth hormone (bGH), 2736 Broth fills bioadhesive polymers in, 2669
Bovine insulin, 460 aseptic processing capability effect, 381, 383 development of, 2674
Bovine leukemia cells, 1141 batch documentation, 382 geometric designs of, 2669

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I17

Buccal dosage forms, 2669 Bundesverband de Pharmazeutischen Calcitonin gene-related peptide (CGRP),
formulation development of, 2669 Industrie (BPI), 1982 delivery of, 2749
Buccal drug retention, duration of, 2667 Bungyo, 1980 Calcitrol, 3347, 3349
Buccal epithelium thickness of, 2665 Bupivacaine-free base, dispersions of, 3305 dose of, 3349
Buccal glands, 1072 Bupivacaine–HCl, dispersions of, 3305 Calcium alginate gels, 1883, 2317
Buccal homogenate studies, 2674 Bupivicaine, therapeutic concentrations of, formation of, 1878
Buccal insulin absorption, 2672 3038 Calcium alginate microcapsules, 2331
Buccal morphine, 1078 Buprenorphine, 1077 Calcium carbonate
Buccal mucosa absorption of, 1299 formation, 3732
carrier properties of, 2666 bioavailability of, 1077 tablets, 922
D-glucose and L-arabinose to extraction ratio of, 1077 Calcium-channel-blocking, 2150
absorption of, 1073 Burkitt’s lymphoma cell lines, 1143 verapamil in, 1192
isosorbide dinitrate in Burn wound healing, gel-formers in, 162 Calcium-chelating agents, 2720
absorption of, 1073 Burns Calcium gluconate injection, 1630
permeability of, 2665 management of, 1029 Calcium glycerophosphate (CGP), 700
presystemic metabolism in, 1076 treatment of, 1028 Calcium hydroxyapatite, 704
proteolytic activity of, 2674 Buserelin, 1078 Calcium pectinate, insoluble polymer, 1291
Buccal tablet absorption of, 17 Calcium phosphate, 2454
prototype for, 1289 bioavailability of, 1078 transformation of, 899
BuccastemÕ, 1254 in endometriosis treament, 1078 Calcium sulfate, stages of hydration, 2371
Buccoadhesive morphine sulphate tablets, treatment of hormone dependent tumors, Caliberation of NIR
1078 1078 by differential scanning calorimetry (DSC),
Buchner funnel, 1269 Business rules, implementation of, 2555 3437
Budesonide, 2107 BusulfanÕ, drug product, 1643 thermal gravimetric analysis (TGA),
anti-asthmatic agents, 1613 Busulfex, dose of, 3360 3437
nebulizing suspension, 2112 Butadiene rubber, 2275 Calorimeter, 393
Budesonide–PLA particles, 3575 Butane compression, 403
Buffer(s) advantages, 1212 isoperibol, 402
acetate, 1627 combustion engine, 1212 Calorimetric glass transition temperature,
adjusting agents, 1628 in needle-free injection, 1212 400
benzoate, 1627 Butorphanol tartrate, 946 Calorimetric methods, for drugs quality
citrate, 1627 Butt adhesion test, 1739 control, 403
components, crystallization of, 840 Butyl rubber Calorimetric techniques, thermal events,
definition of, 387 advantages of, 1276 394
ionic equilibria of, 387 water vapor permeation properties, 1276 Calorimetry
in pharmaceutical systems, 389 Butyl synthetic elastomers, 1466 applications in pharmaceutical sciences,
phosphate, 389 Butylated hydroxyanisole (BHA), 1554 393
properties of, 387 humectants, 996 isothermal titration, 1884
salts, types of, 2862 Butylated hydroxytoluene (BHT), 1554, 3348, scanning, 1884
selection of, 388 1708 Camag Automatic TLC Sampler III, 540
sodium phosphate, 836 humectants, 996 computer-controlled automatic developing
solution chamber, 541
neutralization titration, 3755 horizontal developing chamber, 541
preparation of, 1509 Cadila system, 1667 Camag linomat IV band applicator, 540
succinate, 1614 Caementum. See Cements. Cambric, oiled. See Silk, oiled.
Buffering agents Caenorhabditis elegans, 3147 Cambridge structural database (CSD)
use of, 388 Caffeine, 616, 2034 oxytetracycline in, 3956
water-soluble, 389 active ingredients, 4106 pharmaceuticals solids, 4105
Buffer-catalyzed reactions, 290 complexes of, 618 Camellia sinensis, 74, 2438
Buffering capacity, low rectal fluid, 1307 polymorphs of, 3436 10-Camphorsulfonic acid, 2159
Buginarium. See Bougies. Cahn–Ingold–Prelog sequence rule, CamposomesTM, 3579
Building monitoring and control system, 2144–2145 Camptothecin, biosynthesis of, 250
2881 Cake filtration, 3903 Canadian guideline, for analytical methods
Bulk drug substance (BDS), 2993 filtration method, 3886 validation, 94
Bulk powder properties, 3643 Calamine lotion, 995 Cancer
Bulk property detectors, 533 Calamitic mesophases cells
Bulk sampling, 2964 cholesteric, 1115–1116 hypoxic, 349
Bulking agents, 1822 columnar hexagonal, 1115–1116 drug treatment, 2469
crystalline, polymorphism in, 1823 nematic, 1115–1116 history of, 431
crystallization, conditions for, 1823 smectic, 1115–1116 human causes of, 431
freeze-drying cycle, 1275 Calcitonin, 2033, 2748 prostate, treatment of, 3360
general considerations, 1822–1823 biological effects of, 2707 treatment of, 1191
shelf life, 1275 intranasal delivery systems for, 1612 Cancer and burn injury, 3037
sublimation process, 1275 vaginal absorption of, 16 Cancer cell lines, 2444

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I18 Index

Cancer cells, endometrial, 71 Capsaicin, 2036 Capsules, gelatin, 2085

Cancer chemotherapy, 433, 1192, 1328, 2469 Capsular-shaped systems, 1293 CaptisolÕ, 1273, 1642, 3304
Cancer prevention Capsule fill capacity, 412 solubilizer, 1273
by diet modifications, 2443 dosing mechanism, 412 stabilizer, 1273
nutraceuticals in physical size, 412 Captopril, 1077
melatonin, 2444 type of formulation, 412 buccal formulation of, 1077
n-3-polyunsaturated fatty acids, 2444 Capsules, 3334 peroral (B) administration of, 1077
resveratrol, 2444 advantage of, 1291 pharmacodynamic parameters, 1074
soy isoflavones, 2443–2444 applications, 951 plasma concentrations, 1077
tea, 2444 coating, 414 sublingual administration of, 1077
by obesity reduction, 2443 combinations of, 412 Carbamazepine, 616, 845, 1019, 1244, 2862
Cancer risk assessment, 436 and its components, 992 cocrystals of, design of, 617
Cancer theory, multi-stage, 431 dissolution characteristics of soft gelatin, crystal growth of, 827
Candelilla wax, 4066 1867 crystal packing of, 617
superficial infections caused by, 3973 dry powder blends in, 406 monoclinic, 850
Candida albicans, 1033 elastic, 952 phases of, 2942
vaccine, 1356 empty, 408 photodegradation of, 620
Candida diphtheriae, 2479 standards for, 408 photodegradation products of, 623
Candy-base cookers storage for, 408 photomicrographs of, 630
high-speed atmospheric cookers, 2232 enteric-coated doses, 952 in polyethylene glycols (PEG), 769
types of, 2232 filling machine polymorph nucleation, 850
vacuum cookers, 2232 eauger, 409 solid dispersion of, 769
Candy-base manufacturing bench-scale, 410 solvates of, 617
candy-base properties, 2233 dry solid, 411 strategy of formation of cocrystals of,
final moisture content, 2233 instrumented, 411 621
heat-stable colors, 2233 formulation structure of, 618
manufacture of medicated lozenges, 2233 expert system, 417 trigonal, 850
precooking, 2233 hard gelatin, 1670 Carbamazepine cocrystals (nicotinamide or
Capillary cell membranes, 212 release, 415–417 saccharin)
Capillary columns, high resolution capability filling, 408 and solvates, molecular interactions in,
of, 516 capacity, 412 620
Capillary condensation, study of, 2590 dry solid, 411 stability of, 619
Capillary electrophoresis (CE), 98, 304, formulation types, 408 Carbamazepine dihydrate (CBZ(D)), faces of,
1189 liquid, 411 847
detection techniques, 2050 powder, 409 Carbamazepine nicotinamide (CBZ : NCT)
determination of compounds, 2050 from non-gelatin ingredients, 992 cocrystallization of, 629, 631
electrokinetic injection, 205 powder, 412 dissolution rates of, 620
electro-osmosis transport, 205 gelatin, 406, 952, 1616 melting points of, 619
electrophoretic separations of, 204 advantage, 407 solubility values of, 625
for immunoassays, 2050 hard, 992–993, 3335 Carbamazepine polymorphs
in body fluids, 204 soft, 3335 induction times of, 842
instruments, 204 softgel, 993 monoclinic, 842
migration rate of, 205 hypromellose, 406, 414 trigonal, 842
resolution separation technique, 204 for inhalation products, 415 Carbamazepine : saccharin (CBZ : SAC)
separation of macromolecules, 2050 in vitro testing, 413–414 cocrystallization of, 632
separation process, 205 in vivo testing, 414–415 dissolution rates of, 620
ultraviolet detection technique, 205 manufacture, 407 melting points of, 619
Z-shaped capillary flow cell, 205 gelatin, 407–408 Carbidopa–levodopa combination, example
Capillary electrophoretic immunoassay hypromellose, 408 of, 942
(CEIA), 1577 opaque, 666 Carbodiimide reagents, 1135
advantages of, 1577 pellets and granules in, 411 Carbohydrate and gastric homogeneity,
detection techniques, 1577 physical properties, 415 2868
resolving capabilities of, 1577 polymers, 406 Carbohydrate excipients
Capillary interaction, 35 sandimmune soft gelatin, 3349 lactose, 3304
Capillary isoelectric focusing (CIEF), 305 sealing, 412, 992 sucrose, 3304
Capillary rheometer, 1716 self-locking, 992 Carbohydrate residues, 1136
Capillary viscometers, 3141 shell manufacture properties oxidation of, 1136
Capillary zone electrophoresis (CZE) bloom strength, 992 Carbomer, 1881, 1882, 1887, 2407
charge=mass=ratios of, 206 viscosity, 992 molecular size of, 1887
detection sensitivity, 206 soft, 952, 992 properties of
non-chromatographic separation technique, starch, 952 mucoadhesive, 1887
206 structure for formulation of hard gelatin, Carbomer 934, 1887
Caprylic=capric acid, mono- and diglycerides 1673 Carbomer 934 alcoholic gel, 3265
of, 3348 Capsule weight data, 3508 Carbomer 940, 1887

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I19

Carbomer 941 gel, 3264 Carcinogenicity and mutagenicity, 2991 Cardiovascular drug propranolol, 320
Carbomer 971P (CarbopolÕ 971P), 2018 Carcinogenicity bioassays, 1416 Cardiovascular drugs, 1075
Carbomer gel, 1220 statistical analysis, 1417 captopril, 1077
Carbon black pigments, 1466, 1468, 1475 Carcinogenicity models, alternative, 440 glyceryl trinitrate (GTN), 1075–1076
Carbon dioxide, 2978 Carcinogenicity risk assessment, 436 isosorbide dinitrate, 1076
sources of, 2979 Carcinogenicity studies nifedipine, 1076–1077
Carbonless copy paper, 2316, 2330 FDA guidelines for, 689 Cardiovascular health
Carbonyl derivatives observation and measurement, 435 nutraceuticals treatment for
amine addition to, 2045 Carcinogenicity test, current, 433–440 coenzyme Q10, 2437
hydrolysis of, 2044 doses and route of administration, DHEA, 2437
Carbonyl groups, stretching vibration of, 433–434 lycopene, 2437
3415 duration, 434 melatonin, 2437
Carbopol 934, 1352 experimental conditions, 434 octacosanol, 2437–2438
Carbopol 934P, 1175 histopathological evaluation, 434–435 n-3-polyunsaturated fatty acids (PUFAs),
Carbopol gel, controlled release, 1356 historical control data, 435 2437
CarbopolÕ 971P, 2018 parameters examined, 434 pycnogenol, 2437
Carbosilane-poly(ethylene oxide) (CSi-PEO) statistical analysis, 435 resveratrol, 2437
dendrimers, 886 Carcinogenicity test legislation, history of, soy products, 2437–2438
Carboxamide homosynthon, 617 432 tea flavonoids, 2437–2438
Carboxy methyl cellulose, suspending agents, Carcinogenicity testing risk factors, 2437
1624 evolution in, 436 Caretek Medical, 1217
6-Carboxyfluorescein, permeation order, ICH topics on, 439 Carmustine (BCNU), from GliadelÕ, 186
1345 in future, 440–443 Carnauba wax, 765, 2010, 2018, 2330,
Carboxyhemoglobin, 358 Carcinogenic nitrosamines, 2991 4066
Carboxylate anion shifts, absorption band of, Carcinogenic potential, 2991 as coating material, 4072
701 Carcinogenic products, industrial use of, in cosmetic, 4066
Carboxylated styrene butadiene rubber latex 432 in food, 4066
foam, 1029 Carcinogens, 150 matrix, 767
Carboxylesterases genotoxic vs. non-genotoxic, 436–437 melting point of, 4074
A-esterases (arylesterases), 316 lipid peroxidation activation, 141 in pharmaceutical products, 4066
B-esterases, 316 non-genotoxic, 1413 polishing agent in sugar coating, 4066
C-esterases (acetylesterases), 316 positive control, 432 with valproic acid (VA), 768
esterase activity, 316 rodent liver, 1413 Carnivorous species, urinary reaction in,
hydrolytic reactions, 316 Carcinomas, 431 3962
non-specific enzymes, 316 lymphatic, 1610 Carotenoids, 143, 145, 2431
transport reactions, 316 Carboxypeptidases, 2718 autoxidation, 145
Carboxylic acid derivatives, 2041 Cardamon beneficial effects, 145
hydrolysis of, 2040 constituents, 1764 functions of, 143
Carboxylic ionophore antibiotic, prevention natural flavoring agents, 1764 CarpujectÕ, 1008
of coccidiosis, 3942 Cardiac arrest, treatment of, 349 Carrageenan, 1883–1884
Carboxymethyl-chitin hydrogel, 2031 Cardiac arrhythmias, 944 hydrogels of, 2029
Carboxymethyl cellulose, 1292, 1586, 1885 Cardiac drug (digoxin), intravenous injection Cartesian coordinates
Carboxymethylethylcellulose (CMEC), 41 solution formulation for, 1670 definition of, 714
Carboxyvinyl polymers, 2407 Cardiac dysrhythmias, 2639 molecular array, 714
Carcinoembryonic antigen, 1144 Cardiac glycosides, 245 Cartia XTÕ, 2255
Carcinoembryonic antigen (CEA) antibodies, Cardiac ischemia, 365 Cartilage tissue engineering, 2030
1149 Cardiac myocytes, cellular mechanisms=ion Cartridge-type depth filter, particle removal
Carcinogen, 1616 channels in, 2197 by, 4040
Carcinogenesis, 149 Cardiac Purkinje fibers, 2196 Carvedilol, 149
chemical Cardiomyopathy Case report forms (CRFs), 551, 2486
investigation of, 432 dilated, 1155 design process, 553
free radicals in, 139 ideopathic, 1156 Casein, 2381
solid-state, 432 Cardiopulmonary bypass (CEB), 361 Casson body
stages of, 149 surgery, 349 rheogram for, 3131
Carcinogenicity, 121, 689, 1413, 1616 circuit models, types of, 361 shear strain rate for, 3131
alternative models Cardiopulmonary resuscitation (CSR), Casson body model, 3132
evaluation 2632 shear thinning behavior, determination,
and implementation of, 441–442 CardioquinÕ, 1255 3132
use of, 442 Cardiovascular autonomic neuropathy, Cast stainless steels, 792
dosage selection, 1417 2872 alloys, 792
testing Cardiovascular disease, 588 heat-resistant, 792
history of, 431 drug biotransformation, in physiologic Castor wax, 4066
in past, 431–433 factor, 321 Catalase, 2034
weight-of-evidence approach, 442 Cardiovascular disease patients, 73 Cataplasm. See Plasters, medicated, 996.

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I20 Index

Catapres-TTSÕ, 984, 1087 CD4þT-cells Cefuroxime sodium

patches, 3362 activation, 3915 oily paste and oily suspension, 3959
scopolamine-releasing transdermal CD bands, 449 CelacolÕ, 995
therapeutic systems, 1087 CD detection Cell- and tissue-based electrodes, 1526
structural components, 1091 aminoacids, 456 Cell and tissue therapy, 265–266
Cataracting regime, 2355 carbohydrates, 456 Cell-based biosensors, 1525
Catechins, 147 of chiral substrates, 454 Cell-culture equipments, efficiency of, 1134
Catechol, 2142–2143 illuminating source of, 452 Cell deaths
Catecholamine, 213, 2148 light sources for, 450 apoptotic, 1159
autoxidation of, 149 lysergic acid diamide (LSD), 457 kinds of, 1159
enantiomeric forms in, 455 natural products in plant extracts, oncotic, 1159
Catecholestrogens, 148 456–457 programmed, 1159
catechol structure of, 148 proteins, 456 Cell dehydration, 161
Cathodic reductive stripping, 1499 sensitivity of, 453 Cell encapsulation, coacervation for, 608
Cathodic sputtering process steroids, 456 Cell growth, 1803
black coating, 3225 CD diode-array detectors, 453 regulation of, 1333
coating thickness, 3225 CD instruments, for analytical applications, Cell killing mechanisms, 1331
free electrons, 3225 451 Cell-mediated immune reaction, 46, 1421
principle of, 3219 CD spectra Cell membrane
Cathodic stripping voltammetry (CSV), for chiral complexes, 458 extrinsic proteins, 25
1498 for chiral Cu(ll)-L-tartrate metal complex, hydrophobic and electrostatic interactions
Cathodic voltage scan, application of, 1498 458 of, 25
Cation exchange chromatography, 303 in chloroform solution, 457 interdigitation of, 2680
Cationic compounds, determination of, for Cu(II)-D-histidine spectrum, 460 intrinsic proteins, 25
1512 for enantiomers, 454 polar molecules in, 25
Cationic dendrimers, 884 for mixed Cu(II)-peptide-D-histidine primary structure of, 25
inherent toxicity, 886 complexes, 461 structural model of, 25
for parenteral administration, 886 for mixed ligand complexes, 460–461 Cell membrane components, 1169
Cationic emulsifiers, 1552, 3261 for neuropeptide analyte, 461 Cell membrane glycoprotein, 1328
Cationic liposome-DNA complex (lipoplex), for polymixin and bacitracin antibiotics, Cell membrane lesion
1328 455 with cytoskeletal-antigen, 1159
Cationic liposomes for standard reference materials (SRM), with immunoliposomes, 1159
enzyme activity, 1165 454 ischemic process injury, 1159
of transfection efficiency, 1164 for underivatized partial racemic mixture, submicroscopic holes, 1159
Cationic surfactants, 826, 3586 459 Cell membranes, biochemical properties of,
Cattle for vitamin analyses, 455–456 212
ectoparasiticides classes, 3972 for water-soluble vitamins B2 and C, 455 Cell permeation enhancers, 32
gastrointestinal tristrongylids, 3948 CD spectropolarimeter double absorption enhancers, 32
half-life of phenylbutazone in, 3962 monochromator, 450 constituents of, 32
hydrophilic substances in, 3968 CD spectroscopic data, with quantitative drug absorption, 32
mineral deficiencies in, 3952 structure-activity relationships Cell-surface receptors, 1145
sebaceous glands, 3968 (QSAR), 461 Cell voltage change, 1511
skin, 3968 CDER. See Center for Drug Evaluation and Cellular drug permeability, 1302
treatment of frothy bloat in, 3950 Research. Cellular hyperplasia, 689
dimeticone emulsion, 3950 CDs. See Cyclodextrins. Cellular injury, 438
poloxaline, 3950 Cefadroxil, 1073 Cellular layers
Caveolae regions, clusters in, 1328 Cefamandole, distribution of, 579 contribution of, 3823
Cavitation bubbles, in stratum corneum, 3838 Cefazolin sodium Cellular macromolecules
Cavitation milling deflocculation behavior of, 4125 and cell sizes, 3821
cavitation erosion, 2344 flocculation behavior of, 4125 intercellular lipid matrix, 3821
energy dissipation, 2344 in non-aqueous media, 4125 intercellular space volume of, 3821
particle size reduction, 2344 zeta potential of, 4125 in epoxides, toxic effects, 313
CavitronÕ, 1642 Cefoperazone sodium in oxidation. of epoxides, 313
Cavity foam dressing, in wounds, 1029 oily paste and oily suspension, 3959 Cellular necrosis, 686
CBER. See Center for Biological Evaluation. CEFORMTM technology Cellular oncogenes, 645
Carbamazepine polymorphs monoclinic tablets manufactured with, 1111 drug delivery system, 1644
CD Cefoxitin, 1630 microcrystalline, 988
with absorbance detection, 453 chemical stability of, 88 Cellular protein expression profile, 3041,
with scanning double monochromator, Ceftazidime, 390 3042
450 CeftinÕ, 1256 Cellular receptors, 1327
CD-1959, absorbance detector of, 453 Ceftiofur sodium, 3955 Cellular vesicular processes, 2723
CD-active absorption bands, 457 Ceftriaxone Cellulose, 2159
CD-active analytes, multiple overlapping free fraction of, 3036 absorption and desorption, 2375
bands from, 454 interaction of, 3032 change in bulk solid properties for, 2380

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I21

[Cellulose] [Center for Drug Evaluation and Research Cervical mucus, 1341
chiral recognition properties of, 2160 (CDER)] Cervicovaginal cancers, 1353
chromatographic procedures for, 2160 good manufacturing practices (cGMPs), oxytetracycline in, 3956
derivatives, 2160 1783 Cetacaine, 901
differential scanning calorimetry of, 2377 guidelines, 1402 Cetyl=stearyl alcohol, 1127
enantioselective properties of, 2159–2160 NDA, 1783 Cetyl=stearyl sulfate, 1127
force vs. hardness for, 2380 Center for Drug Evaluation and Research Cetyl dimethylammonium bromide, 901
Karl Fisher titrations for, 2377 Office of Compliance (CDER-OC), Cetylmethylammonium bromide, 2674
microcrystalline, 1613, 1614 3189 Cetylpyridinium chloride, 2672
moisture effect in, 2380 Centers for Disease Control and Prevention Cetylpyridium chloride, 897
monolayer capacities of, 2380 (CDC), 1044 Cetyltrimethylammonium bromide (CTAB),
pharmaceuticals applications, 2160 Center for Veterinary Medicine (CVM), on nanoparticle surface, 1193
polymer chains of anionic, 1734 1401 Charge-coupled devices (CCD),
retention and enantioselectivity of, 2160 Central and Eastern European countries 3400
sorption isotherms for, 2373 (CEEC), 1597 array detectors, 3474
tablets, 2380 pharmaceutical trade with, 1597 chips, 3400
thermal gravimetric analysis of, 2376 Central nervous system (CNS), 946 for chromatograms, 542
water in, 2380 depressants, 1041 cooling of, 3400
Cellulose acetate butyrate (CAB), 2012 drug penetration into, 2635 in spectrophotometer, 3464
Cellulose acetate phthalate, 420, 606, regeneration, 2029 Charge-transfer complexes, in organic
829, 992 stimulants, 1043–1044 molecules, 3471
Cellulose-based polymers, 2012 tissue replacement in, 2029 Charged particle activation analysis (CPAA),
Cellulose derivatives, 994, 1885, 2407 Central nervous system (CNS) ischemia, 139 3091
cellulose fiber, 3231 Central Pharmaceutical Affairs Council Charged particulate radiations, 3541
hydroxyethylcellulose (NatrasolÕ 250), (CPAC), 2771, 2836 Charta amylacea, 953
995 Central venous catheters (CVC), types of, Charta cerata. See Cachets.
methylcellulose (CelacolÕ), 995 1003 Charybdis technologies, solid-phase synthesis
microcrystalline cellulose, 995 Centrifugal elutriator, 2590 applications, 3006
microscopic images, 3231 Centrifugal force, application of, 1248 Chatelier’s principle, 3004
sodiumcarboxymethylcellulose, 995 Centrifugal sedimentation, 3888 Chatillon digital force gauge, 2017
super disintegrants, 3231 in liquid–liquid mixtures, 3888 Cheesegrater effect, 911
Cellulose ether, 1661, 1886 in solid–liquid mixtures, 3888 Chelates, properties of, 3759
Cellulose membrane filter (CMF), 2310 types, 3889 Chelating agents, 1612, 2862
Cellulose nitrate, dissolved in volatile Centrifuging regime, 2355 amino acids, 1625
solvent, 3901 CepacolÕ, 897 citric acid, 1625
Cellulose sponge, 1027 Cephalexin, 1021 effect of, 2862
Celluloses Cephalosporin antibiotics, 84, 256, 1229 excipients, 1625
ball milling studies, 4060 Cephalosporium acremonium, 256 in parenteral products, 1625
Brunauer, Emmett, and Teller (BET) Cerates, unctuous substances, 953 tartaric acid, 1625
model, 4059 Ceratum. See Cerates. Chelatotherapy, significance of, 697
glass transition temperatures, 4061 Cerebral blood flow (CBF), 364 Chemical, excipients for, 1638
isotherm analyses of, 4059–4060 Cerebral hemodynamics, influence on, 361 Chemical absorption enhancers, 13
Cellulosic dressing products Cerebral ischemia, 363 Chemical Abstracts Service (CAS) number,
absorbent cotton, 1031 Cerebral spinal fluid, 1626, 2991, 3054, 1949
gauze, 1031 3059 Chemical additives
Cements, sterilizing effect, 953 administration, endotoxin limit for, 3059 container closure system component,
Center for Biologics Evaluation and Research Cerebral vascular accidents, 944 1695
(CBER), 592, 1696, 1779, 1941 Cerebral vascular disease, 72 with elastomer=polymer matrix, 1694
for biological products regulation, Cerebrovascular function, 241 Chemical adsorption, 34
57, 59 CerebyxÕ, 2250 Chemical amplification, 1526
Center for Devices and Radiological Health product labeling, 2251 Chemical and aggregation stability, of hGH,
(CDRH), 1401 CerebyxÕ, 1255 1828
Center for Drug Evaluation and Research Cerecloths, 953 Chemical antimicrobial agents, 2983
(CDER), 1696, 1779, 1941, 2774 Cereolus, urethral bougie, 953 See Bouiges. Chemical carcinogenesis
abbreviated new drug applications Cerium(IV) classification of, 436, 437
(ANDAs), 1783 oxidizing reagent, 3758 investigation of, 432
drug master file types, 1403 Cerumen Chemical decomposition, solid state reaction,
type 1, 1403 applications of, 2477 4112
type 2, 1403 chemical composition of, 2475 Chemical degradation, 1827
type 3, 1403 composition, 2477 Chemical degradation, of hGH, 1828
type 4, 1403 constituents of, 2475 Chemical disinfecting agents
type 5, 1403 preparations, 2476 categories of, 2208
good clinical practices (GCPs), 1783 replacement products for, 2477 non-oxidizing, 2208
good laboratory practices (GLPs), 1783 self-cleaning mechanism, 2477 oxidizing, 2208

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I22 Index

[Chemical disinfecting agents] Chemiluminescence, 1576, 3387, 3401 Chiral HPLC system, 453
properties of definition of, 151 Chiral materials, 445, 446, 453
non-irritating, 2208 detection system, 1576 Chiral mobile-phase additives, 2157
non-sensitizing, 2208 in living organisms, 3401 Chiral stationary phases (CSP)
Chemical disinfection reactions, 3401 components, 2159
antimicrobial compounds, 2206 Chemiluminescence immunoassay in liquid chromatography, 2159
disadvantages of, 2206 automated immunoassay analyzers, 204 structural features
FDA guidelines, 2206 bioluminescent reactions, 2057 aromatic p-electron systems, 2159
of soft contact lenses, 2206 chemical modification, 2057 cellulose, 2159–2160
testing process, 2206 chemiluminescent compounds, 2057 crown ethers, 2159
Chemical enhancers, enhancement of, 2931 chemiluminophores, 204 cyclodextrins, 2159
Chemical equilibrium, oxidation–reduction definition, 2057 metal complexes, 2159
titration, 3757 dioxetanes, 2058 proteins, 2159–2160
Chemical gels, 3264 direct, 2058 starch, 2159–2160
Chemical hardening process, 606 enzyme-mediated, 2059 synthetic polymers, 2159–2160
Chemical hydrolysis, 1136 exergonic reactions, 2057 Chiral substances, enantiomeric purity in,
Chemical ionization luminescence efficiency, 204 459
reagent gas, 1701 in oxidation products, 204 Chiral substrates, 454–455
spectrum for, 1702 oxidation–reduction reactions, 2057 Chirality, 2143
thermodynamically controlled process, sensitivity, 204 off ephedrine, 2145
1702 Chemistry, Manufacturing and Control Chiroptic detection, 452
Chemical methodologies, for reduction of (CMC) committee, 3930 CW lasers in, 452
barrier function, 1311 Chemokine receptor, characterstics of, 3125 dye lasers in, 452
Chemical modification, types of, 2727 Chemometric tools, 3383 Chiroptical analytical methods, 445
Chemical modification approach, use of, application of mathematical methods, Chiroptical detection
2713 3630 direct, 453–454
Chemical penetration enhancers application of statistical methods, 3630 for testosterone and dihydrotestosterone
penetration enhancing effects, 3846 artificial intelligence algorithms, 3383 analysis, 451
solubilizing enhancing effects, 3846 cluster analysis, 3383 in chromatography, 452–453
for transdermal transport, 3846 definition, 3630 wavelength ranges for, 451
use of, 2741 development of NIRS methods, 3630 Chiroptical dispersion spectra, 447
Chemical process industries (CPI), 782 factor analysis, 3383 Chiroptical methods
Chemical radiation, mechanism of, 3542 in interpretation of analytical data, 3630 analyte selection in, 451–452
Chemical reactivity, 714 pattern recognition, 3383 applications of, 454
Chemical sanitization, 4047 Chemoreceptor trigger zone, 72 circular dichroism (CD), 445
Chemical shift anisotropy (CSA), 3298, Chemotherapeutic agents, 3008 optical rotatory dispersion (ORD), 445
3456 cytotoxic action of, 1138 polarimetry, 445
Chemical shifts Chemotherapeutic and immunological instrumentation for, 450–451
definition, 3441 agents, 1145 detector, 450
external magnetic field, 3441 Chewable tablets, 1610, 1769 polarizing elements, 450
molecule shielding, 3441 advantages, 991 sample holder, 450
nuclear shielding effects, 3441 Chi-square distribution, 3491 stable unpolarized illuminating source,
spectral dispersion, 3442 in goodness-of-fit test, 3488, 3491 450
Chemical skin permeation enhancers, 2743 Chi-square statistics, 3486 wavelength-selection device, 450
Chemical stability Child Health and Human Development Chitin, 160
homogeneous drug systems, 2222 (NICHD), national institute of, Chitosan
pH-solubility profiles, 2222 2630 absorption-enhancing effect, 2686
pH-stability profiles, 2222 Chimeric antibody, construction of, 1133 application, 2030, 2686
Chemical substances Chinese hamster ovary cells, 1414 capsule manufacture, 406
inventory of, 1948 Chinese Pharmacopeia Commission (ChPC), deacetylation process, 2686
monographs on, 2829 2856 degradable, 1643
International Conference on Chinese restaurant syndrome, 1616 hydrogels of, 2029
Harmonisation (ICH), 2829 Chiral amino alcohols, 2159 mucociliary transport system, 2686
safety of, 1948 Chiral chromatographic methods, N-acetyl-glucosamine, 2686
Chemical transdermal enhancers problems in, 453 nasal administration, toxicity by, 2686
chemical permeation enhancers, 3846 Chiral chromophores, in near-UV, 455 nasal peptide absorptionm, effects on,
drug absorption, 3846 Chiral derivatizing agents, 453 2685
mechanisms of, 3846 Chiral drugs polymers, 3921
permeability coefficient, 3846 enantiomeric purities of, 453 polysaccharide, types of, 2686
incompatibility, 3846 quality control (QC) for, 459 preparation of vaccine formulations, 3921
optimal patch system, 3846 Chiral excipients processing, 2029–2030
transdermal flux enhancement, 3846 benzocaine, 455 salmon calcitonin, 2686
transdermal transport, 3844 lidocaine, 455 Chitosan–acetic acid solutions, 2029–2030
Chemical vapor deposition (CVP), 3793 procaine, 455 Chitosan-based dosage forms, 999

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I23

Chitosan-based transdermal drug-delivery [Chlorpheniramine maleate] [Chromatographic systems]

systems, 2036 wax matrix development of, 196
Chitosan-coated formulation, 1177 by spray-congealing agglomeration evalution of, 2849
Chitosan-coated liposomes process, 769 gel permeation, 532
adhesion properties, 1179 Chlorpheniramine maleate (CPM)-loaded hydrophobic interaction, 531
constituents of, 1179 non-pareils, coating of, 766 ion-pair, 530
Chitosan delivery systems, 2035 Chlorprothixene, 2146 mobile phase, 196
Chitosan gel cores, 1236 Chlorofluorocarbon (CFC) propellants classification of, 196
Chitosan microspheres, 1236, 2035 ozone-depleting, 1285 modes of, 529
Chitosan nanoparticles, 1186 Chlorthalidone dispersion, prepared by normal phase, 528–529
Chitosan salts, 17 spray–drying process, 775 reverse phase, 529–530
Chlamydia infection, 1221 Chocolate flavor, 1770 sample preparation in, 537
Chlorambucil antimelanoma globulin, Cholecystokinin, production of, 689 silica surface in, 529
studies with, 1140 Cholera toxin, 2746 size exclusion, 532
Chloramphenicol, 943, 1019, 2862 Cholesteric liquid crystals stationary phase, 196
Chloramphenicol derivativesm, veterinary color play of, 1115 classification of, 196
drugs, 3941 in cosmetics, 1130 paper chromatography, 196
Chloramphenicol palmitate Cholesterics, 1115–1116 thin-layer chromatography, 196
blood concentration of, 3313 Cholesteryl esters, birefringence of, 1115 Chromatography, reversed-phase
solubility of, 3313 Cholestral sulphate, accumulation of, 3822 guideline for validation of, 94
Chloramphenicol stearate and colloidal silica, Cholestyramine Chromatography data system, as hybrid
interaction between, 1613 and Colestipol, 1397 system, 6, 2165
Chlordiazepoxide, distribution of, 1018 administration of, 1397 Chromatography systems and laboratory
Chlordiazepoxide (LibriumÕ), 1041 application of, 1397 automation, 740
Chlorhexidine, 891, 896 in gastrointestinal (GI) tract, 1397 Chromium carbide precipitation, 793
delivery system, 903 Cholestyramine (Questran), 1397 Chromium-labeled chicken red blood cells,
Chlorhexidine gluconate (HibiclensÕ), 896, Cholestyramine resin, bioequivalence testing, 1421
904, 2031, 2478 172 Chromium-labeled target cells, 1421
iodine-allergic patient, 2478 Choline oxidase, 2030 Chromobacterium violaceum, 256
spectrum of antimicrobial activity, 2478 Choline salicylate, preparation of, 3003 Chromogenic reactions, endpoint of, 3059
for surgical prophylaxis, 2478 Cholinergic toxicity, 76 Chromophores, 447
Chloride–bicarbonate exchangers, 27 Chondroitin, 73, 2431 Chromosome aberrations, 437, 689
Chloride ion-containing resins, 2124 active constituents in, 2446 Chromosomal damage, 1413
Chloride ion management, 2123–2124 applications, 2437 Chromosome damage tests, 120
Chlorinated-fluorocarbons, 2093 clinical trials, 2437 Chrondocyctes, 266
Chlorobutyl rubber, elastomer, 1466 effects, 2436 Chronic active hepatitis, 1019
Chlorocresol, preservatives of excipient, and glucosamine combination, 2446 Chronic alcoholism, 2871
1626 incidence, 2437 treatment of, 703
Chlorofluorocarbon (CFC), 1541, 1617, Chondroitin sulfate, matrix systems of, Chronic bioassay, 443
1697 1237 deficiency of standard, 438
based pMDIs, 2107 Chromamycin A3, 936–937 Chronic disease, treatment of, 2741, 2752
based suspension formulations, 2273 Chromatograms, documentation of, 542 Chronic hemolysis, 365
consumption, 2270 Chromatographic analysis, requirements, Chronic inflammatory process, 116
in metered dose inhalers (MDIs), 2270 3028 Chronic irritation=mitogenic activity, 443
in pressurized metered dose inhalers, Chromatographic assays, 1572 Chronic liver disease, 586
2101 implications for validation, 1572 Chronic myeloid leukaemia, treatment of,
physical properties of, 2101, 2107 Chromatographic characterization, 1326
propellants, 2077, 2101, 2269 accumulation of, 2853 Chronic obstructive pulmonary disease
Raoult’s law to, 2270 Chromatographic enantioseparations, 2156 (COPD), 2093
surfactants in Chromatographic impurity, determination of, Chronic renal failure patients, 363
lecithin, 2273 94 Chronic stable liver disease, propanolol in,
oleic acid, 2273 Chromatographic methods, 3 3030
sorbitan trioleate, 2273 application of, 3 Chronic toxicity studies, 124
types, 2270 characterstics of, 2852 Chronic toxicity testing, 443
use of, 1061 extrapolation of, 3 Chronic toxicology, 118
Chloroprene, 2275 HPLC (high-performance liquid carcinogenicity
Chloroquine, 2862 chromatography), 3 studies, 1950
Chloroquine diphosphate, 2941 Chromatographic peak purity, investigation Chronogest vaginal pessary, 1351
Chlorpheniramine, 2416 of, 99 ChronomerTM, 186
Chlorpheniramine maleate, 768, 2008, 2016 Chromatographic purity, of analyte, 3471 ChronomerTM. See also AlzamerÕ.
influence of drug loading on, 2016, 2018 Chromatographic separations, types of, 303 ChronotopicÕ system, 1255, 1288
PEG 3350 influence on, 2016 Chromatographic systems, 196 ChronsetÕ system, 1294
release from extruded matrix tablets, 1294, affinity, 532, 1134 Chute splitting, 2966
2016 chiroptical detection in, 452–453 Chymotrypsin inhibitor, 2726

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I24 Index

Chymotrypsinogen, 2718 Clarithromycin (Biaxin), 1399 Clinical data management systems, 551
Ciclopiroxolamine, 1353 ClaritinÕ, 1256 advantages of, 551
Ciclosidomine, acid=base hydrolysis, 390 as allergy releiver, 2422 analysis and reporting, 558
Ciliotoxic effect, 15 chronic hives treatment by, 2422 derivation, 558
Cimetidine, 1398, 2445 FDA application to, 2422 electronic submission of CRFs, 558
histamine H2-receptor antagonists, 1399 switch, 2422 extracting data, 558
metabolic pathways, 1398 Clark techniques, 1971 tools, 558
sedative effect of, 1398 Class 100 non-laminar airflow, 2175 study conduct, 555–558
Cimicifuga racemosa (Black cohosh), 69 Classic calomel electrodes, disadvantage of, data entry, 555
Ciphergen’s protein chips, 2198 1505 database closure, 556
Cipro otic drops, 2481 Classic d.c. polarography, 1494 discrepancy management, 556
Ciprofloxacin, 1190 Classic polarography, 1492 electronic audit trail, 556
delivery of, 185 Classic receptor-occupancy theory, 2802 laboratory data, 557
Ciprofloxacin (Cipro), 1397 Classical Clapeyron equation, 2935 ongoing monitoring, 556
Circadian cycles, 3823 Classical coupling methods, classification of, pharmacokinetic data, 557
circadian rhythmicity, 3823 1136 receipt of data, 555
Circadian rhythm, 1287, 2868 Classical light scattering, use of, 1056 study performance metrics, 557
body function on, 1287 Clathrin-coated vesicles, 2724 thesaurus management systems, 557
Circular demountable cell, for liquid sample Clathrin, definition of, 2724 study set-up, 551–555
analysis, 3413 ClaversalÕ, 1254 CRF development, 553–554
Circular dichroism, 448–449 Clay minerals, 637 data quality specifications, 554–555
advantage of, 448, 451 Clean-in-place (CIP), 748, 2882 protocol development, 551
for analysis cleaning equipment methods, randomization, 554
absorbance methods, 452 1581 standardization, 551
of bulk samples, 449 for machinery, 381 Clinical development, phases of, 2806,
and circularly polarized luminescence measurement4 3021
(CPL), 452 practices, 381 Clinical drug development, stages in, 563
and fluorescence, 452 specific equipment design, 381 Clinical drug research, 563
Xe arc lamps in, 450 spray balls, 1583 Clinical research
biological extracts spray devices, 1583 development process, 1925
direct analysis of, 455 dynamic, 1583 GCP regulations, 1925
elliptically polarized light in, 449 riboflavin test, 1583 physician investigators, 1925
for detection sanitary, 1583 requirements of FDA, 1925
of alkaloids, 455 validation of, 1583 Clinical Research Associates (CRAs), 2486,
of aromatic amines, 455 validation, 381 3071
of beta-lactam antibiotics, 455 Clean-out-of-place (COP), cleaning Clinical research project, 562
of morphine alkaloids, 455 equipment methods, 1581 Clinical studies
of vitamins, 455 Clean-zone concept, 1944 biological products, 1784
method of, 703 efficacy, 381 human drug products, 1784
Circular dichroism (CD) detection=ligand Cleaning agents new drug applications (NDA), 1784
exchange aqueous versus non-aqueous, 1583 Clinical supplies inventory system, 733
for assays cleaning equipment process, 1583, Clinical supplies labeling, 733
of oligopeptides, 459–460 1880 Clinical supplies manufacturing, 2884
of peptides, 459–460 concentration, 1584, 1585 Clinical supply quantity requirements,
Circular dichroism (CD) spectroscopy, 1879 cleaning solution, 1584 2877
Cirrus nebulizer, 2096 water quality, 1584 Clinical trials, management of, 559, 735
Cis–trans isomerization, 2860, 2942 Cleaning parameters CLOGP method, fragmental constant, 8
Cisplatin cleaning agent concentration, 1585 Clomethizole edisilate, 3347
anticancer agent, 885 equipment cleaning process, 1584 Clonazepam, 2862
liposome formulation of, 1332 impingement, 1585 Cloned human receptors, 3118
toxic side effects, 885 mixing, 1585 baculovirus system, use of, 3119
water solubility, 885 temperature, 1584, 1585 in vitro mutagenesis, use of, 3119
Cisplatin suppositories, 1353 prewash, 1585 mammalian cells, 3119
Citrate, buffer salt, 1623, 2862 rinse, 1585 pharmacological screening method, 3118
Citrate esters, 2008 time, 1584 receptor density, 3119
Citric acid, 1355, 1610 aspects, 1584 receptor function, analysis of, 3119
Citrinin, biosynthesis of, 230 water quality, 1585 receptor phosphorylation, 3119
Citrus aurantium, 1765 Cleaning standard operating procedure receptor structure–function relationships,
Citrus essential oils, liquid flavors, 1764 (SOP), 1581, 1591 3119
Citrus limonum, 1764 Cleaning validation guidelines, of drug recombinant expression systems, 3118
Citrus sinensis, 1765 products, 1580 X-ray crystallography, 3119
Clarithromycin Clenbuterol and salbutamol residues, Closed-cycle refrigerators, construction of,
bitter taste masking of, 769 in animal urine, 545 1052
polymorphism in, 3302 Clindamycin, 1352 Clostridium sporogenes, 327

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I25

Closure–drug interactions, types of, 1477 [Cocrystal(s)] [Collaborative document types]

Clotrimazole, 2478 dissociation, equilibrium equations, program files, 2556
Cloxacillin, slow-release intramammary 627 SAS logs, 2556
preparations, 3959 dissolution rates of, 620–621 Collagen, 1882–1883
Clysters. See Enemas. diversity of, 615 asymmetrical molecular structure of, 1882
CNS (central nervous system), 48 formation, 620, 621 drug delivery system, 1644
depressants, 49 by cogrinding crystalline components, gelatin material from, 406
additive effect, 1394 631 implants (Lacrimedics), 1883
alcoholic beverages, 1394 principles of, 617 in hydrogel preparation, 190
barbiturates, 49 processes and mechanisms of, 628–633 Collagen disks, applications, 1223
benzodiazepines, 49 reaction for, 632 Collagen dressings, 1035
malnutrition, 1394 by solid-state mediated processes, Collagen shields
side effects, 1394 631–633 drug delivery of, 1222
in humans, 812 solution-mediated processes, 628–631 amphotericin, 1223
in mice, 812 solvo-thermal methods, 628 chlorhexidine gluconate, 1223
systemic effects on, 812 growth of, 629 erythromycin, 1223
CNS drugs. See Psychoactive drugs. hydrate formation, 619 povidone-iodine, 1223
CNS-effective sedative hypnotics, 1043 hydrogen bond patterns in, 617 procaine penicillin, 1223
Coacervation melting point, 619 silver nitrate, 1223
and microencapsulation, 600 pharmaceutical properties of, 615 tobramycin, 1223
classification of, 600 properties of, 619–628 technology, 1223
commercial application of, 600 solubility, 621–622 types
definition of, 600 as function of ligand concentration, Bio-Cor, 1222
taste masking of drug, 1106, 1107 621–622 ProShield, 1222
type of, 607 as function of pH, 626–628 Soft Shield, 1222
Coacervation dispersion, composition of, solvates, 615 Collagens (gelatin), mammalian, 1612
603 Cocrystallization Collette and Fielder mixers, 3194
Coagulation process, 4122 by mixing solutions of reactants, 629 Colligative properties
Coal-tar colors, 648 pathways, 632 boiling point elevation, 3770
Coal-tar hair dyes, 800 rate of, 632 definition, 3773
Coalescence reaction, 630, 631 degree of dissociation, 3773
continuous phase, 1555 Cocrystals and solvates, examples of, 616 of dilute solutions, 3770, 3773
liquid-bridging state, 2259 Code of Federal Regulations (CFR), 1658, of electrolytes, 3773, 3779
liquid saturation state, 2259 1685, 1781, 2237 freezing-point depression, 3770
mechanism, 36, 1555 relevant sections of, 1686 osmotic pressure, 3770
of melt agglomerates, 2259 Codeine, 1040 Raoult’s law, 3770
random collisions, 2259 Codex Alimentarius, 3991 van’t Hoff factor, 3774
size-enlargement process, 2259 Codex alimentarius commission, 3991 vapor pressure lowering, 3770
Coarse suspension, 3597, 4125 Coefficient of variation (CV), 2765 Collision-induced decay (CID), 3046
Coated pits, definition of, 2724 Coenzyme Q10 Collodions, 953
Coated tablets, 2408 and K-vitamins interactions, 2445 Collodium. See Collodions.
Coating layer, mechanical properties of, and warfarin interactions, 2445 Colloid drug-carrier systems
1291 effects, 2439 types of, 642
Coating pills, 953 Cohesive blends Colloid drug-delivery system, 636, 642
gelatin, 953 electrostatic force, 3204 Colloid mills, 995–996, 3268–3269
sugar, 953 types of cohesive forces, 3204 Colloid oncotic pressure, 356
Coating processes, of congealable material, van der Waals force, 3204 Colloid osmometers, 3776
765–766 ColazideÕ, 1254 Colloidal aggregates, internal structure of,
Coating system, devolopment of, 744 Colchicines, biosynthesis of, 244 1052
Coca-ColaÕ, 1043 Cold denaturation, 1810 Colloidal and polymeric drug delivery
Coca leaf-chewing, 1043–1044 Cold vapor technique, for mercury, systems, 1049
Cocaine 3368 Colloidal carrier systems
abuse, 1044 Colestipol (Colestid), 1397 types of, 643
biosynthesis of, 248 Colgate tartar control dentifrice, 899 use of, 2743
forms, 1044 Colgate tartar control mouthrinse, 900 Colloidal dispersions
speedball of, 1044 Colgate TotalÕ, 896 Brownian motion, 2386
Cocarcinogens, 1425 Colistin, 2479 particle aggregation, 2386
Cochan–Armitage test, 433, 435 Collaborative Agreement of Drug Regulatory Colloidal drug carriers, 1191
Cockroft–Gault equation, 1907 Authorities of European Union Colloidal drug delivery system, study of, 1054,
Cocoa butter base, 996 Associated Countries (CADREAC), 1195
Cocrystal(s) 1597 Colloidal gold staining, 1173
carbamazepine, design of, 617 Collaborative document types Colloidal particles
chemical stability, 620 datasets, 2556 Stokes settling radius of, 639
components, 616–619 graphics, 2556 surface of, 1052

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I26 Index

Colloidal plasma volume, 358 [Colon] Colon-specific drug delivery systems

Colloidal silica, 1613 drug permeability in, 1229–1230 (CSDDS), 1254
and chloramphenicol stearate Enterobacteriaceae, 1230 Colon-targeted delivery capsule
interaction between, 1613 factors affecting drug absorption from, components, 1234
desiccant effect of, 42 1228–1229 design
in tablet formulation, 3659 functions acid-soluble layer, 1234
Colloidal silicon dioxide, glidant, 3661 in colonic microorganisms growth, 1228 active ingredient, 1234
Colloidal suspension, 3597 in drug absorption and secretion, 1228 enteric layer, 1234
in liquid medium, 4126 in expulsion process, 1228 gelatin capsule, 1234
Colloidal system as storage reservoir, 1228 hydrophilic layer, 1234
electric double-layer properties, 4121 hydrolytic enzymes of organic acid, 1234
interaction energy curve, 4121–4122 a-L-arabinosidase, 1231 pH sensitivity of, 1234
physical stability, 4121 b-galactosidase, 1231 pressure-controlled, 1235
determination, 4121 b-glucuronidase, 1231 time-release principles of, 1234
electrostatic repulsion, 4127 b-xylosidase, 1231 Colony-forming units (CFU), 1222
Colloidons, 994 polysaccharides delivery to, 1235 Colony forming units (CFUs), 2303
Colloids, 2989 principal role of, 2870 Colony-stimulating factors (CSFs), 270
anionic preservatives rectosigmoid, 1228 Color
interaction of, 2989 reductive enzymes of coal-tar, 648
benzalkonium chloride, 2989 azoreductase, 1231 designations, 649
classification of, 636 deaminase, 1231 discovery of, 648
definition of, 636 nitroreductase, 1231 in distinguishing drugs, 648
non-ionic surfactants, 2989 urea dehydroxylase, 1231 in elimination of medical errors, 648
particle size measurement, method of, 639 spastic, 1228 label requirements of, 649–650
transmission electron microscopy (TEM), transversale, 1228 lakes, 649
639 Colon and lung cancer models, 1139 in pharmaceuticals, 648
phenylmercuric acetate, 2989 Colon cancer, 438 for taste masking, 648
polyethylene glycols, 2989 Colon cell lines, 1302 safety measures of, 649
property of, 638 Colon delivery, 2727 sources
scanning electron microscopy (SEM), 639 Colonic bacteria, 1615, 2872 animal, 648
tragacanth and acacia, 2989 Colonic bacteria, action of, 1256 minerals, 648
concentration of, 2989 Colonic diseases plant, 648
ultrasound vibration potential (UVP) colorectal cancer, 1228 synthetic, 648
effects in, 4119 Crohn’s disease, 1228 Color Additive Amendment, 432, 651
uses of, 636 spastic colon, 1228 Color Additive Petitions, 660
versus emulsion, 636 treatment of, 1254 Color additives, 799
Collunarium. See Solutions, nasal. Colonic drug absorption, 1303 certified, 800
Collutorium. See Washes, mouth. Colonic drug absorption enhancers classification of, 800
Collyria, 1220 bile salts, 1231 dyes and pigments, 799
Collyrium. See Solutions, ophthalmic. chelating agents, 1231 non-certified, 800
Colon fatty acids, 1231 Colorants, 1734
anaerobic bacteria in mixed micelles, 1231–1232 application criteria, 664–665
Bacteriods, 1230 non-steroidal antiinflammatory agents, approved in food use, 660
Bifidobacterium, 1230 1231 in Canada, 663
Eubacterium, 1230 surfactants, 1231 in Europe, 661–662
ascendens, 1228 Colonic drug delivery in Japan, 664
azo-based polymeric delivery systems of, Colonic homogenates, 2727 BIRYO limit for, 660
1238 Colonic microflora, enzymatic activity of, categories of synthetic colors for
bacterial flora in, 1229, 1231 1254 drugs and cosmetics (D&C), 651
bacterial growth regulation in, 1230 Colonic mucosa, 2727 externally applied drugs and cosmetics
cecum, 1228 pharmacological action at, 1256 (external D&C), 651
characteristics of Colonic transit time, 2872 foods, drugs, and cosmetics (FD&C),
anatomical, 1228 Colon-specific drug delivery 651
biochemical, 1228–1229 pH and transit time of, 1229 definition, 651
physiological, 1228 physiological factors of, 1228 drug, 648
characteristics of drugs favoring, 1231 polysaccharides for exempt from U.S. certification, 657–659
Clostridium enterococci, 1230 amylose, 1235 applications, 656
definition of, 2713 chrondroitin sulfate, 1235 b-carotene, 656
descendens, 1228 dexamethasone in, 1236 organic and inorganic colorants, 656
drug absorption inulin, 1235 sources, 656
barriers of, 1229–1230 pectin and its salts, 1235 in foods, drugs, and cosmetics, 648–649
enhancers, 1231–1232 strategies, 1232 history, 648
drug delivery, 1228 polymer-based approach, 1233 multinational pharmaceutical applications,
drug metabolism in, 1230–1231 prodrug-based approach, 1232–1233 660–664

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I27

[Colorants] Common Technical Document (CTD), Compressed tablet lozenges

non-harmonized regulations of, 650–651 acceptance of, 4102 artificial sweeteners, 2234
permanent listed, 651–653 Compaction, adsorption in, 37 batch release testing, 2235
properties and applications of, 668–669 Compaction behavior, mathematical components, 2233
provisionally listed, 654–655 constants, 3612 direct compaction, 2234
regulatory status of, 800 Compaction process, roll speed for, 3166 dry granulation, 2234
safety of food, 649 Compaction theory, 3160–3164 formulation factors, 2234
U.S. certified, 652–655 density–pressure curves, 3612 physical attributes, 2234
Color Index (CI) numbers, 651 Compactor machine design, operating preparation method, 2233
Color lakes, definition of, 800 parameters of, 3172 in process testing, 2234, 2235
Colorectal cancer, 1228 Compactor roll blocks, 3167 slow dissolution, 2235
Colored complexes, titration of, 3760 Compactor roll configuration, 3166 stability, 2235
Coloring agents, 648 designs, 3167 wet granulation, 2234
color shades, 2225 Compactor rolls Compressed tablets. See Tablets.
colorants in medicinal products, 2225 in horizontal plane, 3162 Compression calorimeter, 403
dyes, 2225 nip angle, 3163 Compression feed screw, and fixed roll
psychological effects, 2225 powder regions, 3163 system, 3167
Column chromatography, 3004 Compartmental pharmacokinetic models, Compression force transducer, tablet press
column packing 2759 instrumentation, 3689
constituents of, 197 Compendial standards Compression test
liquid phase of, 197 principles of, 2848 advantage of, 1737
Columnar cells technical features of, 2847 of coated, 1737–1738
apical surface of, 2680 Compendial test, development of, 3056 enteric-coated soft gelatin capsules, 1737
types of, 2680 Compendial water, definition of, 4039 Compressional bonding
Columnar epithelial cells, 577 Compendial water systems theories
single layer of, 1246 compendial water systems, 2881 intermolecular, 3161
Combinatorial chemistry, 265 design of, 2880 liquid-surface film, 3161
chemical diversity, 1364 Competitive assays, 2048 mechanical, 3161
chemical information systems, 1365 enzyme-labeled antibody, 2053 CompritolÕ, 1258
methodology, 1364 enzyme-labeled antigen conjugate, 2052 Compton scattering, 3541
organic synthesis methodologies, 1364 Competitive homogeneous assay format, CompudoseÕ implant
pharmacological evaluation, 1365 1567 controlled-release subdermal implant,
retro synthetic analysis, 1364 Complement dependent cytotoxicity 1086
solid-phase techniques, 1366 (CDC), 1137 cylinder-shape, 1086
solution-phase techniques, 1366 Complementarity determining regions diagrammatic illustration of, 1089
Combinatorial chemistry technology, 2486, (CDR), 1132 extrusion technique, 1086
2491 Complementary and alternative medical fabrication of, 1086
Comparative molecular field analysis practices, 67 polymer matrix diffusion-controlled, 1086
(CoMFA), 725 Complementary DNA (cDNA), 1420 Computational chemistry, 716
Commercial qualitative analysis software Complex-formation titrations, 3759 drug-receptor interactions, 716
introduction of, 3435 applications, 3760 electrostatic interactions, 717
Committee for Proprietary Medicinal endpoint detection, 3760 mathematical models, 717
Products (CPMP), 1594, 3999 equilibrium, 3759–3760 biological properties, 717
ad hoc groups, 1596 inorganic complexing reagents, 3760 physical properties, 717
decision, 1598 Complex carbohydrates, 2870 molecular mechanics, 720
of European Economic Community (EEC), Complex formation, 777 molecular simulations, 724
544 Complex mixtures, protein components of, Newtonian mechanics, 717
guideline, 1625, 2270 3042 non-deterministic theory, 717
pharmaceutical sponsors, 1596 Complexation and chemical modification, pharmacodynamic interactions, 717
working parties, 1596 for flavor modification technology, quantitative correlations, 716
biotechnology, 1596 1771 quantum mechanics, 717
efficacy, 1596 Compliance software package for, 3005
pharmacovigilance, 1596 shear, 3129 Computer aided drug discovery, 4013
safety, 1596 tensile, 3129 Computer-aided image analysis, 2583
Committee for Veterinary Medicinal Products Compliance and communication, 3981 Computer-aided molecular modeling
(CVMP), 1595, 1597 veterinary medicine, 3981 methods, 714
decision, 1598 Complimentary physicochemical techniques, Computer and communication aids, 739
working parties, 1596 advantage of, 1061 Computer applications, 732
efficacy, 1596 Compounding pharmacy monographs, classification of, 732
immunological veterinary products, 2852 Computer-assisted drug design (CADD), 714,
1596 Comprehensive Medicinal Chemistry (CMC), 725, 2808
pharmacovigilance, 1596 4015 in aircraft manufacturing industry, 715
safety, 1596 Compressed air–nitrogen pressure, 2881 Cambridge Crystallographic Data Center,
Common-ion effect, advantage of, 3004 Compressed pills. See Tablets. 726

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I28 Index

[Computer-assisted drug design (CADD)] Concentric cylinder viscometers, 3139 Connective tissue papillae, 2666
computer modeling, 725 Concomitant medication, 567, 1520 Conserves. See Confections.
equipment, 728 Concomitant polymorphs, 850 Constitutional isomers, 2142
macromolecular structures, 726 Condita geometric isomers, 2142
methodology, 727 confitures=sweet meats, 954 optical isomers, 2142
nuclear magnetic resonance (NMR) constituents of, 954 Consumer Health Products Association
spectroscopy, 725 Conditional tests, for animals, 1661 (CHPA), 2854
nuclear overhauser effect (NOE) signals, Conduction heating, mechanism of, 3514, Consumer Product Safety Commission
725 3869 (CPSC)
three-dimensional macromolecular Conductivity detectors, 533 drug packaging guidelines and regulations,
conformation, 725 Conductometric titrations, 3762 2526
pharmaceutical industry, 726 advantage, 3763 regulatory control for soaps, 800
pharmacophore model, 725 applications, 3763 Consumers Healthcare Products Association
advantages of, 726 acid–base titrations, 3763 (CHPA), 2414
hypothesis, 726 complex-formation titration, 3763 Contact charging
three-dimensional, 725 precipitation titrations, 3763 metal–metal, 1535
two-dimensional, 725 curves, 3763 for solid electrification, 1535
website reference for, 726 Cones, medicated, 954 Contact dermatitis, 72
software packages, 729 applications, 954 Contact lenses
theory, 727 Cone–plate viscometers, 3129, 3140–3141 classification of, 2202, 2203
guidelines for, 727 Confidentiality and Security controls, 2561 hydrophilic soft, 2202
X-ray crystallographic analysis, 725 Configurational isomers, 2142 rigid gas-permeable, 2202
Computer-based decision support system, Confitures. See Condita. drug release from, 1221
design of, 3992 Confocal microscopy techniques, use of, 1142 erodible polymeric delivery systems,
Computer-generated stability calendar, 735 Conformational isomerism 1222
Computer hardware technologies of cycloalkanes, 2148–2149 non-erodible systems, 1224
hardware=human machine interfaces, 709 in cycloalkyl and cycloheteroalkyl by Ocusert, 1222
network requirements, 709 structures, 2148 drugs delivery from, 1221
redundancy requirements, 709 of 3-methylfentanyl, 2149 from collagen shields, 1222
selection of, 709 Conformational polymorphism, definition of, enzymatic products for, 2205
Computer-processing equipment, 2936 hydrophilic, 1221
development of, 2587 Congealed carriers, properties of, 763–764 materials for, 2202
Computer program Congealed material plastic, 2202
examples of labels generated by, 733 micro- and macrostructure of, 770 polymeric materials, 2202
sample labels generated by, 734 stability and structure of, 769 for vision, 2202
sample stability protocol generated by, structural changes in, 771 Contact sensitivity, delayed, 122
735 structure analysis Contact times
Computer program output, evaluation of, by differential scanning calorimetry drug products, 3696
2766 (DSC), 770 functional definition, 3696
Computers by infrared spectroscopy, 770 mechanical definition, 3696
in pharmaceutical technology, 732–741 by X-ray diffraction (XRD), 770 Container–closure system
in pharmacokinetics, 138 Congealing applications, waxes in, 765 chemical additives, 1694
regulatory issues affecting use of, 741 Congealing device, design of, 763 chemical ingredients, 1694
Computers and data acquisition software Congealing processes, in pharmaceutical fabrication of, 1694
speed and capacity, 3688 industry, 761 marketing of, 1946
tablet press, 3688 Congestive heart failure, 216 plastic components, 1694
validation tests of, 3689 treatment of, GTN in, 1076 standards=acceptance criteria for,
Computer-simulated plasma concentration- Congresslonal Budget Office (CBO), 1895 1696
time curves, 1013 Conium maculatum, principles of, 255 USFDA policies for, 1696
Computer system validation and Code Conjugate acids and bases, ionic equilibria of, Container-closure integrity test, 2789
of Federal Regulations (CFR) 386 by bacterial challenge test, 383
part 11, 4 Conjugation principles of, 2789
Computer systems validation (CSV), 559 amino acids, 320 selection of, 2789
Computer technologies, application of, hepatic mitochondria, 320 sterile broth filled units, 383
2560 Conjugation reactions Continuing Medical Education (CME)
Computerized process-control systems, 740 acetylation, 319 programs, 60, 62
Computerized systems, inspection of, 2560 biotransformation, 317 Continuous ambulatory peritoneal dialysis
Computerized validation, 556 endogenous compounds, 317 (CAPD), 21
Computerized warehouses, implementation glucuronidation, 317 Continuous batch process cooker
of, 1944 glutathione, 318 advantages of, 2232
Concanavalin A, 1421 addition reactions, 9 components of, 2232
Concentration–effect relationship classification of, 318 disadvantages of, 2232
description of, 2804 substitution reactions, 318 installation, 2232
Concentration of solute in liquid phase sulfation, 318 preparation method, 2232

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I29

Continuous carrier irradiator, 749 Convection, 1436 Corn starch granules, scanning electron
Contraceptive steroids, synthetic, 942 forced, 3513 micrograph of, 3478
Contraceptive vaginal rings (CVR), natural, 3513 Corn syrup
1350–1351 in pharmaceutical industry, 3513 formation of amorphous glass, 2231
Contract manufacturing organizations heat transfer, 3869 pH of, 2231
(CMO), 2486 Convective fluid, dynamic viscosity of, physical properties of, 2231
Contract research organization (CRO), 1931, 1436 preparation of medicated candies, 2231
2486, 2876 Convective heat transfer Corneocyte protein envelope, 1311
Contract service organizations (CSO) equation for, 3513 Corneocytes, size of, 3821
clinical trial services of, 2496 process, 3513 Coronary artery disease, obstructive, 365
phases, 2496–2500 Convective heat transfer applications, Coronary blood flow, 365
consolidation and expansion, 2487 in pharmaceutical drying, 1436 Coronary perfusion, 347
development-assessment services potential Convective heat transfer coefficient, 1436 Coronary vasodilators, 1248
by, 2494 Convective mass transfer coefficients, 1439 Coronary vasospasm, 116
drug discovery services by, 2491 Convective mixing, mechanism, 2747 Corporate assets, protection of, 2556
for drug discovery, 2486 Convective thermal resistance, 1437 Corporate management, 14, 1383
history, 2486–2487 Conventional airflow, 2175 Corporate network security, 2556
identification strategies, 2488 Conventional recrystallization, Corporation’s document management
mega, 2487 of pharmaceuticals, 3572 systems, 2556
monitoring, 2489–2490 Conventional vaccines Corrective action plan (CAP), 2305, 2313
multiple and various clinical trial support inactivated, 3908 implementation, 2313
services by, 2498–2499 pathogenic organisms in, 3908 COrrelation SpectroscopY (COSY), 3446
regulatory affairs capabilities by, 2501 Cooking machines double quantum filtered, 3446
selection criteria, 2488 components, 2232 Corrosion
communication, 2489 kettle turning device, 2232 atmospherical, 793
flexibility and adaptability, 2489 standard method for, 2232 basics, 782
integrity, 2489 Cooling processes, in pharmaceutical control measures, 788
partnership, 2489 industry, 761 cleanliness, 782
project management skills, 2489 Coordination penicillin compounds rust-free surfaces, 782
quality assurance (QA) awareness, 2489 structure of, 700 stainless steel use, 782
research area and therapeutic experience, COPD. See Chronic obstructive pulmonary costs, 782
2489 disease. current and current density, 786
timeliness, 2489 Copolymer, definition of, 2925 definition, 785
training, 2489 Copolymerization and polmer modification, electrochemical nature of, 785
trends for, 2506 156 engineer’s role, 789
ContramidÕ technology, 999 Copolymers, lipophilicity of, 2930 extent, 786
Contrast variation, definition of, 1050 Copolymers of styrene and divinylbenzene, factors influencing
Contrast variation experiments, 1051 472 corrosion-resistant materials selection,
Contrheuma Gel Forte N, 1126 Copolymer surfactants, 1062 788
Control, manufacturing, and chemistry Copper, corrosion resistance of, 793 environmental influences on, 788
(CMC) processes, 2501 Coprecipitates and melts, 774–780 temperature effects on, 788
Control chart ball milling, 774 fatigue, 784
average chart, 3500 controlled precipitation, 774 Federal Highway Administration (FHWA)
C-Chart, 3503 definition of, 774 estimation of, 782
cumulative sum (or Cusum) control chart, fluid energy micronization, 774 forms of
3503 grinding, 774 cervice, 784
design, 3501 trituration, 774 erosion, 783
exponentially weighted moving average Coprecipitation, 2049 galvanic, 783
(EWMA) control chart, 3503 Copy DNA (cDNA), definition, 930 intergranular, 784
factors, 3500 Copyright protection, importance of, pitting, 783
individual—moving range chart, 3503 2604 selective leaching, 784
for multiple processes, 3502 Cordials, sweetened alcoholic preparations, uniform or general, 782–783
P-Chart, 3503 954 fretting, 783
Range chart (R-Chart), 3500 Core data management processes, 551 impact on chemical process industries
Run Charts, 3503 Core process, 3075 (CPI), 782
S chart, 3503 quality of, 3075 indications, 785
Shewhart chart, 3500 Cored dendrimer, synthesis, 883 mechanisms, 783
standard deviation chart, 3503 Corn oil, 1423 mitigation, 788
usage phases, 3503 Corn starch molybdenum in, 791
Control standard endotoxin (CSE), disintegrant properties, 3561 monitoring
control of, 3059 in pharmaceutical industry, 3476 by corrosion probes, 785
Controlled heat-assisted drug delivery unmodified by visual inspection, 785
(CHADD), 3851 moisture isotherm for, 3479 of metals in seawater, 783
Controlled release aspirin tablets, 2408 optical micrograph of, 3479 of pharmaceutical equipment, 782

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I30 Index

[Corrosion] [Cosmetics] [Cosolvents]

and polarization, 786–787 performance of, 803 rapid bolus injection of, 812
oxidation–reduction potentials in, 786 guidelines for documenting, 804 route of administration, 812
processes, 785 petrolatum in, 805 solubilization, 806
range in industries, 782 physical stability of, 804 estimation of solvent composition, 808
rate-controlling step within, 787 and cosmetic efficacy, 804 methods and theories, 806
reactions and cosmetic elegance, 804 as solubilizers, 811
oxidation of pure metal, 785–786 preservatives, use of, 804 stability of drug, 806
reduction, 785–786 warning statement on, 801 systemic effects of, 812
type and severity of, 786 water, role of, 804 topical products, 815
Corrosion-resistant materials Cosmetics and drugs, 798–805 types of excipient, 1623
aluminum, 787 Cosmetics, Toiletries, and Fragrance water-miscible organic solvents, 806
chromium, 787 Association (CTFA), 801 Cosolvents, pharmaceutical
ferrous metals, 789 Cosolvency cosolvent solubilization, 3321–3322
iron, 787 advantages of, 2220 n-octanol, 3319
molybdenum, 790 chemical potential, 2220 physical properties of, 3319
nickel, 787 cosolvents, 2220 polarity and physical properties of, 3320
performance, 782 advantages of, 2220 solubilization profile
selection definition, 2220 for complexation, 3327
stainless steel, 789 free energy of molecule, 2220 surfactants, 3326
steel, 789 oral liquid formulations, 2220 in water, 3318
titanium, 787 solubility of volatile constituents, 2220 Cotransporters, 26
Corrosion resistance, improvement of, 4046 water-miscible organic solvents, 2220 mechanism of, 27
Corticosteroid, 171 disadvantages of, 806 sodium–glucose, 27
intramammary infusion in lactating cows, drug solubility, 2220 Coulometric detector, 535
3959 limitations of, 2220 Coulometric titrations, 3764
Corticosteroid betamethasone, 2633 liquid drug preparations, 806 advantage, 3764
Corticosteroid budesonide, 2980 solvent polarity, 807 application, 3764
Corticosteroids, 983, 2483 therapeutic agents complex formation titrations, 3764
effect of, 1256 lipophilicity, 2220 neutralization titratrions, 3764
Corticosteroids triamcinolone acetonide, structural complexity, 2220 oxidation–reduction reactions, 3765
1319 Cosolvents, 2861 precipitation titrations, 3764
Cortisol, in plasma and urine, 545 advantages of, 806 endpoint detection, 3764
Cortisone acetate in small-volume parenteral preparations, primary technique, 3764
delivery of, 185 817 secondary technique, 3764
solid drug particles of, 1220 biologic effects of, 806, 812 sources of error in, 3764
Cortisporin, 983 definition of, 806 Coulter counter, operation of, 2589
Corundum (a-Al2O3) degradation of drug, 806, 1623 Coumarins, 2911
inorganic compounds, 4110 dilution of formulations, 811 Council on Codes and Standards (CSS),
Cosmetic Act, 1779 in drug formulations, 806 2238
Cosmetic elegance, 804 fraction, 811 Counter-ion adsorption, in Stern layer,
Cosmetic, excipients for, 1638 hemolytic effects of, 816 4121
Cosmetic industry–physician–user hemolytic potential of, 815, 1623 Countercurrent methods, 4
relationship, 805 dynamic flow conditions, 815 disadvantages, 4
Cosmetic Ingredient Review (CIR), 801 static flow conditions, 815 phase–volume ratio, 4
Cosmetic ingredients, 799, 802 impurities, 817 Counterfeit drugs, detection of, 4100
description of, 803 interpolation method for, 809 prevention of, 4100
and photosensitization, 803 intramuscular (IM) injection, 812 Counterfeit excipients, use of, 2779
restriction of, 801 intravenous (IV) infusion fluids, 811 Counterion
safety of, 801, 803 irritation data, 812 hydrogen-bonding nature of, 3179
shorthand nomenclature for, 803 linear function of, 809 hydrophilicity of, 3184
Cosmetic manufacture, guidelines for, 804 liquid dosage forms Counterion transport, direction of, 2747
Cosmetic preparation, 2980 ophthalmic and otic dosage forms, 817 Cowpea mosaic virus, 3912
Cosmetic regulations, violatorsof, 799 oral dosage forms, 817 CpG DNA, 3922
Cosmetic suspensions, 3605 parenteral dosage forms, 817 Cranial cavity, 2681
Cosmetics topical dosage forms, 817 Controlled-release drug delivery systems
adulteration of, 804 in mice and rats, 812 (CrDDSs), 1082
chemical stability of, 803, 804 myotoxicity of, 816 Creams
as emetics, 804 ophthalmic products, 815 advantages, 996
label information, 803 osmotic effects of, 817 classification of, 1082
microbiologic requirements, recommended, otic products, 815 cold, 996
804 parenteral products constituents, 813 drug formulation, 1082
microbiological contamination of, 804 polarity, 808 drug molecules, 1083
penetration into skin, 805 and polarity indices, 806, 809 medicated solid, 954

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I31

[Creams] Crospovidone MÕ, 3556 [Crystal(s)]

microstructure of, 1126 Crossflow stream, sweeping flow of, 4041 nucleation, mechanisms for, 838
of o=w type, 1127 Cross-Ject, 1217 pharmaceutical, design of, 615
of w=o type, 1127 Cross-linked protein crystals phase transitions of organic, 852
rate constant, 1083 antigens, 3922 potassium ferricyanide, 829
definition of, 1083 Cross-polarization (CP) dynamics prismatic, 847
rate-preprogrammed, 1083 active pharmaceutical ingredient, 3304 rod-shaped, 829
semisolid emulsions, 954 Cross-polarization magic angle spinning structure and nomenclature, 845–846
transdermal, 1086 (CP–MAS), 3456 sulfamethoxazole, 830
transmission electron micrograph of, 1127 spectra of lamivudine, 3457 surface, roughness of, 844
vanishing, 996 Crossover design, for drug products, 172 trimethoprim, 830
Creatine phosphokinase, 364 Crotid atherosclerotic lesions, 1160 valence, 3545
measurement of, 118 in vitro in cell cultures, 1160 Crystal Data cell, 4105
Creatinine clearance, 2636, 2819 Croton oil topical inflammation test, 116 Crystal faces, types of, 848
endogenous, 1021 Crown ethers, 2160 Crystal forms, existence of, 2935
Creatinine concentration, 574 chiral recognition properties of, Crystal growth, by heat transfer, 3885
Credible new drug applications (NDAs), 2160–2161 Crystal lattice, hydrate, 1823
2877 chromatographic procedure for, 2161 Crystal morphology, change of, 41
Creep curve, mechanical model of, 3136 Crude drug, making of, 2905–2906 Crystal nucleation, 40
Creep test, 3135 Crude Drugs Association of Tokyo, 2837 Crystal polymorphic transformation,
Cremophor EL, 3349 Crushing strength, 3633 kinetic coefficient of, 42
emulsifying agents, 1624 definition, 3633 Crystal polymorphism, 1823
Cremophor EL and ethanol, combination of, Erweka hardness tester, 3633 Crystal shape, feedback control of,
3361 testing of mechanical strength, 3633 861
Cremophor RH40, 3349 types of hardness testers, 3633 measure of, 869
CremophorÕ, 1258, 3336 Cryogenic milling Crystal structure, nature of, 2935
Crenation, 1272 for heat-sensitive compounds, 2344 Crystal structure disorder, on solubility and
Crepe, 1466 nonflammable refrigerant, 2344 dissolution rate, 3313
Crest tartar control dentifrice, 899 Cryopelletization Crystal surface area, 868
Crest tartar control mouthrinse, 899 cross-linked biopolymers, 2661 Crystal violet lactone, 2316
Creutzfeld–Jakob Disease (CJD), 1596, 3997 droplet liquid formulation, 2661 Crystalline
Crevice corrosion, 784 drug-loaded pellets, 2661 pharmaceutical materials, 89
CRF. See Case report forms. equipment components, 2661 polymorphs, 90
Cricopharyngeal reflex, 2867 formulations-related variables, 2661 Crystalline a-lactosemonohydrate
Crinone gel, 984 immediate-release formulations, 2661 as tablet diluent, 3681
Critical association concentrations (CACs), solutions=suspensions, 2661 Crystalline anhydrous glucose, near-infrared
2921 spherical droplet formulations, 2661 spectra of, 3378
Critical binding reagents Cryoprotectants, 1613, 1826 Crystalline bulking agents, polymorphism in,
development of monoclonal antibody, Cryoprotection 1823
1575 composition of, 1630 Crystalline compounds, purity of,
polyclonal antibodies for, 1575 mechanism of, 1630 397
Critical flocculation concentration (CFC), trehalose used as, 1630 Crystalline drug, solubility of, 41
4123 Crypts, definition of, 2714 Crystalline drugs
Critical fluid liposomes (CFLs), 3579 Crystal(s) amorphous content of, 398
Critical micellar concentration (CMC), 35, appearance of, 847 polymorphic forms of, 89
292, 1824, 2222 benzamide, intermolecular interactions in, Crystalline excipients, 1648
Critical relative humidity 617 polymorphic forms of, 89
for water-soluble components, 4057 classification of, 3545 Crystalline lactose, 35
mixture solution, 4055–4056 effectiveness of, 841 Crystalline mannitol, 1809, 1810
Crohn’s disease, 218, 1228 growth, 843–844 Crystalline materials
patients with, 2872 experimental strategies, 844 and amorphous materials, 2079
Cromolyn sodium, moisture uptake, 4057 mechanism of, 844 characterization by X-ray diffraction
Croscarmellose, capsule formulations, 415 rates of, 844 (XRD), 2078
Croscarmellose sodium (Ac-Di-SolÕ), 1248, habit, 628–831 milling of, 2078
1291, 2454, 3553–3554 factors influencing, 821 Crystalline powder, 3885
flow properties, 3554 human insulin, surfaces of, 845 Crystalline salt, pharmaceutical, 616
scanning electron photomicrographs of, ionic, 3545 Crystalline solids, 1115, 1808, 4103
3555 lattice defects, 826, 3544 phases of fluprednisolone, 4104
Crospovidone, 3554–3555 metallic, 3545 water sorption by, 4054
coarser grades of, 3560 molecular, 3545 X-rays on, 4103
particle sizes of, 3556 morphology, 844–845, 846 Crystalline states
scanning electron photomicrographs of, prediction and other computational liquid, 1115
3556 strategies, 847–849 long-range order in, 1115–1116
water wicking in, 3555 in pharmaceutical processes, 846–847 short-range order in, 1115–1116

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I32 Index

Crystalline structure Cyclazocine, and biodegradable polymers [Cyclodextrin]

collapse of, 1441 drug delivery, 177 in skin permeation, 1313
on solubility, 3311 Cyclic amines, titration, 3756 solute inclusion complexation, 2158
Crystalline sucrose, limits of detection and Cyclic carboxamide dimers, 617 structure, 2158
quantitation, 4104, 4108 Cyclic glucose polymers, 1617 use of, 15
Crystallinity, 3434, Cyclic polyethers, 1507 Cyclodextrins (CDs), use of, 2776
of pharmaceuticals, 4108 Cyclic voltammetry, technique of, 1494 Cycloglucans, 682
Crystallization, 398, 834–835, 853 Cyclization–elimination, reaction of, 3009 Cyclohexane, radiolysis of, 3544
of buffer components, 840 Cyclization–elimination sequence, 2997 Cyclohexanes conformation isomerisms
definition of, 858 Cyclo-olefin copolymer (COC) boat, 2148–2149
degree of, 1810 capsule, 1213 chair, 2148–2149
energy of, 849 in drug storage, 1213 twist boat, 2148–2149
kinetics, role of, 834 Cyclo-oxygenase-2 (COX-2), 2435 Cyclohexylamine formation, 1616
of paracetamol polymorphs, 835 Cycloaddition reaction, 2997 Cyclooxygenase-derived prostaglandins,
pathways, implications for, 843 Cycloamyloses, 682 3054
of polymorphs, 835 Cyclobutyl dimmer, formation of, 620 Cyclophosphamide, 363
solvent effects on, 849–851 Cyclodextrin optical isomers of, 2144
principles of, 862 absorption, 685 Cyclosporin, 1610
inhibition, 41 advantage of, 2701 benefits of, 426
inhibitors, 43 characteristics of, 682 delivery of, 2750
kinetics, 865 complexation, 671 Cyclosporin (CSA)
determination of, 866 benefits of, 680–682 administratrion of, 2743
devolopment of, 866 improvement in solubility, dissolution, Cyclosporin A, 3348
Crystallization processes, 88, 834, 870 and bioavailabilty, 680–681 with nanospheres, 1192
complexity of, 821 improvements in drug stability, 681 oral bioavailability of, 3349
model for, 862 reduction in volatility, 681–682 plasma concentrations of, 349
monitoring of, 860 reduction of unpleasant side effects and wound modulator, 1223
solvent bitter taste, 681 Cyclosporin nanocapsules, preparation of,
viscosity of, 820 constituents of, 2158 1196
Crystallization model, simplification of, derivatives, 683–685, 686 Cyclosporine
866 hemolytic effects of, 688 bioavailability of, 1257
Crystallization models, derivation of, 870 hydroxypropyl, 684 immunosuppressive, 2737
Crystallization of bulking agents, methylated, 683–684 in organ transplantation, 2737
conditions for, 1823 sulfobutyl ether, 684–685 preparation of controlled release, 780
Crystallization of excipients and distribution, 685 Cyclosporine (immunosuppressant),
consequences, 1809–1810 effects of charge 2254
Crystallization solvent, metastable form of, density, 678 Cyclotrons, 3090
2939 proximity, 678 Cylindrical epithelial cells, layer of,
Crystallization technique, parallel, 3004 state of, 678 2717
Crystallizing solvent, nature of, 824 excretion, 685 CYP-450 3A isoenzyme, 2909
Crystallographic classification, 821 factors affecting, 675–680 CYP2C19
Crystallographic twinning, 826 steric effects, 675–677 Japanese population, 322
Cucufa, 954 functional groups of, 1618 CYP2C9 polymorphisms, 2198
Cullet, 2509 inclusion complexes with, 3579 CYP2D6
Cup and bob viscometer, principle of, inclusion, 671–674 Caucasian population, 322
3139 evaluation of, 674–675 consequences of, 1898
Current good manufacturing practice ionizable, 1258 drug-metabolizing P450 cytochrome,
(CGMP), 591 mechanism of solubilization by, 1617 1898
applicability of, 1942 metabolism, 685 mutations, 1898
regulations of, 1941 as mobile-phase additives, 2158 nicotine metabolizing cytochrome,
regulations of, 2783 natural 1899
sections of, 2783 drug conjugates of, 1255 CYP3A4 induction, 2194
training in, 1943 in oral solutions, 3350 CYP (Cytochromes P450)
Cutaneous applications, powders for, 2979 parent, 1617 drug biotransformation
Cutaneous blood flow, 3822 pharmacokinetics in enzyme systems, 321
blood vessels oral, 685 Cysteine, oxidation of, 285, 1825
accessibility of, 3822 parenteral, 685 Cysteine residues, labeling of, 3045
cutaneous perfusion, 3822 properties of, 1618 Cysteinyl-leukotriene inhibitors.
demonstration of, 3822 regulatory status, 690–691 See ACE-inhibitors.
systemic absorption, 3822 retention properties, 2158 Cystic fibrosis, 2093, 3037
Cutaneous gene delivery, 2752 safety of, 685–690 patients with, 2872
Cutina HRÕ, 765 oral, 685–687 treatment of, 2704
CutinaÕ, 1258 parenteral, 687–689 Cystic fibrosis (CF) IndicatorÕ, 2119
Cyclamate, 1616 reproductive, 689–690 Cytarabine release, 2033

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I33

Cytochrome c reduction assay, 151 DaunoXome, 1128 Dementia, Alzheimer’s-type, 73

Cytochrome enzyme system Deaeration theory, 3169 DemerolÕ, 1040
(CYP450 system), 1911 Deamidation, 1826 Demixing
Cytochrome P450, 71, 1610 of asparagines, 1827 types
catalysis, mechanisms of, 312 reactions, 283 axial, 2358
chromatographic and immunologic Deborah number (De), 2031 competitive patterned, 2358
behavior, 312 Debrisoquine radial, 2358
in drug biotransformation, 312 hydroxylation of, 1018 D-enantiomeric forms, in biotechnology drug
in endoplasmic reticulum, 312 metabolism, 1898 substances, 461
in hepatic enzymes, 1398 Decamethylcyclopentosiloxane (DMCPS), Denaturation
lung, in Clara cells, 322 606 cold, 1810
for metabolism, 1398 DecapeptylÕ, 177, 178, 607 protein
multigenic family of heme proteins, 312 for prostate cancer treatment, 178 cryoprotectants, 1275
substrate specificity, 312 and serum LH and testosterone freeze-drying, 1275
as terminal oxidase, 312 concentrations, 178 lyoprotectants, 1275
Cytokine immunoassays, caliberation of, Decipher receptor gene function, 3125 types of substances, 1275
4101 Decoctions, 954 DendrilockÕ structure, 881
Cytokine therapy Decoctum. See Decoctions. Dendrimers, 1063
in inflammatory bowel disease, 3923 Decubitus ulcers, 1028 architectural components, 872
in multiple sclerosis, 3923 Defecation, suppression, 2871 applications of, 877
in rheumatoid arthritis, 3923 Deflagrations and explosions, 2882 biological, evaluation of, 886
Cytokines Deflocculants and dispersing agents, biological properties
coadministration of, 3914 of suspensions, 3606 biodistribution, 886
in keratinocyte differentiation process, Deformation, 3128 immunogenicity, 886
1311 plastic, 1736 toxicity, 886
Cytolytic mechanism, 1139 Defuzzification, 2404 branch cell components, 875
Cytomegalovirus (CMV), 1224 Degradation cancer therapy on, 884
Cytopasmic peptidases, 2718 aliphatic polyphosphate vs. aromatic carbosilane-poly(ethylene oxide), 886
Cytoplasmic machinery, plasma cell with, polyphosphonate, 190 charge interactions and aggregation, 876
1133 and absorption, 19 delivery DNA, 884
Cytoplasmic membrane, 2992, 3550 biodegradable polymers, 176 delivery systems of, 884
Cytoplasmic peptidases, 2722 chemical, 1827 differential scanning calorimetric (DSC)
Cytoskeleton-specific immunoliposomes DNA, 3912 measurements, 878
preservation of cell viability by, 1160 drug, 165, 1623 drug carrier, 884
with hypoxic cells, 1164 and drug biotransformation, 310 flow properties of, 878
Cytosolic leucine aminopeptidase, 2674 in drug delivery, 177 fractured, 884
Cytostatic drug biotransforms, 699 excipient, 1641 functional terminal groups, 876
Cytotoxic affect, 1133 monomers in, 176 growth of, 872
Cytotoxic drug compounds, 419 oligomers in, 176 hyperbranched polymers, 886
Cytotoxic drugs, 1327 protein, 1274 hydroxyl surface
Cytotoxic effects, of fatty acids, 17 random chain scission, 176 in dilute solutions, 877
Cytotoxic radiation, 1138 rate of, 389 hydrodynamic properties, 877
Cytotoxic T lymphocyte antigen-4 (CTLA-4), stereospecific biotransformation, internal structure, 877
3922 321 macromolecules, 872
Cytotoxicity issues, 687, 688 hydrolysis of polymer chains, 176 molecular modeling, 874
Dehumidification techniques, 1444 oligonucleotides, 884
Dehydroemetine, 1191 peptide, 874
Dahlinder’s repose-angle device, 3282 Dehydroepiandrosterone, 71, 2431 permeability of, 883
Daily cleaners effects, 2439 pharmaceutical applications of, 879,
oxytetracycline in, 3956 for mental health, 2440 886
with shearing particles, 2204 Dehydropregnenolone acetate, 245 physicochemical properties, 874
surfactants, 2204 Deionization system, continuous, 4041 poly(aryl ether), 874
D-alpha-tocopherol, 3348 Delivered-dose uniformity poly(benzyl ether)), 874
DanatrolÕ dissolution profiles, 2820 determination, 2087 poly(phenylacetylene), 874
Danazol delivery, 1349 FDA expectations for, 2087 poly(propyleneimine), 874, 877
Danger signals, dendritic cells, 3914 Delivering medication, method of, 3979 polydispersity, 884
Darodipine–polyethylene glycol, phase DELSA 440 instrument, 4119 polyelectrolyte, 876
diagram, 3744, 3746 electrophoretic mobility distribution, polystyrene particles, 883
Darvocet, 2426 4119 size and shape, 875
Daunomycin conjugates, construction of, laser-scattering particle electrophoretic solubilizing power, 879
1136 analyzer, 4119 solutions
Daunomycin targeting, conjugates of, 1331 zeta-potential distribution, 4119 bulk flow properties, 879
Daunorubicin, 1332 Deltoid injection, 2646 viscosities of, 879
Daunorubine, cytostatic, 1128 Deltoid tuberosity, 2646 structural components, 872

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I34 Index

[Dendrimers] Deramciclane, absorption of, 2821 [Detector]

synthesis Derivative spectroscopy, advantages of, thermal, 3409
advantages of, 872 3472–3473 UV absorption, 1522
convergent method of, 872 Dermabrasion, 1028 UV–vis spectophotometric, 534
transfection efficiency of, 884 irritation scoring system, 122 variable wavelength, 534
zero-generation (G0), 872 toxicity studies, 2774 Deterioration fixed oils, 1624
Dendrimer–drug complexes, 881 and transdermal drug delivery, 1311 Dethylenetriaminepentaacetic acid ligand,
Dendrimer surface density, 881 Dermatological drug delivery system, 1313 1138
DendriporeÕ structure, 881 Dermatological formulations, 1321–1322 DETOU polyorthoester
Dendritic cells, 1281, 3914 bioequivalence of, 1321 degradation of, 186
Dendritic structures in vivo tape stripping technique, 1321 naltrexone pamoate release from, 186
atomic force microscopy (AFM), 875 advantages, 1322 reaction of 1,2,6-hexanetriol with, 186
cavity size of, 879 pharmacokinetic parameters, 1322 Detoxification mechanisms, 120
entrapment effect, 880 Dermatological gels, 1887 Deuterated material, hydrogeneous material
gastrointestinal tract, 883 Dermatological preparations, antibacterial for, 1051
hybrid, 879 agents, 3971 Deuterated polymer chains, 1064
hydrophilic surface layer, 879 Dermatological therapy, 1317 Deuterated propylene glycol, 1061
hydrophobic core, 879 Dermatologic laboratories in cosmetic Deuterated triglycine sulfate (DTGS), 3376
Dendritic unimolecular micelles, 879 safety, 805 Deuterium oxide dispersion medium, 1066
solubilization capacity of, 880 Dermatologic products, packaging of, 3270 DeVilbiss 646, 2097
water-soluble, 880 Dermatologic vasoconstriction, drug DeVilbiss aerosonicÕ, 3859
Denileukin, 272 products, 171 DeVilbiss pulmosonicÕ, 3855
Densification Deryaguin–Landau–Verwey–Overbeek Ultra-Neb 99Õ ultrasonic nebulizer, 3859
degree of, 829 (DLVO) theory, 4121 ultrasonic and jet devices, 3857
segregation, 2975 electrostatic stabilization, 4122 Devitrification, 2509, 2510
Density functional theory stability of oil-in-water emulsions, 1557 Dewar flasks, 3398
chemical energy, 719 stability of water-in-oil emulsions, 1557 Dexamethasone, 2119
chemical structures, 719 Desacetylcephalosporin C biosynthesis of, Dexamethasone poly (L-aspartate), 1233
Kohn–Sham equations, 719 255 DexonÕ, 177, 1223
Möller–Plesset (MP2) theory, 719 Des-enkephalin-gamma-endorphin, 2728 Dextranase, 2035
Dental liniments, 960 Desensitizing agents, 896 Dextran, 1630, 1184
Dental plaque, 2442 Desert tea, 74 hydrogels, 1237
Dental products, 891 Desglycinamide arginine vasopressin, 2728 drug delivery system, 1644
Dentifrices, 893, 954 Desktop publishing package, 554 Dextranomer, in polyethylene glycol 600
Dentifricium. See Dentifrices. Deslorelin–PLGA microparticles, scanning (PEG 600), 1031
Dentilimentum. See Dental liniments. electron microscopy (SEM) of, 3578 Dextrins, flow characteristics of, 3286
Deodorizing dressings, in wounds, 1028 Desmosomes, 1340 Dextromethorphan, 2416
Deoxycholate, 16 Desolvation effects, 717 Dextrose
Deoxycorticosterone acetate (DOCA), 116 Destabilization, 2861 advantage of, 1613
Deoxyhemoglobin, 357 Detector tonicity adjusters, 1627
solutions, 356 amperometric, 535 Dextrose equivalent (D.E.) value, 3481
Deoxyribose nuclease, human, 1428 applications, 3467 Density functional theory (DFT), 719
Depakote SprinkleÕ, 1771 bulk property, 533 D-glucono-d-lactone, 2027
Department of Health and Human Services cell designs, 1500 D-glucose monomers, composition,
(DHHS), 1779 characteristics, 3464 cryoprotection, 1630
DepoCytÕ, drug product, 1643 charge coupled device, 3464 Diabetes
DepoFoamÕ, drug product, 1643 conductivity, 533 management of, 1616
Depolymerizable polymers, 159 coulometric, 535 type-II, 2705
soluble, 159 diode array, 534 Diabetes mellitus, 3037
water-insoluble, types of, 159–160 dual-electrode, 1522 protein for, 267
Depolymerizable water-insoluble polymers, electrochemical, 534–535 Diabetic gastroparesis, 2872
160, 161 evaporative light scattering, 535 Diabetic neuropathies, 72
DeponitÕ system fixed wavelength, 534 Diabetic ulcers, 271
cross-sectional view of, 1090 fluorescence, 534 Diachylon plaster, 1023
nitroglycerin releasing, 1087 high-performance liquid chromatographic Diacyl phospholipids, 976
structural components, 1090 (HPLC), 533–535 Diagnostic radiopharmaceuticals, 1267
Depot (or Projectile) needle-free injection in spectrophotometer, 3464 2,4-Diaminotoluene, 1514
by Caretek medical, 1210, 1217 photodiode array, 3466 Diaphragm valves, 4047
disadvantages, 1210 properties, 3408 Diarrhea, treatment of, 178
drug quantum, 3408 Diaspirin cross-linked hemoglobin, 355
delivery device, 1210 refractive index, 533 Diastereoisomeric chelates, 2159
formulation, 1210 sensitivity, 3408 Diastereoisomeric ion pairs, 2159
Depyrogenation, 3060 solid state, 3464 Diastereomeric derivatization, 2156–2157
dry heat, 3512 solute property, 533 Diastereomers, 2145

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I35

Diazepam (ValiumÕ), 1042 Dies [Differential scanning calorimetry]

anxiolytic properties of, 311 instrumented, 3787 sublimation, 3727
injection, by intravenous injection, 3955 manufacturing tolerance, 3787 for frozen systems, 4113
serum concentrations, 2632, 2645 nickel–chromium–boron alloying on, 3792 for lipid analysis, 981
Dibasic calcium phosphate, 988, 2772 tapered, 3785, 3787 heat flow measurement, 394, 3726
diluent in tablets, 3655 Die swelling, 2006 heat flow ways, 3727
Dibasic compound, solubilization, 3316 Diet and Reinfarction Trial (DART), 2439 heat flux, 394–395
Dibucaine hydrochloride solution, 1630 Dietary antioxidants, 142 high speed, 396
Dicalcium phosphate dihydrate, 43 Dietary fluoride, 892 measuring methods, 3726
agglomerates, 3229 Dietary proteins, absorption of, 296 heat flux DSC, 3726
differential scanning calorimetry of, Dietary supplement, 2850 power compensation DSC, 3726
2376 consumption of, 2908 melting temperature, 397
exposure, 2376 definition of, 2904 microwaves, 3729
in gastric juice, 3680 standardization of, 2906 modulated temperature, 394–395
hydroxyapatites, 3230 Dietary Supplement Health and Education for pharmaceutical applications, 3726
Karl Fisher titration for, 2377 Act (DSHEA), 68–69, 1789, 2841, for pharmaceutical materials, 396
milled form, 2377 2903 in pharmaceutical sciences, 393
unmilled form, 2377 Diethylene glycol, in kidney and liver damage, power compensated, 394–395
thermal gravimetric analysis of, 2374 1406 pressure, 3727
Dicalcium phosphate-based tablets Di-(ethylhexyl)phthalate, flux enhancer, 1222 principle, 3726
disintegrating pressure versus disintegration Diethylenetriamine pentaacetic acid, 3095, purity analysis, 3739
time of, 3360 858 advantage, 3740
disintegration of, 3358 Diethyl ethylene malonate, 1307 van’t Hoff’s law, 3739
water uptake versus disintegration time of, Diethylphthalate (DEP), 1802 reversible transition, 3735–3736
3360 Difference spectrophotometry, 3473 scan
Dicalcium phosphate matrix tablets, 3561 for determination of drug substance, 398
Dicaphos AN, 3229 biological matrices, 3473 glass transition temperature, 400
anhydrous type, 3229 constituents in tablets, 3473 of sucrose, 397, 400
hydroxyapatites, 3230 pharmaceutical preparations, 3473 trihydrate, 3738
Dichloromethane and hexane, advantage of, plant extracts, 3473 second-order transitions, glass transitions,
468 syrups, 3473 3727
Dichoism, circular, method of, 703 in turbid or cloudy solutions, 3473 signal
Dichotomy, biological activity of, 3054 Different X-ray powder diffraction, 2943 components, 3728
Diclofenac, 1125 Differential pulse polarography=voltammetry non-reversible events, 3728
formulation of, 2810 (DPP=DPV), 1495, 1496 reversible events, 3728
release mechanism, 2018 Differential pulse techniques, 1497 Differential scattering cross-section, 1053
salts Differential scanning calorimetry, 1809, 2938, Differential thermal analysis, 395, 701, 3726
diclofenac deanol (DDNL), 3181 2941 for pharmaceutical applications, 3726
diclofenac tert-butylamine (DtBA), advantages, 3727 Diffracted light signals, 2587
3181 amorphous content determination by, Diffraction grating, 3397
intrinsic dissolution rates for, 3178 3737 Diffraction patterns, generation of, 2587
melting points for, 3178 applications of, 397 Diffuse reflectance accessories
solid dosage forms of, 3183 calibration, 3728 types, 3377
solubilities for, 3178 combination with, 1052 for spectrum recording, 3381
solubility versus pH, 3179 conventional, 395 Diffuse reflectance near-infrared spectra
Diclofenac diethylamine, 1125 liquid excipient, 3746 for monosaccharides, 3379
Diclofenac sodium, 2010 phase diagrams, 3743 of pharmaceutical compounds, 3379
dissolution behavior of, 768 phthalylsulfathiazole, 3734 Diffuse-reflectance spectroscopy, 3414
granules of, 767 polyethylene, 3742 accessories for, 3375–3376
release, 2408 definitions of, 397 applications of, 3383
Diclofenac sodium=carnauba wax matrices, of dicalcium phosphate, 2376 bifurcated fiber bundles in, 3375
2010 drawbacks, 4113 caffeine by, mid-infrared spectra of, 3380
Dicumarol for drug-excipient compatibility studies, crystalline powder, 3380
dispersion, freeze–drying process, 775 401 crystalline urea, 3379
solid dispersion of, 779 drug purity by, advantages of, 398 instruments for, 3375
solubility of, 2870 dynamic, 394–395 KBr powder, 3382
Dicyclohexyl carbodiimide, 1135 endothermic transition, 3734–3735 KCl diluted samples, 3380
Didanosine, human bioavailability of, enthalpy of fusion, 397 Kubelka–Munk (K–M) function, 3377
2821 eutectic-point detection, 3740 limitations of, 3380
Didodecyl dimethylammonium exothermic transition, 3734–3735 linearization function, 3381
bromide(DDAB), 1065, 1564 first-order transitions matrix materials for, 3381
Dielectric constant, of pharmaceutical boiling, 3727 optical absorption characteristics, 3378
materials, 1448 crystallization, 3727 optical filters in, 3375
DiemulsÕ, 1550 melting, 3727 in pharmaceutical applications, 3383

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I36 Index

[Diffuse-reflectance spectroscopy] DilaudidÕ, 1040 Diphosphopyridine nucleotide-diaphorase

powders and bulk materials, 3381 Dilauroyl-sn-glycerophosphocholine (DLPC), (DPND), 1341
radiation sources in, 3375 1065 Diphtheria toxin, 1139
scattering characteristics, 3378 Diltiazem hydrochloride, 2010, 2012, 2034 Diphtheria toxoid, 2746
for trace analysis, 3382 release, 2013 Dipolar coupling, 3298
undiluted substances, 3379 Diltiazem immediate release tablet DiprivanÕ, 1270, 1550
with grating monochromator, 3375 formulations, 2407 dose of, 3360, 3362
Diffusely reflected radiation, for analytical Dilute aqueous systems, kinetic studies of, Diptheria–tetanus–pertussis (DTP) vaccines,
applications, 3377 3543 3915
Diffusion Diluted acids. See Acids, diluted. Direct chemiluminescence immunoassays
characteristics of, 26 Dilution determine test sensitivity, 3060 chemiluminescent molecules, 2058
electrode surface by, 1491 D-mannitol chemiluminescent properties, 2058
Fick’s laws, 1082 crystallization behavior of, 3307 immunological properties, 2058
mechanism, 26 in lyophilized formulations, 3306 Direct chiroptical detection, 453–454
membrane permeability, 26 in pharmaceutical formulations, 3306 Direct compression method, 3662–3663,
molecular, 343 polymorphic forms of, 3306 3673
transport system, 26 Dimercaprol (BAL), 702 active ingredients, 3674
Diffusion cell Dimethyl acetamide, 1623 ascorbic acid, 3663
experiments, phase in, 2743 Dimethylallyl pyrophosphate (DMAPP), paracetamol, 3663
models, 1303 239 pregranulated form, 3674
procedure for, 1302 Dimethyl beta-cyclodextrins, 16 advantages, 3674
Diffusion coefficients, 1172 Dimethylether (DME), 2270 anhydrous dibasic calcium phosphate, 3680
increase of, 1299 properties, 2271 calcium triphosphate for, 3680
Diffusion-controlled systems, control delivery Dimethylformamide, 2943 diluent, 3662–3664
for, 1248 Dimethyl sulfoxide (DMSO), 2437, 2743, disadvantages, 3674
Diffusion current, limiting, concept of, 1491 3350 disintegrating agent, 3662
Diffusion testing as concominants, 2447 glidant, 3662
bubble point pressure, 1755 Dimeticone emulsion hydroxyapatite used for, 3680
Fick’s law, 1755 frothy bloat in cattle, treatment of, 3950 limitation on use of, 3674
multipoint, 1754 oral liquids, 3950 process of, 3662
diffusive flow graph, 1754 Dinitrochlorobenzene, 122 tablet diluents in, 3675, 3678
filter’s performance, 1754 Dinitrophenol, 1406 for tablet manufacture, 3673, 3662
Diffusional boundary layer, 823 cataracts by, 1407 tricalcium orthophosphate for, 3680
Diffusional pathlength, 2666 uncoupling oxidative phosphorylation, Direct potentiometric measurements, 1505,
Diffusional resistance, 2666 1407 1509
Diffusive mixing, for pharmaceutical DinoprostoneÕ vaginal insert, 999 Direct surface sampling, leaned equipment
materials, 3898 Dinucleotide structures surface, 1588
Diflucan (antifungal), 2255 morpholino analog, 931 Discotic mesophases
Digested dietary proteins, 2717 normal DNA or RNA, 931 cholesteric, 1115–1116
Digestive absorptive unit, 2715 peptide nucleic acid, 933 columnar hexagonal, 1115–1116
Digestive system, 2713 phosphorothioate analog of DNA, 931 nematic, 1115–1116
Digital identification technologies, 2562 Dioctylpthalate (DOP), 2174 smectic, 1115–1116
Digital nervous system, 2558 Diode array detection (DAD), 94 Discrete particle counter (DPC), 2184,
Digoxin, 1922, 2045, 3349 Diode array detectors, 99, 534 2311
absorption, 2636 Diode array=rapid scan spectrophotometers, Disease management, pharmaceutical
chemical stability of, 778 3471 approach to, 1309
constituents of, 2255 Diode array spectrometers, 3375, 3438 Disinfecting contact lenses
intravenous injection solution formulation Diode array spectrophotometers, 3464 bacterial particle size of, 2206
for, 1670 Diode array spectrophotometric detector, fungus particle size of, 2206
pediatric elixir, 2255 1500 ocular irritation, 2206
quinidine-induced, 1399 Dioleoylphosphatidyl ethanolamine, 1065, product for, 2206
in renal tubules, 1399 1141, 3149 red eye, 2206
tablets BP, 917 endosomal compartment, 3149 thermal versus chemical, 2206
tissue binding of, 3030 lipid component, 3149 Disintegrant, super
toxicity, 2255 membrane destabilization, 3149 classification of, 3554
Dihydrofusidate derivatives, 15 DiPacÕ, 988 croscarmellose sodium, 3553
Diisopropylethylamine (DIPEA), 2997 Dipalmitoyl phosphatidylcholine liposomes, crospovidone, 3554
Diisoxazolylnaphthoquinone, chemical 1141 dissolution times, 3565
stability of, 390 nebulization, 3859 in hard gelatin capsules, 3565
Diketopiperazine (DKP) derivative, weight DipentumÕ capsules, 1254 interface-controlled mechanism, 3560
fractions, 4112–4113 Diphenhydramine HCL (DPH), 2014, 2423 rework efficiency (%RE) of, 3564
Dilatant flow, 3132 in tablet, 547 on swelling force kinetics, 3564
Dilated cardiomyopathy, in heart failure, Diphenylhydantoic acid, esters of, 943 sodium starch glycolate, 3553
1156 Diphosphoglycerate, 354 in tablet formulations, 3558

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I37

Disintegrants Dissolved organics and chlorine DNA–dendrimer complex, cellular

action, 3555 absorption of, 4040 metabolism of, 884
for tablet, 3556 carbon filters, disadvantage of, 4040 DNA–DNA reassociation values, 3526
swelling, 3556 organic contaminants, 4040 DNA-cationic vesicle, molecular architecture
water wicking, 3555 Distearoylphosphatidylethanolamine of, 1065
activity, factors affecting, 3560 (DSPE), 1333 DNA-damaging agents, 1413
disintegration time of Distillation Dobutamine hydrochloride, solubility
compression force on, 3561 definition of, 4042 enhancement in, 2124
surface response of, 3562 pharmaceutical water purification system, Docetaxel, 3360
molecular structure, 3561 4042 clinical development, phases of, 2812
particles Distortionless Enhancement by Polarization Docosahexaenoic acid (DHA), 2439
deformation recovery theory of, 3557 Transfer (DEPT), 3445 deficiency, 2439
repulsion theory of, 3557 Disulfide bonds, cleavage of, 1136 for mental health, 2439
in insoluble matrices, 3562, 3558 DitabÕ source, 2439
in soluble matrices, 3558 disintegration of, compression pressure on, Dodecanoic acid, in polyanhydride
in spray drying, 3230 3358–3359 preparation, 184
starches, 3230 Dithiothreitol (DTT), in mucus, 1347 Dodecanol, 2127
swelling of, 3561 Diuretics Dodecyl-N,N-dimethylamino acetate
in tablets, 3230 and digoxin, hyperkalemia, 1395 (DDAA), 1314
Disintegrating agents drug preparations, 703 Dodecyl-N,N-dimethylamino isopropionate
in tablet formulations, 3553 and lithium, sodium depletion, 1395 (DDAIP, NexACT 88)
mechanism action of, 3661 for diabetic patient, 1394 chemical structure, 1313
starch, 3661 hyperuricemic effect, 1394 skin permeation enhancer, 1313
tablets, 3661, 3662 and thiazides, 1394 Dodecyltrimethylammonium bromide, 1056
Disintegration test, 218 Diurnal rhythm, 1287, 3822 Dodecyltrimethylammonium chloride, 1066
Diskhaler, 1923 Divinylbenzene (DVB) hydrogels, 2023, Doehlert design, 2459
Dislocations in crystals 2025 in three dimensions, 2460
edge dislocation, 3545 Division of Drug Marketing Advertising and Dogs
screw dislocation, 3545 Communications (DDMAC), 63 drugs in, 3949
Disodium cromoglicinate (DNCG), 2107 DNA half-life in, 3962, 3963
as mast-cell stabilizer, 1123 chip, 1367 naproxen in, 3963
for asthma treatment, 1123 fabrication of, 1368 phenobarbital in, 3963
powder, 1123 parallel gene expression, 1368 systemic availability, 3949
Disodium cromoglycate, absorption of, degradation, 3912 Dolgit Mikrogel, 1126
1245 double-stranded, 930 Domoso (Syntex Veterinary Labs), 1887
Dispersed phase and the aqueous medium, electron spin resonance signal for, 3549 Donnan exclusion, 2121, 2126
mechanisms, 4117 endonuclease on, 930 DonnatalÕ, 3349
Dispersion systems genome-scale, 1367 Donor blood transfusions, 348
aqueous solution medium, 4117 immunization, 2751 Donor formulation, ionic strength of, 2749
electric double layer, 4117 microarray, 1367 Dopamine, 2148
dividision, 4117 fabrication of, 1368 agonists, 1417
Gouy–Chapman layer, 4117 parallel gene expression, 1368 Doppler shift
Stern layer, 4117 organic molecules interaction of measure of, 2588
neutral bulk-solution phase, 4117 acridine orange (AO), 936–937 monitoring of, 2589
pharmaceutical dosage forms, 4117 chromamycin A3, 936–937 Doramectin
phase, 4117 mithramycin, 937–937 topical solution of, 3971
immiscible liquid droplets, 4117 with phosphate backbone, 930 kinetic parameters for, 3955
solid particles, 4117 polymerase, 930 Dorsal-axial-inferior (DAI) lead system,
solid, 83 probes, 929 1411
as drug carriers, 4117 definition, 930 Dorsal dermis, 3821
Dispersive infrared spectrometers, 3409 growth measure, 929 Dosage form, 164, 988
double-beam, 3409–3410 in microbe identification, 929 acute conditions, 1104
optical null principle of, 3410 replication, 1161 advantages in, 419
single-beam, 3409 single-stranded, 931 adverse effects, 3768
Dispersive mixing, 2356–2357 technology aerosols, 988, 997
Displacement transducers, tablet press, 3666 construction of, 1133 in animals, 171
Disseminated diseases, 1140 recombinant, 260 application of, 2664
Dissociation constant KD, 3028 triplet code of, 931 bioavailability studies, 168
Dissolution vaccines, 3912 bioequivalence evaluations, 164
parameter, 1861 advantages, 3912 biowaivers for, 167–168
studies of, 2943 CpG, 3922 blood level studies, 168–170
testing, strength of, 2850 intraoral jet-injection technique for capsule, 164
Dissolution rate-limited absorption behavior, delivery of, 3916 chewable, 1610
2922 parenteral vaccination, 3916 chitosan-based, 999

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I38 Index

[Dosage form] [Dosage form] Double-beam spectrophotometers

chronic conditions, 1104 coating process, 1105 noise in, 3468
classification complexation, 1106 resolution of, 3468
by mechanism of release, 991 development, 1105 specifications of, 3467–3468
by type of formulation, 991 direct compression, 1107 types
clinical efficacy for, 3190 disintegration time in, 1108 double-beam-in-space system, 3465
coacervation, 1107 freeze drying, 1107 double-beam-in time system, 3465
coloring systems for, 664–667 granulation, 1105, 1106 Double-cone blender, 2355
controlled release oral, 2407 layering process, 1105 radioopaque grains in, blending of, 2367
creation and manufacture of, 948 manufacturing processes, 1105 grain concentrations of, 2362
delayed release, 991 molding, 1107, 1108 mixing patterns in, 2359
applications, 992 packaging selection, 1108 segregation evaluation in, 2363
coating materials of, 992 spray drying, 1105. 1106 Double-emulsion solvent removal method,
depends on pH values, 172 taste masking, 1105 2320
design, 2666 parenteral, 1616, 3954 Double-junction reference electrode, 1505
dispersed formulations, 3190 advantages and disadvantages, 3954 Double layer capacitor, 1494
dispersed particles, 3190 in animals, 3954 Double prodrug, 1256
disposition, 164 formulations, 3955 Double-stranded DNA (dsDNA)
drug-delivery systems, 3768 pediatric and geriatric, 998 base pair in, 931
drug release characteristics of, 991 pellet type controlled release, 991 definition, 930
elixir, 164 pharmaceutical, 648 sense strand of, 931
enteric-coated, 991 physical properties, 3190, 3257 Double-stranded RNA (dsRNA), 3147
extended release (ER), 991 plasma concentration, 174 in cytoplasm, 3148
factors affecting bioavailability of, 165 powder, 164 Douches, 955
factors preparation methods, 948 Dowcill 200, 3270
life cycle management, 1105 liquid type, 948 Doxercalciferol, 3347
patient preference, 1105 solid type, 948 Doxil, tumor accumulation of, 1332
formulation components, 3768 rapid dissolving technology (RDTs), 998 Doxorubicin, 675, 1184
hot-melt extruded, 2007 semisolid, 3257 conjugate of, 1329
immediate release oral, 2406–2407 semisolids, 988, 996 cytotoxicity of, 1192
immediate-release single-dose studies, 173 conjugate, 1139
in vitro dissolution testing, 171 solid oral, 3190 liposomes, 1142
gastric fluid, 172 dissolution study for, 3191 in vivo efficacy of, 1141
intestinal fluid, 172 solid type methods plasma levels of, 1192
in vitro evaluation of, 2850 medicated particulate solids, 948, 950 quenching of, 3391
by injection molding, 2007 medicated solid applications, 948, 950 Doxorubicin-loaded nanospheres
lipid excipients, 975 nonoral individual dosage forms, antitumor efficacy of, 1191
lipid, 982 948, 950 in liver metastases model, 1192
liposomal, 984 oral individual dosage forms, 948, 950 Doxycycline
liquid type method, 948 solid, 988 subgingival delivery of, 904
alcoholic or ethereal solutions and solution, 164 hyclate, 3360
preparations, 948–949 stereoisomerism implications, 3966 succinate, 2423
aqueous solutions and preparations, suspension, 164 Dragendorff reagent, to detect alkaloids,
948–949 systemic circulation, 3768 542
nonaqueous solutions and preparations, tablet, 164 Dreiding and Kendrew models, 715
948–949 therapeutic advantages, 3768 Dressings
oleaginous solutions and preparations, topical, 2408–2409 medicated, 955
948–949 topical, 988 paraffin, 955
parenteral solutions and preparations, type of, 3979 protective, 955
948–949 urinary excretion studies, 170 Dried material layer
sweet or viscid aqueous solutions, vaginal, 1339 vapour via
948–949 veterinary, 3941 migration of, 1439
liquid, 988, 993–996 Dosator machines, capsules, 410 solvent transfer of, 1439
liquid, advantages and disadvantages of, 28 Dose-dependent systemic hypoglycemia, 2743 Driers
manufacturer of, 988 Dose inhalers metered, 997 agitated batch, 3892
modified release (MR), 991 Dose toxicity studies, duration of, 124 batch, 3891
monographs on, 2831 Dosimeters circulation, 3892
multiple-dose studies, advantages of, 173 types continuous, 3893
for nasal route, 988 film badges, 3092 drum driers, 3893
non-paranterals, 988 pocket dosimeters, 3092 spray driers, 3893
oral, capsules and tablets, 28 thermoluminescent detectors (TLDs), fluid-bed, 3892
orally disintegrating tablets (ODTs), 1104 3092 hot air ovens, advantage, 3891
advantages, 1104 Double-beam-in-time spectrophotometer, solids, 3891
beads preparation, 1105 3466 spray, 3891

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I39

[Driers] [Drug(s)] [Drug(s)]

tumbling, 3892 assay methods compact mechanical strength of, 3643
vacuum tray, 3892 anticoagulant interference, 206 complexation of, 676, 677
Dronabinol, 3347 antioxidants, 206 by CDs, 680
Droplet residence time, 2083 assessment of, 206 complex biologic matrices, 194
Droplet size bacterial contamination, 206 components of, 1006
aerosol, 3856 cost of, 206 compound, chemical stability of, 3300
of nebulized aerosols, 3857 hemolysis, 206 concentrartion, 168
Dropping mercury electrode (DME), 1490 multicenter quality control schemes, 206 accuracy and reproducibility, 3484
Drops precision and accuracy, 206 in blood specimens, 3485
ear, 955 quantitation, 206 course of, 2802
eye, 955 sensitivity, 206 fluctuations in, 3484
nose, 955 beads in, 4068 by statistical distribution, 3485
toothache, 955 biliary clearance, 2154 containers
Dropwise condensation, heat transfer, 3873 binding, methods for, 3028 closure systems, 1477, 1480
Drude equation, 447 bioavailability of, 1013, 1075 categories in, 2528
Drug(s), 447 pharmaceutical factors affecting, 218–219 inhalation, 2545
absorption, 165, 209, 1228, 2152–2153 bioavailability, efficacy and safety, ADME types of, 2528
acid-catalyzed degradation for, 23 factors affect, 926 contamination control in, 2171–2172
advantage of bolus IV injection, 19 biologic material, 170, 1781 coprecipitation of, 3569
bioavailability, 19 in biological fluids, 516 corneal level of, 1196
buccal partitioning model of, 1075 biological variability, 3485 cosmetic, 798, 1782
characteristics of, 1305, 2868 biopharmaceutical properties of, 939, 941 cosolvency process in, 993
definition of, 212 biosynthesis of, 228 cosolvents in, 3323
differences in, 3823 methods, 228 ethanol, 993
absorption, distribution, and elimination significance of, 228 ethyl ether, 993
kinetics of, 2803 biotransformation, 2154 glycerol, 993
activity coefficient methods, octanol=water biotransformation and removal, 310 isopropyl alcohol, 993
partition coefficient, 3312 blood plasma concentration, 1214 isopropyl myristate, 993
addiction, 1038 brain delivery of, 2681 isopropyl palmitate, 993
brain disease, 1039 buccal absorption of, membrane storage in, propylene glycol, 993
characterization, 1039 1073 sorbitol, 993
administration route of, 32 buccal administration xylene, 993
advantages, 1071 advantages of and disadvantage of, 1079 covalent crosslinking of, 3049
anatomic site for, 1071 absorption kinetics of, 1073 crystalline, polymorphic forms of, 89
buccal cavity for, 1071 buffers in, 3317–3318 crystal properties, 3642
duration of, 2642 application of, 3317 crystals, 1729
mechanical system for, 2642 bulk densities of, 3643 decomposition of, 400, 403
method of, 3978 cancer treatment, 2469 definitions and nomenclature, 2527–2528
oral, 2544 capsule formulation for, 1672 degradation of, 165, 400, 1613
parenteral, 2544 carboxyl groups of, 1135 decomposition, 1272
pharmaceutical preparations for, cardiovascular diseases, 1783 in gastrointestinal tract, 165
types of, 1071 cardiovascular, 1075 hydrolysis, 1272
adsorption of, 35 carrier-mediated transport of kinetics of, oxidation, 1272
mechanism of, 40 1802–1803 pathways, 2859
aerosol carriers, for lymphatic targeting, 1611 primary pathways of, 1272
delivery efficiency, 2098 case study, 4031–4034 products, 2859
generation from, 2098 ligand-based versus Kv1.5 potassium rates of, 1612
particle size distribution, 2098 channel, 4020–4023 delivery vehicles for, 1062
airborne contaminants of, 2171 cellular metabolism of, 1301 design
analyses cephalosporin antibiotics, 170 chemical diversity, 1363
Bratton–Marshall approach, 195 characteristics, by supercritical processes, classical methods for, 1363
detection steps for, 194 3569 closures, 2532–2533
differential solvent extraction systems for, chemical basis for, 194 combinatorial chemistry, 1364
194 chemical instability, in biological media, computer-generated structures, 714
human=animal tissues and fluids, 3571 310 lipophilicity incorporated by, 1245
pharmaceutical preparations, 3572 chemical properties of, 19 methodology, 1363
separation method for, 194 chewable preparations of, 1076 multiple-unit closures, 2533–2534
anticholinergic effects, 1393 class variables, 3495 multiple-unit containers, 2533, 2540
anticoagulant, 72 clinical evaluation of, 560–570 rational, 1362
antimalarial, 1610 color of, 3642 single-unit closures, 2535–2536
apparent solubility of, 3313 colorants, 648 single-unit containers, 2534–2535, 2541
aqueous insoluble, 983 in Europe, 661–662 structure-based, 1362
aqueous solubility of, 939, 3483 compactibility of, 3643 development program, 2469

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I40 Index

[Drug(s)] [Drug(s)] [Drug(s)]

development, 1782, 1955 metabolism and pharmacokinetic data in, ingredients, 802
diffusion 1422–1424 versus cosmetic ingredients, 800
and drug release rate, 182 origins, 1406 inhalation
prediction of, 1886 phototoxicity testing in, 1418–1419 animal models, 21
discovery studies, 1406 containers, 2545
and development, 714, 2490 excipient, 1612, 1780 injectable intraocular delivery systems,
outsourcing, 2490 extemporaneous compounding of, 162
time and cost profile for, 2490 1009–1010 injectable
process, paradigm shift in, 2993 extractables and leachables in, 1693 cyclosporine, 983
disharmony methods, 1956 extraction ratio of, 1075 dissolution of, 983
dispersions, formation of, 766 extrusion, 1107 emulsion, 1004
dissolution rate of, 3313 factors compatibility, 1477 miconazole, 983
dissolution test first-pass metabolism in, 1424 paclitaxel, 983
assay technique, 916 food, 1782 suspension, 1006
basket apparatus method, 910 formation of, 761–762 teniposide, 983
cylinder apparatus, 914 melt granulation of, 762 instability
data presentation and interpretation, 917 formulation, 27 sources for, 2320–2321
delayed-release (enteric-coated) tablets design, 3645 strategies, 2321
and capsules, 922–924 of pharmacokinetic information in, intestinal mucosal membrane, 32
factors affect, 917–921 588–589 intolerance, mild reaction, 46
fiber optic technology, 916 of white film coating for tablet of model, intracellular delivery of, 1141
flow-through cell apparatus method, 1683 intracellular penetration of, 1189
912–913 formulation, metastable crystal in, 3313 intranasal application of, 1201
of flunixin meglumine, 924 free enzyme-labeled, 203 intravenous injection solution formulation
of immediate-release tablets and capsules, gastric emptying, effect in, 1423 for, 1670
922 gemtuzumab, 268 intrinsic dissolution rate (IDR) of, 3644
in vitro–in vivo correlation (IVIVC), 925 as generally recognized as safe and effective ionizable, solubility of, 3318
intrinsic dissolution method, 914–915 (GRASE), 2420 ionization of, degree of, 1074
paddle apparatus method, 911–912 glass, testing of, 2543–2544 ionized concentration of, 3314
paddle-over-disk apparatus method, 914 chemical resistance, 2543 label and package insert informations
principle, 910 light resistance, 2544 adverse reactions, 2504
procedures, 910–915 light transmission, 2543 contraindications of drug, 2504
reciprocating cylinder apparatus method, habituation categories, 1039 dosage and administration specifications,
912 hallucinogenic, 1040, 1045–1048 2504
reciprocating holder apparatus, 914 hard gelatin capsule formulation for, indications and usage, 2504
robotics and automation, 925 1671 overdosage effects and treatment, 2504
sample collection and analysis, 915–916 harmonization, 1958 pharmacology and biological action, 2504
of selected dosage forms, 922–925 heat-sensitive, 1475 substance and product and formulation,
versus drug release test, 922 high-extraction-ratio, 588 2504
distribution, 2153–2154, 2547–2548 history, 1001–1002 supply and storage conditions, 2504
volume, 2634 hydrolysis, 2040 warnings and precautions, 2504
docking algorithms, 4025–4027 hydrolytic degradation reactions labeling, 2504, 2546–2547
random or stochastic methods, 4027 carbonyl derivatives, 2040 layering process, disadvantage, 1106
dosage form, 1693 carboxylic acid derivatives, 2040 life-threatening illnesses, 1781
needleless injection, 1009 nucleophilic displacements, 2040 cancer, 1781
dosage form phosphoric acid derivatives, 2040 HIV, 1781
tool for, 2062 hydrophilic encapsulation methods, 4074 ligand designs, 705
type of, 2064 hydrophobic, 672, 1429 lignocaine hydrochloride, 1061
dose-effect relationship for, 573 hypotensive effects, 1395 lipid solubility of, 1074, 1245
efficacy, 1782, 2469 ideal solubility of, 3311 lipids to, chemical coupling of, 982
electroretinography in, 1411 identity and purity test, 3642 lipid-soluble-diffusible form of, 1074
elimination rate constant, 1013, 1014 imitanib, 268 lipophilicity of, 213, 543, 1225
enteric-coated dosage forms, limitations of, impact on, 2973 liquid, 307
942 impurity test, 3642 making of, 2905–2906
equilibrium solubility of, 225 in interference-free blood, 3483, 3485 manufacturing areas
evaluation in plasma specimens, 3497 Europe, 1958
antigenic sensitization in, 1418 by analysis of variance (ANOVA), 3492 Japan, 1958
carcinogenicity for, 1416–1418 from bioavailability study, 3495 United States, 1958
data application, 1406 in vivo in, 167 marketing, critics of, 57
definition, 1406 in vivo rate of, 2681 mathematical description of, 2758
immunotoxicity testing in, 1420–1421 in vivo–in vitro correlation (IVIVC) for, measure of, 2806
in vitro toxicity studies, 1419 3714 mechanism of, 1242
influence, 1407 induction times of parent, 843 metabolic transformation, 19

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I41

[Drug(s)] [Drug(s)] [Drug(s)]

metabolism, 169, 194, 1398 particle size, 3598 permeability, 2540
metabolite extraction, 194 physicotechnical properties of, 829 physicochemical, 2536
metabolizing enzymes, polymorphic, 587 and shape of, 3643 polymorphic behavior of, 771
metastable solid phase, solubility of, partition coefficient of, 1305 prescription, 1393
3314 passive diffusion of, 1079 polymorphic forms, formation of, 632
microencapsulation methods for, 2320 patient administration route of, 1693 postprocessing, 4030-4031
molecular structure of, 212 performance-enhancing, 1038 precipitation of, elimination of, 3317
molecular weight of, 1074 permeability class, 167 preclinical=clinical failure of, 2193, 2195
molecule, physicochemical characteristics permeability drugs, 3714 presystemic metabolism of, 1257
of, 672 peroral adminstration of, 1071 process of, 567
molten carnauba wax, 4074 pH control of, 3314 product
in molten excipients, 401 pH of saliva, 21 bioequivalent, 222
motility and dilution of, 1307 pH solubilized, 3317 design, 208–211
from mouth, pH on, 1074 pharamokinetic proerties of, 2816, 223 examples of, 224
mucosal delivery of, 1064 in pharmaceutical development, 1693 generic, 223
multiple oral doses of, 1013 pharmaceutical formulation of, properties
multipulse systems release of, 1287 disadvantages of, 1076 photochemical and photophysical, 2862
mutagenicity role in, 1413 pharmacodynamic properties, 1392 physicochemical, 2859
nonpolar cosolvent solubilization curves pharmacokinetic performance of, 425, 1392 protein structures, preparation of,
for, 3317 pharmacologic activity profile, 19 4023–4025
non-prescription, 1393, 2413 pharmacologic effect, 19 protein-binding sites, 1398
advantages of, 1393 arrhythmia, 1411 protein-bound fraction of, 1398
sedative effect in, 1393 central nervous system (CNS) proteins, 3029
non-specific oxidation of, 312 stimulation, 1411 psychedelic, 1048
chemical classes, 312 hypertension, 1411 psychoactive, 1038
non-steroidal anti-inflammatory pharmacophore mapping, 4018–4020 psychological modulation of bitterness,
ibuprofen, 3828 pharmacophore model, 4020 1106–1107
salicylates, 3828 pharmacophore searches, 4018–4020 quality attributes, 1782
normal distribution theory, 3488 strategies of, 4020 quality standards, 1958
ocular retention of, 1195 pharmacy compounding, 1789 random errors in, 3483–3484
opioid=analgesic, 1040 phenolic residues rapid absorption of, 2678
optical isomer of, 3436 conjugation to, 982 rapid precipitation of parent, 843
optimizing of, 1306 phenomenon of, 19 rates of, 4022
oral administration of, 1071 photochemical reactions, 2859 rectal absorption, 22
oral mucosa to photodecomposition, 2859–2864 reduction of, 1305
permeability of, 1072 photodegradation, 2859 regulation, 1781
organic, crystal density of, 3601 photolabile, 2862 at Food and Drug Administration
orpan photoprotection, 2862 (FDA), 1782
act’s, 2471–2472 photoreactivity of, 2859–2861 release, 4068
categories of, 2468, 2469, 2472 chemical functions, 2860 with bovine serum albumin (BSA), 4070
current status of, 2472 photosensitive, 2862 lactose, 4075
law, 2468 photostability pH on, 4069
pricing for, 2473 influence of product formulation on, renal clearance, 2154
survey, 2470–2471 2861–2862 renal diseases, 1783
osmality of, 291 studies, 2862–2864 repackaging, 2545
over-the-counter (OTC), 1780 test, 2859, 2862 conditions for pharmacists, 2546
oxidative chemical degradation of, 1967 physical and chemical stability of, 401 reproductive toxicology, 1414
package insert information of, 2504 physical properties of, 19, 2972 fertility studies (Segment-I studies),
packagers and repackagers, 2546 physical state of, 3297 1414–1415
packaging systems, guidelines and physicochemical and biopharmaceutical, perinatal and postnatal study, Segment-
regulations for, 2526 941 III studies), 1416
packaging, 1006–1008 physicochemical characteristics of, teratology studies (Segment-II studies),
parenteral delivery routes, 20 1073–1074, 1290, 1298 1415–1416
intra-arterial drug administration, 20 plasma captopril concentration of, 1074 as safe and effective (RASE), 2420
intra-arterial injection, 20 plasma protein binding of, 3034 safety, 1782
intramuscular (IM) administration, 20 plastics, testing for, 2536–2542 saliva, 160
intranasal administration, 20 agar diffusion, 2539 salts of, 3177
intrathecal injection, 20 biological, 2539 scoring functions, 4028–4030
intravenous (IV) administration, 20 elastomeric closures for injection, empirical type, 4029
transdermal delivery systems, 20 2538–2539 force field-based, 4028–4029
parenteral, 3997 in vitro, 2539 knowledge-based, 4030
advantages and disadvantages of, 1002 in vivo, 2540 sedative–hypnotic, 1040
particle characteristics, 3643 light resistance, 2542 serum levels of, 1073

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I42 Index

[Drug(s)] [Drug(s)] [Drug biotransformation]

size reduction of, 1997 polymers, 1106 classification factors, 321
solid dispersion of, 3592 polyvinyl acetate, 1106 environmental, 321
solid phase of, 3313 spray drying, 1106 physiologic, 321
solubility ratios for, 3314–3315 theophylline, 170 complex array of metabolites, 311
solid-state chemistry of, 3306 therapeutic products, 1779 cytochrome P450 in, 312
solubility and permeability, 2932 therapeutic serum concentrations of, diet constituents, 322
solubility of, 3311 1071 diet factors, 322
estimation of, 3312 therapy, continuous, 575 environmental factors, chemical exposure,
melting point on, 3312 considerations, 2424–2426 322
salt formation on, 3316 risks, 2426 enzyme systems, cytochromes P450 (CYP),
solubility, 213, 3318 thermodynamic activity of, 1306 321
cosolvents on, 3318 third class of, 2423 excretion route of
multiple cosolvents on, 3321 toxic effect study, 1408 metabolites, 323
and permeability groups, 3644, 3714 acute toxicity, 1408–1413 systemic excretion, 323
for polar solutes, 3320 chronic toxicity, 1410–1413 factors affecting, 321
for semipolar solutes, 3320 dosage selection in, 1412 genetic factors, 321
rate of, 3643 subacute toxicity, 1410–1413 hepatic (biliary), in glucuronic acid
solubilization of, degree of, 3320 subchronic toxicity, 1410–1413 conjugates, 323
solubilization, 3311 vehicle selection in, 1423 impact on routes of excretion, 322
by complexation, 3331 transcellular route of, 2674 in vitro method
by ionization, 3315 transport mechanism for, 1073 of biologic systems, 323–324
by pH control, 3318 trastuzumab, 268 collection and analysis of biologic
on cosolvent properties, 3320 unbound fraction of, 3029 samples, 323
on solute properties, 3320 un-ionized form of, 1074 qualitative information on excretion,
pH and complexation on, 3330 unionized form of, solubility of, 3318 323
solution formulation for, 3311 use and abuse levels quantitative data of, 323
stability and systemic availability of, 27 abstinence, 1038 subcellular preparation, 323
stability of, 308, 400 addiction, 1038–1039 isolated cells, 324
improvements in, 681 drug abuse, 1038–1039 of metabolites
parameters, 3715 experimentation, 1038 hepatic (biliary), 323
in solubilized systems, 3588 habituation, 1038 renal (urinary), 323
testing of, 2502 social=recreational, 1038–1039 physicochemical properties of, 323
statistical principles, 3483 variability, reduction of, 3814 monoamine oxidases in, 312
stereochemistry of, 3178 virtual screening, 4013 perfused organs, 324
stereoisomeric action of, 2149 database filtering, 4014–4017 phase reactions of, 311
eudismic analysis, 2149–2150 ligand-based, 4017–4023 physiologic factors
stereoselectivity in, 2150, 2152–2153 steps in, 4014 age, 321
steroidal anti-inflammatory, 3828 structure-based, 4023–4034 cardiovascular disease, 321
dexamethasone, 3828 volume of distribution of, 578 hormones, 321
hydrocortisone, 3828 water solubility of, 764 pharmacogenetics, 321
stimulant, 1040, 1043–1045 waterborne contaminants in, 2171 sex (gender), 322
storage, 2547–2548 water-insoluble, 3335,983 in pregnancy, 321
sublingual and buccal dosing of, advantage in water-soluble matrix, 1107 renal (urinary), in sulfate conjugates, 323
of, 1076 water-soluble, bioavailability of, limitation stress factors, 322
surface activity of, 3595 of, 774 Drug-carrier systems, properties of, 645
surface area of, 3643 wax granules in, 4068 Drug and cosmetics (D&C) red, 657, 664
surfactants in, 31 wax matrix, 4071 pharmaceutical, 4049
in suspensions, 28 wetting agents, 3648 and drug products, 3726
suspensions, physical stability of, 831 with biodegradable material coat, 1235 and dosage forms
syringe pump for, 2642 Z-statistic for, 3488 thermal analysis techniques, 3726
systematic errors in, 3483 advertising and promotion of, 57 aging, 3742
systemic circulation, 19 Drug–antibody amorphous state, 3733, 3735
tablet formulation for, 1667 complex, 1570 applications, 3733
in tablet matrix, 1107 conjugate, 1140 combined techniques, 3731
taste masking of, 1105 decreased, 1480 differential scanning calorimetry (DSC),
complexation, 1106 ratios, 1140 3726
ethylcellulose, 1106 Drug biotransformation differential thermal analysis (DTA),
Eudragit E 100, 1106 alcohol and aldehyde dehydrogenases in, 3726
granulation, 1106 312 Gibbs phase rule, 3733
hydroxypropyl cellulose (HPC), 1106 animal species hot stage microscopy combination,
hydroxypropyl methylcellulose (HPMC), in drug development, 322 3731
1106 in safety evaluation, 322 kinetic study transformations, 3738
incorporation of, 1107 cDNA-expressed human enzymes, 324 microcalorimetry, 3729

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I43

[Drug and cosmetics (D&C) red] [Drug-delivery systems] [Drug-delivery systems]

phase diagrams, 3733 efficiency, 2124 by nebulizers, 988
polymorphism, 3733 enzyme-activated, 1099 sandwich-type, 1087
preformulation, 3742 equipment, 3722 self-regulating, 1100
principles, 3726 excipients in, 3641 shapes and sizes of, 1083
processing, 3742 factors influencing, 179 site-targeting, 1082, 1101
pseudopolymorphism, 3733 feedback-regulated, 1082, 1099 sonophoresis-activated, 1096
thermogravimetic analysis (TGA), formulation, 3719 targeted, 1149, 1328
3726 gastrointestinal route of, 1777 techniques for, 179, 999, 1082
thermomechanical analysis (TMA), hydration-activated, 1096 transdermal therapeutic system,
3726 hydrodynamic pressure-activated, 1094 3973–3974
Drugs and cosmetics (D&C) and external hydrogels as, 2032 transdermal, 924–925, 1313
D&C colarants hydrolysis-activated, 1099 vaginal route of, 1177, 1399
dyes, 656 hydrolytically labile polyesters in, 182 vapor pressure-activated, 1094
lakes, 656 intravenous, 1010 variability of, 2866
solubilities and fastness properties of, 668 ion-activated, 1098 Drug delivery technologies, 2105
specific uses and restrictions of, 656 iontophoresis-activated drug, 1096 application of, 1104
Drug–coarse carrier interaction, 1430 liquid crystals in applications of, 1123 devolopment of, 1248
Drug delivery devices to lungs, 2077, 2093 features, 2106
inhalation magnetic-activated, 1095 pharmaceutical market, 1104
dry powder inhalers (DPIs), 2094 marketing tools, 1104 oral transmucosal technologies, 1105
metered dose inhalers(MDIs), 2094 matrix system for preparation of, 181 pH-specific dosage forms, 1105
nebulizers, 2094 mechanical force-activated, 1095 physical and chemical properties, 1104
performance comparison of, 2116 method of, 3979 Drug delivery vehicles, characterization,
pneumatic nebulizer, 2093 microreservoir partition-controlled, 1087 1054
pressurized metered dose inhaler (pMDI), miscellaneous, 2409 Drug-dendrimer complex, 1064
2093 monoclonal antibodies for, 1143 Drug-depletion zone, 2670
squeezing bulb nebulizers, 2093 mucoadhesive hydrogel, 1169 Drug-design methodology, 1363
Drug-delivery systems mucosal routes for, 1169 Drug development, 714
activation-modulated, 1082, 1090 nasal route of, 1175 chemical structures, 1901
administration of, 295, 1169 needle-free, 1209 cycle, 2826
injection, 295 neutron scattering studies of, 1054 drug design, 1901
oral, 295–296 ocular route of, 1176 drug–protein complexes, 1902
pulmonary, 296 from oil=phosphatide emulsions, 984 effective global planning, 568
topica, 296 ophthalmic solutions, 1220 food-effect assessment in, 2821
advantages of, 1082 oral colon-specific, 1228 inhibitory potency, 1902
aerosol, 2081 orally disintegrating tablets (ODTs), 1104 life cycle of, 2553
afficionados, 2753 taste masking priciples, 1105 and manufacturing, 109
amorphous, 87 osmotic pressure-activated, 1090 optimization of, 1902
applications osmotic pump, 992 phase 1 to phase 4, 560, 563
polyesters for, 179 parenteral emulsions for, 984 phases, 3022
polymer properties versus performance, particulate, 1331 physicians in role of, 561–562
180 of patient compliance, 1082 preclinical and clinical testing phases of,
biodegradable, synthetic and natural pH-activated, 1098 2193
polymers in, 179 pharmacological effects of, 1141 process, 92, 560, 2802
bioerosion-regulated, 1099 polycaprolactone (PCL), 1223 departmental interactions, 2, 1371
bioresponsive, 1100 polyesters in examples of, 182 departmental responsibilities, 1,
buccal route of, 1174 polyglycolide (PGA), 1223 1370
chronotropic, 999 polylactide (PLA), 1223 regulatory affairs works, 2, 1371
from collagen shields, 1223 polymer hybrid-type, 1087 review of, 1, 1370
from contact lens materials, 1221 polymer matrix diffusion-controlled, successful management of, 3, 1372
control excipients, 999 1085 program, 560, 2824, 2922, 3004
controlled-release, 1169 polymer matrix system in, diffusion- protein binding pocket, 1902
degradation and resorption of, 177 controlled release rate of, 182 R&D ‘’pipeline’’ in, 2194
in dermatological therapy, 1317 polymer membrane permeation-controlled, randomization process in, 566
dermatological, 1313 1082 research, goal of, 570
design of, 1245, 1258, 1332 polymer selection criteria for, 180 roles of, 561
device, 2665 polyorthoesters in, 186 simulation in, 2807
devices, 296–297 polyphosphoesters in, 189 techniques, 258, 260
disadvantages in needle use, 1209 problems in, new polymeric materials for three Rs (3Rs) in, 2192
distribution pattern of, 1138 solving, 179 Drug–disease interactions, 1392
drug molecules, 1082 rate of, 3814 Drug–drug interaction, 171
drug reservoir gradient-controlled, 1087 to respiratory tract, 988 complex formation by, 701
effects of, 2640 by inhalation aerosols, 988 pharmacodynamic interactions, 1911

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I44 Index

[Drug–drug interaction] Drug-metabolizing enzymes, induction of, [Drug packaging materials]

pharmacokinetic interactions, 1911 1018 polyethylene (PE) containers, 2529
studies, 563 Drug-microcrystalline cellulose mixture, polyethylene terephthalate G (PETG)
Drug–excipient interactions, 388, 1613–1614, X-ray powder diffractometry (XRD) containers, 2529, 2531, 2542
4113 pattern of, 4107 polyvinylidene chloride (PVdC), 2531
amorphous product, 4113 Drug–nutrient interactions, 51 types of, 2528–2532
crystalline, 4413 Drug package, 562 composition
X-ray powder diffractometry (XRD) batch mixture for amber soda-lime glass, 2528
technique for, 4113 container, 2511 properties
Drug Efficacy Safety Implementation (DESI) batching and melting, quality control in, types, 2528
programe, 1787, 2419 2511–2512 and suitable application of, 2529
Drug Export Act, 1780 blow-molding and tubing process, Drug partition coefficient, 578
Drug extraction and analysis, modes of, comparison of, 2517–2518 Drug product, 1580, 1622, 3928
3571 bulk powder, 2085 administration of, 3483
Drug–food interactions, 1911 composition, 2508 aging problems, 3747
Drug–herbal interactions, 52 composition, properties, and classification, analysis, 3747, 3797
Drug-in-adhesive type, 2925 2513–2515 animal-derived materials, 1641
Drug-induced hepatotoxicity, 1420 container aseptic manufacture principle of, 2290
Drug information systems, 1385–1390, 2855 aluminum cans, 1971 benefits and risks of, 59–60
compendia, 1385–1386 blow-molding process, 2513 bioavailability, 164, 166, 3483, 3711
formularies, 1386 characteristics, 2517–2519 analytical method, 174
internet resources, 1389–1390 chemical durability, definition by ASTM, assessment of, 171
BioMedNet, 1388 2518 design of, 172
criteria=issues for selecting, 1390 classification of, 2521 determination of, 168
MedMaster, 1389 Danner process, 2513 factors affecting, 164–165
pharmweb, 1388, 1389 process of, 2512–2514 formulation, 165
pharmacopoeias, 1385 tubing process, 2513 manufacturing variables, 165
primary sources, 1388–1389 type I, II, III, 2521 physiologic factors affecting, 165
e-journals, 1388–1389 definition by bioequivalence (BE) studies, 3711
patent information, 1388 American Heritage dictionary, 2508 analysis data, 174
secondary sources American Society for Testing and analysis of variance (ANOVA) method,
American Chemical Society (CAS), Materials (ASTM), 2508 174
1386 delivery, 2085 crossover design for, 172
biosciences information service (BIOSIS) energy sources for, 2086 plasma concentration, 169
previews, 1386 formulation:batch mixing, charging, and statistical methods, 174
CAplus file, 1386–1387 melting, 2509–2510 biopolymers, 1643
criteria=issues for selecting, 1387–1388 manufacture composition, 2508–2509, buffer, 1627
EMBASE, 1386 2510 chelating agents, 1625
International Pharmaceutical Abstracts material, glass, 2508 classification of, 3797–3798
(IPA), 1386 product, quality of, 2510–2511 gas chromatography=mass spectrometry,
MEDLINE, 1386, 1388 properties of, 2515–2517 3799–3801
pharm-line, 1387 thermal expansion, 2516–2517 liquid chromatography=mass
Drug Master File (DMF), 1401, 1472, 1632, tests, 2521–2524 spectrometry, 3801–3805
1617 apparatus used for, 2522 compatibility of excipient with, 1638
advantages of, 1405 arsenic, 2524 containers
drug application, 1402 light transmission, 2524 and closures, 3271
format requirements, 1402 powdered, 2522 control of, 1944
packaging, 1404 reagents for, 2522 control of, 3798–3799
process, 1402 treatment cost-effectiveness claim for, 59
regulations of, 1402 interior surface, 2519–2520 decapeptylÕ, 1643
Drug interactions printing, 2519 dermatologic vasoconstriction, 171
consequences of, 1392 as unit-dose, 2084 devolopment of, 778
determination of, 570 Drug packaging materials dissolution profile of, 909
development of, 1394 containers, types of, 2529–2531 expiration dating period, 1685
enzyme induction, 1398 metal, 2531 expiration intervals, determination, 1685
factors contribution of, 1392 paper extemporaneous preparation of, 2643
non-prescribed drugs, 1393 and cardboard, 2532 film-coated, 1735
pharmacological effects, 1392 paperboard formulation, 1622
prescribed drugs, 1393 plastic, 2529 special additives, 1630
inspection of, 565 characteristics for, 2530 homeopathic, 1687
mechanisms of, 1393 polyolefins, 2529 human gastrointestinal tract, 171
reducing risk of, 1399 polystyrenes (PSs), 2529 identification of, MS-based systematic
of therapeutic agents, 1395 polyvinyls, 2529 approach for, 3809–3811
Drug-liposome interaction, 4127 polyvinyl chloride (PVC) containers, 2531 in vitro dissolution testing, 172

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I45

[Drug product] [Drug products, biotechnology-derived] Drug–PSA polymer interaction, 2931

in vitro surrogate tests of, 3271 instability, pysical, 282–283 Drug–receptor
in vivo release assessment, 3710 stability-indicating profile, 302 complex, 2802
in vivo=in vitro correlation (IVIVC) for, stability testing, 302 drug interactions, electrophysiological
168 Drug products, formulation development, analysis of, 3116
injectable, 1622 287–295 electrostatic interactions, 722
inorganic, 3797 excipient selection, 289 flow cytometric methods, 3116
inspection, 1946 antioxidants, 292 fluorescent formyl peptide ligand, relatime
labeling of, 59 buffer and pH, 290 analysis of, 3116
Lupron DepotÕ, 1643 preservatives, 292–293 formyl peptide receptor, 3116
lyophilized as, 1623 salts, sugars, and polyols, 291 green fluorescent protein (GFP), 3117
manufacturing process, 3928 surfactants, 291–292 human beta-adrenergic receptor, 3116
marketing, 1622 state selection of, 287–289 interactions, 714, 3116
in Japan, 1622 stability studies, accelerated, 294–295 ligand–receptor interactions, analysis of,
in United States, 1622 Drug release 3116
microbial contamination of, 2790 anitumor, 1777 molecular interactions, 716
microbiological contamination of, 2286 control molecular pharmacological techniques,
microbiological standards for, 2286 advantages and disadvantage of, 29 3116
microorganisms, 1590 biological half-life, 30 nicotinic acetylcholine receptor (LGCR),
microscopic examination of, 3273 characteristics of, 29 3116
MS-based analytical techniques, 3799–3809 development of, 29 protein kinase activity, 3116
multiple dose administration, 173 dosage forms, 29 quantitative analysis of, 3116
nutropin DepotÕ, 1643 by microencapsulation, 2322 theory schematic representation of, 2803
parenteral, 1622 oral, 30 Drug–resin complex, 1106
non-official excipients, 1632 particle size influence on, 2323 Drug solubility, 808, 3319
percentage of excipients to, 1638 polymer chemistry on, 2322–2323 complexation in
pharmacokinetic perturbations affecting, and device erosion, 180 inclusion, 3328
166 dissolution rates of, 909 stacking, 3327
powder as, 1623 estimate of, 924 cosolvency on, 3320
quality checking and Fickian diffusion, 181 mathematical description for, 3319
chemical analyses, 3707–3709 factors affecting, examples of, 180 effect of, 1306
dissolution test, 3712 of high molecular weight drugs, by potential effects on, 817
pharmaceutical availability, 3709 formation of pores or channels, PSA polymers, 2930
quarantine of, 1946 182 Drug solubilization
residual solvents, 3797 low-molecular-weight drugs effect of, 182 advantages of, 2914
safety and efficacy assessment of generic, low-molecular-weight drugs vs. high capacity, 1059
925 molecular weight drugs, 182 cosolvents
single dose administration, 173 methacrylate and polyvinylpyrrolidone definition, 2914
stability test, 1687, 1477, 3272-3273 copolymers in, 1221 drug solubility, 2914
ICH guidelines, 1685, 1686 mucoadhesive devices of, 2035 water-miscible organic solvents, 2914
protocols for, 1687 polymer degradation rate and mechanism emulsifiers, 2914
steam sterilization, 2291 of, 180 emulsion, 2914
structure process for, 3809 profiles, 1288 limitations of, 2914
tablet press instrumentation for, R&D rate of, 1308 methods for, 2914
grade instrumentation, 3689 drug diffusion on effect of, 182 solubility properties, 2914
trace level impurity in polymer degradation on effect of, 182 surfactants, 2914
identification of, 3799 factors affecting, 181 Drug substance
International Conference on recent regulatory initiatives, 1320 crystal form, 3002
Harmonization (ICH) guidances for, retardants, 2004 aqueous solubility, 3002
3797–3798 retarding effect in, 1288 bioavailability, 3002
organic, 3797 roll surface configuration on, 3167 biological characterstics of, 3002
scope of, 3797–3799 sustained formulations chemical characterstics of, 3002
zoladexÕ, 1643 liquid, 1129–1130 crystallinity, 3002
Drug products, biotechnology-derived semisolid, 1129solid, 1129 dissolution rate, 3002
degradation=inactivation, 282–283 test, 2850 physical charcterstics of, 3002
filling, finishing, and packaging, 306–308 triggers, 1327 relative toxicity, 3002
instability, chemical, 283–287 Drug–polymer crystalline racemates, 3740
deamidation and succinimide formation, complex, 3028 Schröder–Van Laar equation, 3741
283–284 dissociation constant of, 3033 crystallinity, isothermal microcalorimetry,
disulfide exchange, 286 interaction 3738
fragmentation, 287 and drug stability, 178 crystallization of, 3004
glycation, 287 of ionised groups on, 180 differential scanning calorimetry (DSC)
isomerization=racemization, 286–287 study of, 3028 curve
oxidation, 284 mixture, 40, 2670 magnesium stearate, 3745

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I46 Index

[Drug substance] Dry-heat sterilization, 3530 [Dry powder inhalation (DPI)]

mannitol, 3745 advantage and disadvantage of, 3517 devices
differential scanning calorimetry (DSC) applications of, 3060 and powder behavior, 2087
purity analysis, 3739 complicating factors of, 3517 blends and homogeneous powders in,
dissolution rates of, 909 from Compton effect, 3900 2086
DSC scan of, 398 humidity of, 3515 bulk powder drug package, 2085
enantiomers, 3740 in pharmaceutical industry, 3512 design objectives and constraints, 2085
interaction with water, 3747 kinetics of, 3512 functional description, 2085
pH stability profile of, 390 microbial death rate curves, 3513 performance specifications, 2087
phase diagrams, 3743 microorganisms and endotoxins by, dosage form, 2078
polymorph, 3742 destruction of, 3512 drug containment categories
polyvinylpyrrolidones (PVP) in, 3745 mixing of liquids, 3900 reservoir, 2078
preformulation, 3742 and moist heat sterilization, 3512 unit dose, 2078
purity determination from photoelectric effect, 3900 formulation, 2081
direct method, 3741 processes, types, 3513 filling for, 2084
indirect method, 3741 sterile parenteral products manufacture of, in crystalline state, 2079
Prigogine–Defay equation, 3741 2291 packaging for, 2084
purity of, 398 variables physical state effect on stability in, 2079
solid solution, 3745 microbial water content, 3516 manufacture, 2081
solubility, pH dependence of, 390–391 open and closed systems, 3516 microparticulate powders of, 2082
stabilization of, 389–390 temperature, 3516 characterization, 2086–2087
TG, 3738 Dry-heat temperatures, 3513 specifications, 2087
with excipients, 3737, 3745 Dry-heat tunnels pharmaceutical operations
Drug validation and ovens, 3514 lyophilization, 2078
analytica procedures, 92 sterilization milling, 2078
factors affecting, 92 exposure time, 3514 spray drying, 2078
types of, 94 in pharmaceutical industry, 3513–3514 product quality requirements, 2088
as good business practice, 93 Dry milling respirable powders of, 2079
methods selection, 92 air attrition=fluid energy, 2347 system characteristics, 2078
and method variation, 93 electrostatic agglomeration, 2345 therapeutic applications, 2078
method and product specifications, 93 equipments for, 2345 battery operated, 1541
Drum blenders, 2355 flow ability, 2345 oral inhalation, 1540
axial demixing in, 2358–2359 impact mills, 2345 powder dispersion energy, 1541
competitive patterned demixing in, 2359 media (ball) milling, 2348 Dry powdered inhalers (DPIs)
mixing patterns in, 2357 particle size distribution, 2345 advantages of, 1283
radial demixing in, 2358 thermal degradation, 2345 for asthmatics, 1284
segregation pattern in, 2358 Dry powder breath activated, 1283
Drum driers, 3893 factors for manufacture, 2080 dry powder aerosols, 1283
components, 3893 for inhalation, 2077 macromolecules for, 1283
costs of, 3893 packaging systems type I alveolar cells, 1283
spray and splash feeds in, 3893 reservoir systems, 2085 Dry seal cachets. See Cachets.
Dry agglomerates, 2258 unit-dose systems, 2084 Dry solid filling capsules, 411
Dry blend process aerosol formation, 2085 Dry dosage regimens, design of, 584
cohesive drug product, 3206 Dry powder inhalation (DPI), 34, 2089 Drying
low-dose drug product with, 3205 adhesion forces, 3249 desorption of water, 1817–1820
lubricant mixing time, 3206 advantages and disadvantages, 2078 leaning equipment process steps, 1582
uniformity effects on, 3205 in asthma therapy, 3248 mass transfer resistance, 1812–1813
Dry eye syndromes, 1220 carrier-containing formulation, 3249, 3250 moisture content during storage, changes
Dry granulation process, 3663 chlorofluorocarbon propellants in, 1819–1820
compression stage, 3661 ozone-killing effect of, 3248 optimization of, 1808
conventional tablet press, 3661 cohesion forces, 3249 optimum residual moisture, 1817–1818
roller compactor tablet press, 3661 formulation principles, 3248, 3250 process control
on interparticulate bond formation, disodium cromoglycate, 3250 and chamber pressure, 1815–1816
3161 relative humidity, 3250 end point, determination of, 1816–1817
lubricant, 3662 gravitational forces, 3249 condenser performance, effect of, 1816
roller compaction, 3159 micronized particles, 3249 product temperature measurement,
slugging, 3159 narrow particle size, 3250 1813–1815
tablet manufacturing process, 3661 of secondary electron microscopy, product temperature, 1812–1813
Dry-heat batch sterilizer, 3514 3253 product temperature control and glass
in pharmaceutical industry, 3513 stabilizing mechanism, 3249 transitions, 1817
Dry heat belt sterilization, 748 excipients types rate of, 1812–1813
Dry heat depyrogenation, 3512 glucose, 3250 secondary, 1817–1820
Dry-heat destruction, of microorganisms with lactose, 3250 stabilization during, 1827
soil, 3516 and metered-dose inhaler, 2090 Drying air flow rate control, 1445

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I47

Drying applications Ear canal, external [Effervescent products]

of pharmaceuticals, 1436 antiseptic agents for, 2477 physiological reactions, 1457
Drying curve, of spherical particle, 1438 epithelial layer of, 2476 charcoal preparation, 1457
Drying process, 2083 inflammatory condition of, 2475 direct compression
constant rate period of, 1443 isoelectric point of, 2475 compressible mixture, 1458
falling rate portion of, 1443 pH level, 2475 free-flowing, 1458
first falling rate period of, 1443 predisposing factors, 2476 non-segregating, 1458
in pharmaceutical materials, 1435 skin of, 2475 of raw materials, 1458
phases of, 1443 EasimistÕ, 3855 drug compositions, 1457
second falling rate period of, 1443 Easson–Stedman’s hypothesis, 2149 dry granulation
from solids, 1443 of stereoisomeric drug action, 2150 roller compaction, 1458
Drying rate, of spherical particle, 1438 E-BRITEÕ alloy, 792 by slugging, 1458
Drying system, with flow characteristics, 1440 Echinacea, effects of, 69 environmental factors, 1458
DSC. See Differential scanning calorimetry. Echinacea angustifolia, 71–72 fusion granulation method, 1458
Disperse phase volume, 1052 Echinacea pallida, 71–72 paracetamol (acetaminophen), 1457
Dual-chamber syringe delivery system Echinacea purpurea, 71–72 processing equipment, 1458
advantage, 3917 Eclectus. See Linctus. tableting, 1458
cost of, 3917 Eclegma. See Lohoch. wet granulation methods
vaccine, 3916 Econazol, antimycotic, 1127 infrared (IR) spectrophotometry, 1458
Dual-electrode detector, 1522 Ecosom Liposomengel. See Pevaryl Lipogel. X-ray diffractometry, 1458
Dual wavelength reflection scanning EcotrinÕ, 1254 Effervescent reactions
for pharmaceutical analysis, 543 Ectoparasiticides classes, cattle, 3972 acid–base reactions1454
Dulbecco’ phosphate-buffered saline, 2130 Ectoparasitic preparation, 3981 acidity for, 1455
Dulcolax, 983 Electrical distribution system, 1489 pharmaceutical dosage forms, 1457
Duodenal irritation, possibility of, 942 Electronic document management system vacuum processing, 1458
DuragesicÕ, 1084, 2129 (EDMS) Toolbox, 2558 Effervescent salts. See Salts, effervescent.
DuraSolv technology, tablets manufactured Efavirenz, dose of, 3349 Effervescent stability
with, 1110 Effervescent dosage forms compression pressure, 1461
Durham–Humphrey Act, 1787 advantages and disadvantages of, 1456 compositions for, 1461
Durham-Humphrey Amendment, 2419 oral solid dosage forms, 1456 monitoring
Durometer hardness, rubber, 1474 Effervescent excipients, 1290 cantilever beam proximity transducer,
DUROSÕ, 3360 Effervescent formulation 1461
Dust explosion, 2346 antiadherents, 1459 mercury-intrusion porosimetry, 1461
Dyes antifoaming agents, 1459 with effervescent products, 1460
as colarants, 664–669 binders, 1459 Effervescent tablets, 990
properties and applications, 668–669 diluents, 1460 acid materials, sources of
shade and stability of, 667 disintegrants=dissolution aids, 1460 acetylsalicylic acid (aspirin), 1456
solubility and stability properties of, excipients, 1459 acid anhydrides, 1456
667–668 flavors, 1469 acid salts, 1456
Dynal# magnetic beads, 932 glidants, 1459 ascorbic acid, 1455
Dynamic differential scanning calorimetry hydrogenated maltodextrins, 1460 citric acid, 1455
(DDSC), 394–396 lubricants, 1459 fumaric acid, 1455
heat flow measurement maltitol, 1460 malic acid, 1456
heat capacity component of, 395 optimization, 1460 tartaric acid, 1455
kinetic component of, 395 solid dispersions, 1460 biopharmaceutical aspects, 1457
Dynamic fluorescence quenching, 1972 surfactants, 1460 carbon dioxide, sources of
Dynamic laser light scattering, 305 sweeteners, 1460 amino acid–alkali metal carbonate
Dynamic light scattering, 639, 1058, 2386, water-soluble colors, 1460 derivatives, 1456
2589 Effervescent granulation, production of, calcium carbonate, 1456
Dynamic mechanical analysis (DMA), 3731 746 potassium bicarbonate (KHCO3),
for polymers, 3731 Effervescent granules, 1454 1456
pharmaceutical applications, 3731 air suspension coating–reacting technique, potassium carbonate, 1456
Dynamic oscillatory techniques, 1881 1462 sodiumbicarbonate (NaHCO3), 1456
Dynamic spray devices tableting, 1462 sodium carbonate, 1456
clean-out-of-place (COP) method, 1583 Effervescent ibuprofen sodium glycine carbonate, 1456
Dynamic vapor sorption instrument (DVS), bioavailability of, 1457 compression, 1454
for water sorption–desorption plasma concentration curves, 1458 in process tests, 1462
isotherms, 3730 tablet, 1457 monitoring of, 1462
Dyslipidemias, treatment of, 72 Effervescent products, 1454, 2978 dissolution, 1454
acetylsalicylic acid (aspirin), 1457 dosage forms, 1456
Ear antacid preparations, 1457 drugs (product categories), 1457
anatomy and physiology of, 2475 aqueous solubility, 1455 evaluations
drops, 994 biopharmaceutical aspects chemical properties, 1462
xylene in, 993 gastrointestinal tract, 1457 non-destructive method for, 1462

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I48 Index

[Effervescent tablets] Electrical sensing zone technique, 1796 Electrochemical sensor

physical properties, 1462 Electric circuit, charge carrier in, 2747 definition, 1506
granules, 1463 Electric double layer stoichiometric equivalence point, 3753
Karl Fischer analysis in, 1462 counter-ions in, 4119 Electrochemical transducer, 1522, 2058
for oral solution, 1454 definition, 640 Electrochemiluminescence, 1576
packaging, 1463 division Electrodeless discharge lamp (EDL), 3367,
aluminum-foil blisters, 1463 Gouy–Chapman layer, 4117 3395
dye-solution test, 1463 Stern layer, 4117 Electrodes
physical deterioration of, 1463 electrostatic repulsive force, 4121 cell-and tissue-based, 1526
plastic materials, 1463 formation of, 1254, 1493 cells, tubular packed-bed, 1522
plastic tubes, 1463 of pharmaceutical particles, 4117 enzyme-based, 1526
physical stability, 1463 Electric potential formulations, pH in, 2748
powders for, 1463 inner, 1502 for ion migration, 2121
raw materials, adsorption=desorption gradient, impose of, 2747 in iontophoretic systems, 2121
isotherm, 1454 Electroactive materials, 1507 medical potentiometric, 2121
Effervescent vaginal suppositories, 1457 Electroactive organic compounds, 1490 mercury, 1521
Efflux pump Electroactive species packed-bed, 1521
definition of, 2719 cathodic reduction of, 1490 platinum, 1521
inhibitors, 2720 diffusion coefficient of, 1493, 1497 polarization, 1491
Efflux transporter P-glycoprotein, 2720 Electro-acoustic effects, dynamic mobility processes, mechanism of, 1493
Egg yolk phosphatidylcholine (EYPC), determination, 4119 Electrode-solution interface, diffusion at,
fornmation of, 1059 Electroanalytical method, 1516 1495
Eicosapentaenoic acid (EPA), 2439 Electrocapillary curves, 1490 Electrogenerated oxidizing agents, bromine,
Ejection cam tablet press instumentation, Electrochemical biosensor, structure of, 3765
3690 1525 Electrohydrodynamic (EHD) device
Ejection force transducer, lubricants Electrochemical cell, 1502 by Battelle Pulmonary Therapeutics, Inc.
evaluation, 3692 for potentiometry, 1516, 1517 (BPT), 2114
Elan’s NanoCrystalTM technology, 2384 Electrochemical detection, 1516 dose uniformity of, 2115
Elasticity, 1466 applications, oxidative, 1528–1530 formulation, 2114
loss of, 1467 for pharmaceutical analysis, 1516 in vitro and in vivo performance, 2114
Elastic moduli, 1291 off-line techniques, 1524 lung deposition efficiency, 2115
Elastomers, 793, 1466, 2242 principle for, 1499 monodispersed aerosols by, 2114
chemical structures of, 1469 Electrochemical detector, 1521, 534–535 particle size distribution of a NCE from,
fluorinated, 2242 by absorption spectroscopy, 202 2114
limitations of, 794 amperometric circuit, 202 process, 2114
materials biomedical trace analysis, 199 pulmonary drug delivery devices, 2114
chloroprene, 2275 diffusion current, 202 scintigraphic images of, 2115
ethylene–propylene diene monomer direct current (DC) pulse polarograph, soft aerosol cloud delivery by, 2114
(EPDM), 2275 202 Electrokinetic capillary instrumentation, 204
peroxide-cured acrylonitrile=butadiene, mobile phase for, 202 Electrokinetic sonic amplitude (ESA), 4119
2275 pharmacologically active compounds, 202 Electrolysis experiment, 1491
thermoplastic elastomers (TPE), 2275 in reversed phase liquid chromatography, Electrolytes, 1809
in parenteral packaging components, 202 chemical nature of, 3779
1468 voltametric oxidation, 202 concentrations, 1395
pharmaceutical and chemical applications voltametric reduction, 202 of colligative properties, 3779
of, 794 voltamogram analysis, 202 freezing-point depression, 3779
physical and chemical, 1478 Electrochemical enzyme immunoassays, interionic interactions, 3779
saturated and unsaturated, 1467, 1470 homogeneous, 1528 tri-univalent, 3779
synthetic, 1466 Electrochemical flow cell, 1576 uni-univalent, 3779
Elastomeric dressing, 1032 design of, 1521–1523 Electrolytic deposition, 1498
Electrical power sources, 1482–1483 Electrochemical immunoassay, 1526–1528 Electromagnetic deflection system
Electrical power systems, 1482 advantages of, 2059 components of, 3219
feeders and branch circuit arrangements, conductometric detection, 2058 distortion-free images, 3219
1486–1488 electrochemical detection, 2058 Electromagnetic radiation, 1050, 3084–3085,
generators and emergency power sources, immobilization of, 2058 3540
1488 potentiometric detection, 2058 Electromagnetic spectrum, 3405, 3461
grounding, 1489 Electrochemical methods, 1490 absorption band in, 448
harmonics, triplet, 1488 applications, 1971 radiation types in, 3406
primary configurations, 1486 comparison, 1974–1975 region of, 3434
primary electric service arrangements, in headspace oxygen anlysis, 1970 Electromigration, 2119–2120
1483 operation principle, 1970–1971 Electron beam sterilization, 749
secondary configurations, 1483–1485 Electrochemical potential, definition of, Electron capture, 3085
utility services, 1483 1502 Electron capture detectors (ECD), 466
Electrical resistance, 1303 Electrochemical reaction, 1491 Electron density, delocalization of, 697

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I49

Electron-donor groups [Electrophoresis] Electuaries, 955

ligands, 3760 light-scattering method Elettaria cardamomum, 1764
dissociation of, 3391 electrophoretic velocities determination Eleutherococcus senticosus (Siberian ginseng),
Electron energy loss spectroscopy (EELS), of, 4119 73
2397 zeta potential, determination of, Elixirs, 955, 994, 3349
Electroneutrality, of atom, 3082 4119 of sulfanilamide, 2904
Electronic absorption bands, 449 Doppler shifts, 4119 Emax model
spectra, 447 motility, 830 drug–receptor occupancy theory, 2803
spectrum, 3387 Nernst-Planck theory, principle of, 3847 non-linear concentration–effect
Electronic batch record (EBR) system, therapeutic compound, 3847 relationship, 2803
10, 741 Electrophysiologic chareacterization, 2671 Embrocatio. See Liniments.
Electronic communication, 4101 Electroporation, 2119–2120 EmcocelÕ, 3675
Electronic document management applications, 2749 EmcompressÕ tablets, 3558
environment, 2556, 2553 deep electric current penetration, 3850 EmdexÕ powder, 3557
goals and objectives of, 2553 definition, 3847 centralized registration procedure, 1981
efficiency and effectiveness, 2553 intrinsic skin permeability, 3847 decentralized registration procedure, 1982
increase consistency of classification, mechanisms, 3849 Emetine, biosynthesis of, 245, 246
indexing, 2553 electroosmosis, 3849 EmisphereÕ technology, 999
power of, 2555 electropermeabilization, 3849 Emission spectrophotometry
sharing of documents, 2553 electrophoresis, 3849 for alkali earth metals analysis, 3373
Electronic excitation, 3541 membrane, permeabilization of, 3847 for metal analyses, 3367
Electronic integration, 474 physical impaact of, 2749 operating principles of, 3372
Electronic pulse counters, 639 on skin effect, 2750 Emission tomography, 3089
Electronic records skin permeation, physical methods of, Emissivity, of spherical particle, 1437
components of, 2562 1311 Emollient creams, as skin barriers, 1315
controls for, 2556 transdermal drug delivery, 3849 Empirical Hill equation, 2808
final rule on, 2556 Electrorepulsion and electroosmosis, Emplastrum. See Plasters.
guidelines on, 2551 mechanism of, 2746 Emulsification, 1551, 1996
rules on, 2560 Electrorepulsive contribution, 2747 ultrasonic process of, 3268
Electron ionization Electrosprays (ESI) Emulsification–evaporation processes, 3590
energetic electron beam, 1699 advantages of, 1544 solvent evaporation, for nanosphere
fragmentation, 1699 agricultural applications, 1544 preparation, 1187
spectrum for, 1700 atomization technique, 1543 techniques, 1560
Electron microscope components of, 1544 heat sterilization, 1561
Abbe’s equation, 3217 for dispersing suspensions, 1544 preparation, 1561
electron beam, 3217 droplet production zone of, 1544 Emulsifier, 995-996
principle of, 3217 drug molecules, 1544 anionic, 3261
refractive index, 3218 electrohydrodynamic sprays, 1543 cationic, 3261
resolving power of, 3217 electrostatic dispersion, 1544 classifications, 1551
types of, 3217 electrostatic sprays, 1543 in cream formulation, 3261
wavelength, 3218 heat-labile substances, 1544 finely divided solids, 1551
Electron paramagnetic resonance, 701 intense electric field, 1543 natural (macromolecular) polymers,
Electron spectroscopy for chemical analysis killing efficiency, 1545 1551
(ESCA), 2241 macromolecules, 1545 surface active agents, 1551
Electron spin resonance (ESR), for lipid monodisperse droplets, 1544 for semisolid emulsions, 3263
analysis, 981 for neutralization, 1544 high-turbulence, 3267
Electron transfer, 1492 respiratory delivery efficiency, 1544 hydrophilic–lipophilic balance (HLB) value
reaction, 1493 Electrostatic charge interactions, 3278 of, 3261
current, 1492 Electrostatic coulomb excitation, 3541 lecithin, 4075
Electrooptic modulator, 450 Electrostatic density, 716 monoglycerides, 4075
Electroosmosis, 2119–2120 Electrostatic interactions, 40, 697 multiagitator, 3267
effect of, 3847 Electrostatic potential, 716 natural polymeric, 1558
electrokinetic phenomena, 3847 Electrostatic powder deposition non-ionic, 3261
with electrophoresis, 3848 coating process, 1539 pharmaceutical, 1551
pH dependent, 3847 image charge force, 1539 phase behaviour of, 1559
Electroosmotic convective flow, 2747 oral dosage forms, 1539 surfactant, 1552
Electropermeabilization space charge force, 1539 triethanolamine stearate, 3261
bilayer composition, 3849 Electrostatic properties, 3056 types, 3261
flux enhancing mechanism, 3849 Electrostatic sterilization, of pharmaceutical Emulsifying agents, 1275, 1551, 1624
pore formation rate, 3849 products, 1545 cremophor EL, 1624
spontaneous formation, 3849 Electrostatic technology finely divided solids, 1553
Electrophilic reactant, 437 electrospraying from natural products, 1553
Electrophoresis, 304 for respiratory delivery, 1535 interfacial tension of, 1275
ionic species, movement of, 3847 pharmaceutical sterilization, 1535 polysorbate, 1624

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I50 Index

[Emulsifying agents] [Emulsion] [Enantioseparation]

preservatives for in water in-oil emulsions, 1184 cyclodextrins in, 2158
advantages of, 1553 preservatives in, 996 gas chromatographic (GC) procedures,
applications of, 1553 preparation of, 3267 2156
sodium desoxycholate, 1624 pseudoplasticity and thixotropy of, 995 liquid chromatographic (LC) procedures,
Emulsifying devices, 1561 radiopaque, 1550 2156
Emulsion rheological properties of, 3143 ligand-exchange process, 2158
advantages, 1548 control, 995 mobile-phase-additive technique for,
characteristics continuous phase, 1554 2157
conductivity measurements, 1554 particle size distribution, 1554 Enantiotropy, 2936
droplet size distributions, 1554 phase volume, 1554 polymorphs, 399
external phase, 1554 semisolid, 996 versus monotropy, 2936
rheology, 1554 stability Enatioselective enzymatic oxidation, 3002
classification, 1548 colloid theory, applications of, 1557 EncapsinTM, 684
composition, 956, 1548 classical theories of, 1557 Encapsulation
continuous phase, 1548 Derjaguin, Landau, Verwey and capsule dosage form, 3206
dermatological, 1548 Overbeek (DLVO) theory, 1557 capsule shells, 3206
diprivan, 984 electrostatic effects, 1557 efficiency, 1335
dizac, 984 stabilization mechanisms, 4122 equipment, 424
dosage form, 4122 surfactants in, 995–996 filling machines
droplets thermodynamically unstable, 1548 automatic, 3207
deposition of, 644 water-in-oil, 1548, 995 dosator-type machines, 3207
size of, 1187 ionization of emulsifying agent, 1557 dosing disk machines, 3207
surface of, 1052 polyether surfactants, 1557 principles of, 3206
drug bioavailability, 1549 solid particles, 1558 types of, 3206
in drug release, 1549 steric effects, 1557 formulation, advantages of, 3206
formation van der Waals forces, 1557 gelatin capsules, 3206
multiple type, 1554 surfactants in, 4123 instrumentation of, 3207
oil-in-water type, 1554 zeta potential application to, 4123–4124 Encephalitis vaccine, 4101
water-in-oil type, 1554 system Encephalopathy, 2633
formulations of, 1548 aqueous phase of, 3267 Encryption and certificate authorities, 2556
clarithromycin, 1549 plastic containers for, 3271 Endocrine-disrupting chemicals, studies of,
diazepam, 1549 Emulsion-base components, 3261 1950
globule size, 3269 Emulsion=hydrogel combination, 1128 Endocytosed proteins, 2724
heterogeneous preparation, 1548 Emulsum. See Emulsions. Endocytosis
for intravenous administration, of lipid Enantiomer, 2142, 3741 definition, 156, 1244
nutrients, 3589 application of, 3967 receptor-mediated, 27
ingredients in, 996 of amphetamine, 2143 cellular uptake of immunoglobulin, 27
injectable, 4123 bioavailability of, 3967 low density lipoprotein, 27
electrostatic stabilization of, 4123 biotransformation, 320 role of, 156
flocculation rate, 4123–4124 of chiral drugs, 3967 processes, 2718, 2672
pH effect on stability of, 4123 chromatographic separation of, 2156 Endocytozable formulation, 1190
physical instability of, 4123 dextrorotatory, 2143–2144 Endogenous and surrogate ligands, 3123
lipid, 1550 levorotatory acute antimigraine drugs, action of, 3123
manufacture of, 996 disopyramide, 2154 agonist effects, ananlysis of, 3123
manufacturing process, 349 drug stereoselectivity of, 2149 antimigraine drugs, 3123
mechanisms by, 1549 enantiomer interactions, 2155 Endogenous compounds
for medications delivery, 4122 in formulating dosage forms, 3967 chemical analytical methods, 194
nasal applications, 1550 gastrointestinal absorption of, 2150, conjugation reactions, 317
non-ionic surfactants in, 4123 2152–2153 glucuronidation, 317
ocular applications, 1550 ibuprofen, 2156 physical analytical methods, 194
oil-in-water, 995, 1548, 4122 low melting point, 3742 physical separation of, 194
oral, 1549 optical purity (enantiomeric purity) of, Endogenous inhibitors, accumulation of,
parenteral, 984 2144 3037
particle-electrolyte interaction pindolol, 2154 Endogenous pyrogens, 3054
non-specific adsorption, 4123 protein binding of, 3034 Endogenous substrates
specific adsorption, 4123 racemic compound, 3741 glucuronidation, 318
perfluorochemical, 1550 racemic drugs, 320 of methylation reactions, in detoxication,
pharmaceutical, 1996 resolution and racemization, 2144 320
applications, 1548 terbutaline, 2154 sulfation, 318
pharmacokinetic characteristics, 1550 Enantiomer–enantiomer interaction, 3034 Endometrial cancer cells, 71
polymerization, 861, 1064, 2333 Enantiomers=structural isomers, 3436 Endometriosis, treatment of, buserelin in,
in emulsifier free systems, 1184 Enantioseparation 1078
in oil-in-water emulsions, 1184 chiral stationary phases (CSP) in, 2159 Lupron DepotÕ for, 178

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I51

Endonuclease enzyme, 930 Environmental microbial ecology, changes in, Enzyme-linked immunoassays, 1524
Endopeptidases, 2717 3982 Enzyme-linked immunosorbent assay
Endorphins, 1040 Environmental monitoring (ELISA), 203, 1567, 2052, 3049
Endosomolytic polymers, 1063 data, 3070 competitive, 1568
Endothelium functions, 1326 media in, 2311 illustration of, 1567
Endotoxin programs, 595 technique, 1527
aggregated endotoxin vescicles, 3056 Environmental pollution, 438 Enzyme-mediated chemiluminescence
apolysaccharide structure, purification of, Environmental Protection Agency (EPA), immunoassays
3053 436, 1931, 1948 antigen–antibody reaction, 2058
bilayers break down of, 3056 Environmental scanning electron microscopy catalyst=cofactor, 2058
biological effects of, 3055 (ESEM), 2395 components of, 2058
chemical nature and structure of, 3053 Enzymatic barriers, 1611 Enzyme-multiplied immunoassay (EMIT),
contamination, 595 Enzymatic catalysis application of, 3001 203, 1528
depyrogenation, effective process for, 3055 Enzymatic cleavage, 1256 illustration of, 1567
destruction of, 3516 Enzymatic degradation, 2669 Enzyme immunoassays, 1526
disseminated intravascular coagulation, drugs susceptible to, 1301 advantages of, 2051
3054 nasal epithelium, 2683 application of, 2684
dry-heat temperatures of, 3517 peptides, degradation mechanism of, 2683 heterogeneous electrochemical, 1527, 1528
elimination of, 358 xenometabolic activity, 2683 homogeneous electrochemical, 1528
lethal effects of, 371 Enzymatic drug stability, 1303 induction
mammalian defense system, stimulation of, Enzymatic hydrolysis, 3008 enzymes for, 1398
3054 absence of, 3013 in excretion, 1398
physical properties of, 3055 of chemical bond, 1327 in metabolism, 1398
preparations of, 3053 desymmetrization, 3002 inhibitors, 942, 1611
pyrogens, 1267 Enzymatic production of RNAi libraries from coadministration of, 2725, 2728
reverse-osmosis membranes, 3056 cDNA (EPRIL), 3154 enzyme inhibitors, 2686
size of, 3056 Enzymatic reaction, 1524 protection of, 2686
sterilizing membrane filters, 1267 Enzyme proteolytic enzymes, species of, 2686
test for, 2830 alteplase, 272 spectroscopic properties, 2051
threshold pyrogenic doses of, 3055 biochemical molecules, 2204 vasopressin, nasal absorption of, 2686
tumor reduction, 3055 for catalyzing reactions, 2204 Eosinophilia–myalgia syndrome, 2447
Endotoxin alert limits (EAL), 3060 cleaners, of contact lenses, 2205 Ephedra sinica and nevadensis, 74
Endotoxin limit (EL), 3059 deficiency, in emphysema, 275 Ephedrine
fraction of, 3060 efficacy of, 1867 chiral centers of, 2145
bacterial, 2789 electrodes (Biosensors), 1525–1526 hypotension, treatment of, 4100
tests, 3060 antigen–antibody reaction, 1566 isomers for, 2145
Endotracheal absorption, 2633–2634 bioanalytical method, 1566 and pseudoephedrine stereoisomers, 2146
Endpoint analysis, 2555 classification, 1566, 2051 and pseudoephedrines, enantiomers in, 459
Endpoint chromogenic reactions, 3059 conditions for, 1566 (-)Ephedrinum 2-naphthalenesulfonate
Endpoint detection, in neutralization critical binding reagents, 1575 enantiomeric purity of, 398
titration, 3755 for low-molecular-weight analytes, EpiDermTM, 2197
Enemas, 994 1568 Epidermal cytochrome P450, 3970
liquid, injections of, 956 heterogeneous, 1566, 2051 Epidermal growth factor (EGF), 885
therapeutic substances, 956 homogeneous, 1566, 2051 Epigallocatechin gallate, 2438, 2444
Energetic electron beams, 3540 immunoassay, 1566 Epigastric distress, 1245
Energy dependent drug efflux pump, 1248 sensitivity and specificity of, 1578 Epigenetic carcinogenesis, mechanism of,
Enhanced permeability and retention (EPR) technological advancement, 1575 436, 437
effect, 884, 1326 validation of, 1572 Epigenetic carcinogens, 431
Enprofylline, synthesis of, 2999 proteins manipulation, 266 Epigenetic effects, 443
Enrofloxacin eliminatrion, microsomal- reactors, 1524–1525 Epinephrine, 944, 2862
mediated oxidative reactions, 3965 therapy for Gaucher’s disease, 2473 Epithelial absorptive cells, 2715
Ensete ventricosum, 3231 transferring Epithelial cancers, 1328
Enteric-coated systems (ECS), 1254 N-acetyltransferase, 3961 Epithelial cell
Enterocytes, 1177 methyltransferase, 3961 apical surfaces of, 1301
plasma membrane of, 577 sulfotransferase, 3961 cell membranes of, 1246
Enteroendocrine cells, 1177 UDP-glucuronyltransferase, 3961 dysplasia, 897
Enterprise Resource Planning(ERP), 2166 Enzyme-activated drug delivery systems, layer, 1301
Enthalpy fabrication of, 1099 apical membrane of, 1301
interaction, strength of, 602 Enzyme-based electrodes, 1526 polarization of, 2720
change, 394 Enzyme-catalyzed reactions, 1523 membrane, negative charged, 2726
fusion (DHF), 2943 Enzyme-controlled systems, 1254 Epithelial lining, 1072
Entropy Enzyme-labeled antigen, 1567 Epithelial lining fluid(ELF), 1280, 2737
repulsion effects, 1558 Enzyme-linked electrochemical techniques, alveolar epithelial cells for, 1281
omission of, 2941 1523–1526 surfactant, 1281

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I52 Index

Epithelial membrane [Equipment cleaning] EstradermÕ system, 984, 1084

surface, pH control at, 1302 cleaning solution, 1584, 1585 Estradiol cream, 1354
vesicle, 2723 concentration, 1585 Estradiol delivery, 1349
Epithelium cell layer, 2716 non-aqueous cleaning products, 1583 Estrogen
Epithema selection, 1584 metabolites of, 149
constituents, 957 water quality, 1584, 1585 receptor-a (ER-a), 2442
dry, 957 residue limits, 1587–1588 replacement therapy, 71, 1343
liquid, 957 riboflavin testing, 1583 vaginal creams, 1349
soft, 957 sampling procedures prooxidant effects of, 148
Epixodes direct surface sampling, 1588 Estrous cycles, seasonal breeding species,
hydration of, 316 placebo sampling, 1589 3943
hydrolase activity, in mammalian tissues, rinse sampling, 1589 Esystem data, titration curve, 3757
317 swab sampling, 1589 Ethacrynic acid, of drug release, from
Epoetin molecules, 270 soil condition, 1586 polyphosphazenes, 188
Epoxidation steps, 1581–1582 Ethambutol stereoismoers, 2146
aromatic double bond, 313 disassembly, 1581–1582 Ethanol, 1611, 3335, 3350
of double bonds, in unsaturated drying, 1582 and cremophor EL, 3361
hydrocarbons, 313 isolation, 1581–1582 cosolvent systems
hydrophobic molecules, 317 prewashing, 1582 regression analysis of, 3322
Epoxide rinsing, 1582 solubility for, dissolution rate with,
glutathione, 318 washing, 1582 3313
hydrolase activity validation, 1580, 1586 solubilization slope and solute polarity
cytosol of liver, 317 change control, 1591 for, 3321
endoplasmic reticulum of liver, 317 maintenance, 1590 in HFA MDI solution formulations
Epoxy resin coating, 2274 monitoring, 1590–1591 advantages, 2272
Equidae, 3948 Equitransferent salts, definition of, 1505 disadvantages, 2273
Equidimensional morphology, 824 Ergot alkaloids, biosynthesis of, 247 layer, frozen, 2318
Equilibrium dialysis (ED), 3027 Ergotomine, hallucinogenic effects of, 1046 metabolism, 703
Equilibrium dissociation constant, 3028 Erodible polymeric delivery systems, 1222 precipitation step, 3999
Equilibrium solubility, 85 Eroding suture materials, 1223 as sedative hypnotic drug, 1042
Equilibrium solution concentration, measure Erosion corrosion solvent for indicator solutions, 3755
of, 863 forms Ethanol-induced hypoglycemia, 2639
Equilibrium vapor pressure versus cavitation, 783 Ethanol–water composition, solute in,
temperature, of solvents, 1440 fretting, 783 solubility of, 3321
Equipment cleaning Erosion-controlled lag time, 1289 Ethical review processes (ERP), 2192
analytical method, target residue, 1589 Ery-TabÕ, 1254 Ethinylestradiol, 1350
baking, 1586 Erythema Ethyl- and methyl parabens, mixtures of,
characteristics degree of, 2671 1821
amount of soil, 1586 skin reaction characterized by, 1315 Ethyl alcohol in colloidons, 993
nature of equipment surfaces, 1586 Erythrocyte Ethyl cellulose, 156, 991, 2008, 2012
physical nature of soil, 1586 drugs in, 3033 and paraffin wax, 4069
construction materials, 1581 membranes, lipid peroxidation of, 141 Ethylenediamine (EDA) core
degradation product, 1587 vesicles, size of, 1335 amidoamine branching structure, 872
design characteristics, 1581 Erythromycin, 1399, 1419, 1441 size exclusion chromatography, 876
drying, 1586 coadministration of, 1257 viscometry, 876
factors, 1581 salts, 3184 Ethylenediamine tetra-acetic acid (EDTA),
degree of disassembly, 1581 Erythropoietin 292, 694, 3270, 3760
extent of automation, 1581 for needle delivery and needle-free delivery, as calcium salt, 1625
good manufacturing practices (GMPs), 1215 as injectable product, 1625
1580 pharmacokinetics, 1215 2-Ethylhexyl acrylate, copolymer of, 2928
methods plasma concentrations, 1215 Ethylene glycol
clean-in-place (CIP), 1581 Erythroxylum coca, 247 in diol pre-polymer preparation, 190
clean-out-of-place (COP), 1581 Escharotics, 957 reaction time, 3522
microbial control issues, 1590 Escherichia coli, 261, 1028, 1180, 3910, 3957 Ethylene glycol diglycidyl ether (EGDGE),
operational qualification (OQ), 1587 Esophageal cancer, 1254 2023
parameters Esophageal retention, 2867 Ethylene glycol dimethacrylate (EGDMA),
temperature, 1581, 1585 Esophageal transit, 2866, 3871 2022
time, 1581, 1584–1585 Ester structure, 2023
pharmaceutical manufacturing, 1580 hydrolysis reactions, 2040 swelling, 2025
procerdure for, 1943 of aspirin, 2043–2044 Ethylene oxide
processes, 1580 classification, 2041 advantage of, 1472
advantages, 1587 of haloperidol decanoate, 311 biological activity of, 3519
agent, 1581, 1583 phthalate, 1733 characteristics, 3519
aqueous cleaning products, 1583 sebacate, 1733 concentration, effects of, 3519

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I53

[Ethylene oxide] Eudragit RSPM, 2012–2013 [European union]

decomposition properties of, 3522 Eudragit S, 1234 concertation procedure, 1594
emission control, 3521 European Agency for the Evaluation of institutional partners, 1597
by catalytic oxidation, 3522 Medicinal Products (EMEA), 1594 legislation, 1593
by thermal incineration, 3522 arbitration, 1599 decisions, 1594
by water scrubling, 3522 cooperation with competent authorities, directives, 1594
emission limit of, 3521 1596 for pharmaceutical products, 1594
exposure, 3520 in European pharmacopoeia (EP), 1598 for veterinary products, 1594
emergency situations, 3520 management board, 1594 recommendations, 1594
maintenance operations, 3520 mission, public and animal health, 1594 regulations, 1593
sterilization equipment operation, 3520 principal scientific bodies, 1595 types of, 1593–1594
sterilized product degassing, 3520 referrals, 1599 legislative instruments, 1593
as fumigant, in food industry, 3519 scientific committees, 1595 medicinal regulatory authorities, 1597
gaseous alkylating agent, 3901 structure, 1594 member states, 1593
hazard assessment of, 3520 for synthetic drugs, 1596 multistate procedure, 1594
hot degassing of, 3521 units, 1595 European Union drug regulatory authorities
molecules, collision of, 3520 administration, 1595 network (EudraNet), 1597
occupational engineering considerations of, medicines for human use, evaluation of, European Union Guide to Good
3520 1595 Manufacturing Practice (EU GMP),
packaging of, 3520 medicines for veterinary use, evaluation 2134
physical and chemical properties of, 3520 of, 1595 European working standards
polymerizes in liquid state, 3901 technical coordination, 1595 congealing of, 763
sterilant mixture, 3522 working party, herbal medicinal products, establishment of, 2830
sterilization 1596 Eutectic mixtures, simple, preparation of,
biological indicators for, 3526 European Department for Quality of 775
control of, 3524–3527 Medicines (EDQM), 406, 1598 Evaluation of medicinal products (EMEA),
mechanism of, 3519 European Economic Area (EEA), 1594 1687
of medical devices, 3519 European Economic Community (EEC), Evaporation
vacuum pump modifications, 3521 powers, 1593 definition, 1600
validation of, 3524–3527 European Free Trade Association (EFTA), processes of, 1600
ventilation systems in, 3520 1961 rate of, 3004
toxicity of, 3901 European fruit brandies, 1042 Evaporative light scattering detector, 535
vapors of, 3519 European Medicines Evaluation Agency Evaporator
Ethylene oxide (EtO)-carbon dioxide mixture, (EMEA), 2197, 3071 advantage of, 1600, 1602
sterilization of, 3523 European monitoring centre for drugs and applications for, 1600
Ethylene oxide (EtO)-hydrochlorofluoro drug addiction (EMCDDA), 1596 classification, 3880
carbons (HCFCs) mixture, 3524 European Network of Official Medicines costs associated with, 1606
with sterilization cycle, 3525 Control Laboratories (OMCL), design of, 1602–1606
Ethylene–propylene diene monomer 1598 disadvantages of, 1602
(EPDM), 2275 European parliament, 1593 disk or cascade, 1600
Ethylene-propylenediene rubber, 2237 European pharmacopeia, 406, 1598, 1662, efficiency of, steam-heated, 1602
synthetic elastomers, 1466 2857 film evaporators, 3880
Ethylene–vinyl acetate polymers, 945, characteristics of, 2829 forced-circulation evaporators, 3880
1234 supranational document, 2832 natural circulation, 3880
Ethylenevinyl acetate compilers of, 2831 operation and control, 1607
copolymer fiber, 903 editions of, 2829 thermal stability of solution, 3879
membrane, 2932 goals of, 2829 types, 1600–1602
Ethylene vinyl alcohol (EVOH), 2933 legalities of, 2829 agitated film or wiped film, 1602
Ethylphosphodichloridate (EOP), in medicinal products, quality of, 2829 circulation, 1602
polyphosphoester preparation, prescription of, 2830 forced circulation, 1602
190 promotion of, public health, 2829 horizontal tube, 1602
Ethynodiol diacetate-releasing vaginal purpose of, 2829 jacketed, 1601
devices, 1346 role of, 2829 long-tube vertical, 1601
Etoposide, 3360 European quality award, 3077 plate-type, 1601
EudragitÕ, 991, 1234, 1736, 1797 European regulatory environment, 2 short-tube vertical, 1601
chemical structure, 1235 European regulatory guidance, 2772 unit process in pharmacy, 3879
water-soluble and water-insoluble, 1234 European Scientific Cooperative for Evening primrose oil (EPO) (Oenothera
Eudragit buccal patches, 2035 Phytomedicines (ESCOP), 2904 biennis), 72
Eudragit E-100 extruded films, mechanical European Technical Office for Medicinal Exacerbate infection, 371
properties of, 2014 Products (ETOMEP), 1597 ExcedrinÕ, 4106
Eudragit L, 1234, 2010 European union, 1593, 1632, 1656, 1789 Excentric tablet press, 3670
Eudragit L 30 D, 1737 history of, 1593 density distribution, 3654, 3665
Eudragit L–Eudragit S combinations, central and eastern European countries lower punch, 3667
1234 (CEEC), 1597 upper punch, 3667

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I54 Index

Exchangers [Excipient(s)] [Excipient(s)]

chloride–bicarbonate, 27 fine particle fraction, 1650 microencapsulation
sodium–calcium, 27 and formulation incompatibility encapsulation efficiency, 1652
sodium–hydrogen ion, 27 acid–base chemical reaction, 1652 of macromolecules, 1652
Excipient(s), 764–765, 1622, 1656, 3647 excipient crystallization, 1653 microsphere structures, 1652
active ingredients in, 41 excipient–drug interactions, 1653 nerve growth factor (NGF), 1652
additives, 1646 excipient-mediated water distribution, polyethylene glycols (PEG) excipients,
adverse effects of, 1641, 2776 1653 1652
aging of, precirol, 3746–3747 solid-state crystal transformation, 1653 surfactant excipients, 1652
amorphous, 1648, 1822 tableting excipient, 1652 moisture content of, 3643
animal model for testing, 1661 thermal analyses, 1653 mutagenicity testing, 1659
antioxidants, 1625 formulation, 1662 absorption, 1659
application, 1657 for oral administration, 3336 distribution, 1659
construction, 1658 in freeze-dried (lyophilized) powders excretion, 1659
cosmetics, 1658 process, 1646 metabolism, 1659
food, 1658 stabilizers for proteins, 1648 mycotoxin, 1641
paint thickeners, 1658 function, 1624, 2453 non-active ingredients, 1646
approval mechanics, 1658 functionality tests, 3642 oily injectable formulations, 3362
food additive petition, 1658 gas chromatography (GC) method, 1633 oral formulations, 3334
New Drug Application (NDA), 1658 Good Manufacturing Practices (GMP) for, microemulsion, 3348–3349
bioburden and endotoxin limits, 1641 3642 surfactants in, 3335
bulking agents, 1627, 1629 to gauge processing parameters, 3933 oral solutions and elixirs, 3349
capsule formulation, 415 harmonization, 1963–1964 osmotic agents, 1650
category of, 1624 hygroscopicity of, 38, 3643 particle size of, 3275
changes in impurities, 1614–1615, 1961, 1962 pharmaceutical, 84, 1622, 3938
bioburden, 1657 in hydrophile–lipophile balance values, cellulose derivatives used as, 4059
chemical properties, 1657 416 formulations, 3334
functionality, 1657 inferior grades of, 2777 starch derivatives used as, 4059
impurity profile, 1657 ingredients as, 1638 pharmacopeial standards for, 1962
moisture level, 1657 injectable formulations phospholipid-based, 1643
physical properties, 1657 aqueous-based solubilized, 3351–3354 physical properties of, 2779
characteristics, 2978 nonaqueous solubilized, 3355–3359 physical stability of, 1641
chelating agents, 1625 water-soluble organic solvents, 3350 polyethylene glycols (PEG), 3742
chemical shifts in, 3303 interaction with, 700 polymeric and surface active compounds,
chemical stability of, 1641 lactose, heat treatment, 3742 1623
classes, 3647 liquid, 3747 powder aerosol formulations
compatibility of, 1638 literature, 3647 pharmaceutical inhalation aerosols,
concentration, 1623 manufacturability, 3647 1648
controlled-released solid dosage forms, manufacturers, awareness of, 2779 treatment of diseases, 1648
polymeric excipients, 1652 materials powders, 3278
crystal morphology of, 828 celluloses, 4049 preservatives, 1625–1626
crystalline, 1648 gelatin capsules, 4049 antimicrobial, 1626
polymorphic forms of, 89 starches, 4049 benzyl alcohol, 1625
crystallization of, 1809–1810 measurement of, 2776 chlorocresol, 1626
definition, 1658 mechanisms of, 1646 efficacy of, 1627
degradation, 1641, 2453 melt congealing processes parabens, 1625
development, regulatory guidance for, bile acids, 764 process AIDS, 1613
2779 citric and succinic acids, 764 profile addresses, 1657
diethylene glycol, 1780 ethylacrylate copolymers, 765 properties, 37
dilution capacity of, 3646 glycerides and polyglycolyzed glycerides, bulk density, 34
drug carrier system, 1648, 1650 765 compactibility, 34
drug delivery system, 1658, 3641 hydrogenddated vegetable oils, 765 flowability, 34
in drug product, 1622 hydroxypropyl methylcellulose (HPMC), protectants, 1628, 1629
drug targeting, 1612 765 in protein pharmaceuticals
dry powder inhaler formulations, 1650 methylmethacrylate copolymers, 765 non-parenteral use feasibility, 1647
degree of crystallinity, 1650 natural and synthetic waxes, 765 parenteral use suitability, 1647
flowability, 1648 poly (ethylene oxide) (PEO) polymers, redox reaction potential, 1647
powder formulation, 1648 765 for qualitative optimization, 2453
effects of, 2453–2454 polyethylene glycols (PEG), 765 quality control, by glass transition, 3737
emulsifying agents, 1624 sterols, 764 reducing agents, 1626
ethoxylated, 1641 steryl esters, 764 regulatory process for, 690–691
evaluation criteria, 1657 sugars, 764 retrospective harmonization, 1955, 1959
FDA inactive ingredient guide, 1633 urea, 764 lactose, 1960
fermentation processes, 1641 metal complexing, 389 magnesium stearate, 1960

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I55

[Excipient(s)] Excitation and emission spectra, correction Extrusion, 745, 1712

route of use of the candidate of, 3401 applications in pharmaceutical industry,
eye irritation testing, 1659 Excitation grating, 3397 1721–1727
skin testing, 1659 Excretion film, 1720
safety evaluation, 1641–1642 biliary mixture, formulation, 1717–1718
safety issues pharmaceutical, 1656 of glucuronic acid conjugates, 323 parameters, 1716
categories, 1656 of glutathione conjugates, 323 rheological curves, 1713
Good Manufacturing Practices (GMP) urinary, of sulfate conjugates, 323 application of, 1713
for, 1656 Exogenous compounds, 310 Bagley equation, 1714
International Conference on deleterious effects of, 310 Herschel–Buckley model, 1713
Harmonization (ICH) guidance, 1656 effect of, physicochemical properties, 310 power-law model, 1713
IPEC-Americas Significant Change Exogenous pyrogens, 3054 rheological properties
Guide for, 1656 Exopeptidases, 2717 measurement of, 1715–1716
quality, 1656 Expert Working Group on Drug Quality systems, practical characterization of,
safety of, 1615–1616 (EWG-Q), 1962 1716–1717
screening, 3933 Explosion-proof devices, 2879 theory and characterization of,
selection, 1618–1619, 3642, 3933 ExplotabÕ, 1292, 3554, 3562 1712–1714
criteria, 1638–1640 disintegrating power, 3231 twin-screw, stability of, 746
solid dosage forms, 4049 rough surface, 3231 Extrusion–spheronization pelletization
sourcing, 1619 starch derivatives, 3231 axial extruders, 2656
in spray-dried powders Exponentially Weighted Moving Average classification, 2656
formulation parameters, 1650 (EWMA) control chart, 3503 critical factors in, 2660
oxidation, 1650 Extemporaneous liquid preparations, 2644 excipients, microcrystalline cellulose, 2659
particle size, 1651 Extended least squares (ELS) method, formulation components of, 2659
properties of, 1650 2756 gravity-fed extruders, 2656
spray-drying process, 1650 External stimuli triggered systems, 1328 counter-rotating cylinders, 2657
surface denaturation, 1650 Extra low carbon (ELC) grades, 790 counter-rotating gear cylinders, 2657
stability, 1962, 1614 Extracellular adhesion glycoprotein, 1328 rotary cylinder, 2657
stabilizing effects of, 1652 Extracellular calcium concentration, 2672 rotary gear extruders, 2657
stabilizing freeze-dried therapeutic proteins, Extracellular cellular fluid volume, 2634 in pharmaceutical industry, 2657
1648 Extractables marumerizer=spheronizer, 2657
stablization strategies, 1612 definition, 1693 friction plate, 2659
starch, 415 glass containers, 1710 residence time, 2659
sterilization by autoclaving, 1613 potential leachables, 1693 multistep process, 2656
supplier, 1638–1640 toxicological evaluation of, 1697 physicochemical properties, 2659
surface morphology, 43 Extractables=leachables processing equipment, 2656
surface properties of, 35 analysis of, 1698 radial-type extruders, 2658
surface structure of, 37 analytical techniques for, 1708 ram extruders, 2656
surface texture of, 1648 gas chromatographic analysis of, 1699 formulation development, 2657
in tablets and capsules identification and quantitation of, 1697 rheological properties, 2657
capsule-filling process, 1646 inorganic, 1709 screw-fed extruders
tableting types and functions, 1647 trace elements, 1710 shape transitions, 2658
for tablet production, 1646 liquid chromatographic analysis of, 1703 sustained-release characteristics, 2659
thermal analysis techniques for methods for, 1709 twin-screw extruders, 2658
pharmaceuticals, 3742 qualitative=quantitative, 1698 Extrusion–spheronization process, flow chart
tonicity adjusters, 1627, 1629 regulatory implications of, 1696 of, 747
toxicity testing, 1658 thermal analysis techniques, 1708 ExuberaÕ, 2705
toxicological issues with, 2777 thermogravimetric analysis (TGA), 1708 Eye drop
transdermal formulations, 3362–3365 with plastic components, 1694 difficulties in, 1923
types of changes Extracts in elderly patients, 1922
equipment, 1657 pilular, 957 solutions
packaging, 1657 powder, 957 birth and rise of, 1220
process, 1657 semiliquid, 957 in sterile dosage form, 1220
scale, 1657 Extractum. See Extracts. suspension dosage forms of, 1220
site, 1657 Extrapyramidal symptoms, 1042 Eye irritation
specifications, 1657 Extruders, 1712 in rabbits, 1425
types of, 1633, 1646 Nica system, 1723 testing, 1659
cosolvents, 422, 1623 selection of, 1726 Eye medication, delivery of, 1220
hydrophilic surfactants, 422 types Eye piece reticule, 2583
lipophilic surfactant, 422 ram, 1726 Eye syndromes, dry, 1220
validation, 1962 rotary-cylinder, 1724
Excipient-free tissue plasminogen activator, rotary-gear, 1724–1726
aggregation of, 1818 screen, 1722–1724 Fabric absorbents, in wound dressing, 1026
Excipient-related toxicity, 2779 for pharmaceuticals, 1722–1727 Fabrication process, 1470

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I56 Index

Factorial designs FDA issues in advertising=promotion of drug Feedback-regulated drug delivery systems,
active factor determination from, products 1099
2456 accelerated approval drugs, 62 Feed hopper designs, in roller compactors,
Box–Wilson design, 2458–2459 company-issued press releases, 61 3164
center points in, 2457 comparative claims, 61 Feed screw design
data analysis of, 3002 direct to consumer promotion, 62 horizontal twin auger, 3171
Doehlert design, 2459–2460 internet, 62 of roller compactor, 3167
hybrid designs, 2459 off-label promotion, 60 Feed water, quality of, 2880
quantitative process studies, 2455–2456 launch materials, 60 Felbinac, solubility of, 2930
statistical experimental designs, 2458 pharmacoeconomic and quality of life Femoral bifurcation, 1159
two-level fractional, 2456 claims, 61 Fenofibrate, 3348
effects from, 2457 pre-approval promotion, 61 Fenoprofen calcium, 1125, 2007
two-level full, 2455 reminder advertisements, 61 Fenoprofen salts, hydrate forms of, stability
Failure Mode and Effects Analysis (FMEA), scientific exchange, 62 of, 3184
2252 video news releases, 61 Fenoterol, 2109
Fallback delivery system, for asthmatics, FDA Modernization Act (FDAMA), 60, Fentanyl
1284 2620, 2629 anesthesia and analgesia, 2129
Falling ball viscometers, 3142 FDA regulation clinical study, 2129
newtonian materials by, viscosity of, of advertising and promotion of drugs, 59 concentrations, 2130
3142 for biological products, 59 delivery from IDDS, 2129–2130
Falling film evaporator, suitable for viscous of drug marketing practices, 64 hydrochloride solution, 2130
liquids, 3881 in internet, 65 in vitro and in vivo flux values of,
Famotidine, continous infusion of, for drug sample delivery, 64 2130
1287 for medical devices, 59 Fentanyl citrate, 2125
Fan heaterfuzzy rules to control, 2404 for pharmaceutical promotion, 59 Fentanyl hydrochloride, 2125
Faradaic electrochemical cell, 1518 in drug labeling, 59 Fentanyl OraletÕ, (Abbott labs), 999
Faradaic electrochemical techniques, of prescription drug, 59 Fenton reaction, 142
1517–1519 principles of, 59–60 Feret’s diameter (df), definition of, 639
Far-infrared region, 3405 for veterinary drugs, 59 Fermentable dietary fibre, 2872
advantage of, 1517 F-distribution, 3492 Fermentation, 259–260
Faradaicto-charging-current ratio, 1495 critical values of, 3493 Fermi levels
Faraday cup detector, 1974 two variance, evaluation of, 3492 valence electron energy state, 1535
Faraday effect, 449 variance ratio statistic, 3492 versus vacuum energy level, 1535
Faraday’s law, 1518 Febrile reactions, 3052 work function, 1535
Farmacopea de los Estados Unidos Fecal material Fermi resonance, 3407
Mexicanos (FEUM), 2856 absence of, 1301 Ferning, 1341
Fast-Flo lactose, 3277 rectal cavity of, 1302 Ferranti–Shirley cone and-plate viscometer,
Fat and gastric homogeneity, 2868 Federal Administrative Procedures Act, 1881
Fat-soluble medications, 1905 1781 Ferrite, applications, 792
Fatty acid dimers, or (FAD), in Federal agencies, 1779 Ferritic stainless steels
polyanhydride preparation, 184 department of agriculture, 1779 application, 789
Fatty acid esters, biodegradable, 1314 Environmental Protection Agency, characteristics, 789
Fatty acid methyl ester (FAME) technique, 1779 Ferro-GradumetÕ, 991
932 Federal Clean Water Act, 2882 Fertility hormones, 269
Fatty acids, 1624, 3747 Federal Food, Drug and Cosmetic Act, Fetal organogenesis, 1415
biosynthesis of, 230 1787 Fette EU1, D-tooling, 441, 3784
and derivatives in pharmaceutical quality standards, 1787 Fever
formulations, 975 excipients, 1787 induction
flow rates of, 3278 therapeutic products, 1787 LPS-binding protein (LPB), 3053
oxidation of, 1615 Federal Food Drug and Cosmetic Act, mechanism of, 3052, 3053
in tableting applications, 975 1110 proinflammatory cytokines
in vegetable oils, 978 Federal Food, Drug, and Cosmetic (FD&C) pathogenesis of, 3054
Fatty alcohols, 975 Act, 649, 1779, 2419 receptor cells, 3053
in ointments or creams, 975 drug, 1782 reticuloendothelial system, 3053
Fatty amines, 975 Durham–Humphrey amendments, 1780 Fiber filters, viscous oils, 3888
oils, waxesand neutral lipids, 975 Harrison–Kefauver amendments, 1780 Fiber optic assembly, application of, 3437
Fax transmissions, 555 Federal Regulation (CFR) code of, 707 Fiber optic fluorescence spectrometers,
FDA. See Food and Drug Administration. Federal Trade Commission (FTC), 58, 3401
FDA approved labeling, of drug products, 2424 Fiber optic fluorometer, 3401
59–60 soaps regulatory control for, 800 Fiber optic probes, 3377
FDA’s Center for Drug Research and Federation of Crude Drugs Association of Fiber optic probe, spasmoctyl samples by,
Evaluation (CDER) Japan, 2837 analysis of, 3384
pharmaceutical products by, regulation of, Feedback concentration control, Fiber optic sensors, 3401
59 implementation of, 867 applications, 3402

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I57

Fiber optic technology, for in situ UV assay, [Film-coating] [Filtration]

916 equipment, 1729–1732 sterilization of liquids by, 3886
Fibroblast growth factor (FGF), 1153 film formers, 1732 theories, 3886
Fibroblast growth factor-sucralfate, 1221 materials, 1732–1735 unit process in pharmacy, 3886
Fibrocystic breast disease, 72 of oral solid dosage forms, 1729 Financial analysis model, 3017
Fibrous absorbents physical aging, 1743 Financial analysis tools, 3017
gauze and cellulose wadding, 1027 plasticizers in, 665, 1733 Fine particle dose (FPD), 2090
gauze and cotton tissue, 1027 effectiveness of, 1736 Fine particle fraction (FPF), 1429
in wound dressing, 1025–1026, 1027 problems encountered during, 1742–1743 Fine particle mass (FPM), 2278
Fibrous polymers characteristics of, 1742–1743 Fine particle sedimentation, monitor of, 2587
alginate dressings, 1031 defects, 1742 Fining process
in wound dressing, 1031 solvents, 1732–1733 agents, 2511
Fick’s laws, 1082, 1438, 2339 systems, 665 Fire-resistant construction, 2882
of diffusion, 212, 1492 lakes and inorganic pigments, 666 Firewall, equivalent security guard, 2556
Fickian diffusion, 2666 viscoelastic properties of, 3144 First law of thermodynamics, conservation of
and drug release, 181 Film–tablet adhesion, 1739, 1741 energy, 393
kinetics, 2669 Filter First-order electrode, definition of, 1503
mechanism, 2032 air, 3888 First-order phase transitions, features of, 2935
Field-flow fractionation devices, 2590 borosilicate materials, 1749 First-principle design, implementation, 865
Filamentous fungal keratitis, in horses, 3973 classification, 3887 First-principles models, application of, 858
Filamentous glycoproteins, 2715 gravity filters, 3887 Fischer projection, 2144–2145
File room model and security, 2552 pressure filters, 3887 Fish, drug disposition in, 3943
File security, 2556 vacuum filters, 3887 Fisher 344, 1416
Filled containers, 379 configurations, 1748 Fisher subsieve sizer, 2592
Filled drug product containers, identification depth, 1748 Fiso Neb nebulizer, 2105
of, 1945 contaminants, 1748 FisonairÕ spacer, 1542
Filler=binders cylindrical filter elements, 1752 FITC-horse ferritin, 1356
classes of substances, 3228 depth retention, 1748 FITC-horseradish peroxidase, 1356
compression stages, tablets, 3654 dirt-load capacity, 1748 Fitting core-shell models, nanoparticles
coprocessed materials, 3229 disadvantages of, 1748 established by, 1066
dilution potential, 3228 fiber matrix, 1748 Fitzpatrick Co. roller compactor, 3174
magnifications, 229 fibrous materials, 1749 Fixed wavelength detectors, 534
of pharmaceutical technology, 3228 filtration method, 1754, 3886 Flame atomic absorption spectrophotometry,
requirements, 3228 lenticular modules, 1752 3368
tablets in compression of, 3228 glassfibre materials, 1749 copper in, operation screen for, 3369
Filling integrity testing materials, 1749 Flame ionization detection (FID), 1699, 3799
capsule, 408 media, 3887 Flame ionization detector, 466
machines, 409 fixed, 3887 in gas chromatography, 197
mandrels constituents of, 378 flexible, 3887 Flame photometric detector (FPD), 466
powder, 409 rigid, 3887 Flame photometry
Film membrane, 1748 advantages for, 1762
aqueous polymeric dispersions, 1730, preparation, 3887 analytical technique for, 1759
1776 probe methods, 4 applications of, 1760
disadvantage of, 3414 small-scale, 3887 atomic absorption, 1759
dropwise condensation, 3873 sterilizing grade filters, 1754 bulk drug monographs, 1761
heat transfer from, 3873 sterilizing grade membrane, 1748 formulation monographs, 1761
of mixed vapor, 3874 types of, 1748 high temperature, 1759
of pure vapor, 3874 adsorptive, 1749 low-temperature, 1759
dry and hydrated, mechanical properties of, constituents, 1749 pharmaceutical analysis, 1760
1738 narrow size distribution, 1749 quantitative tool, 1759
evaporators, 3880 validation for reagents, 1761
formation for polymers, 3413 in sterilizing grade filters, 1752 spectral properties, 1759
polymeric, mechanical properties of, pharmaceutical processes, 1752 with spectrophotometric technique, 1759
1736–1738 Filtration, 383, 3386 working principle, 1759
for resins, 3413 air mechanism of, 3887 Flame photometer, 1759
for strong materials, 3881 centrifugal, 3888 automated models of, 1760
types, 3881 methods, 3886 instrumentation of, 1759
climbing film evaporator, 3881 cake, 3886 Flammable solvents, quarantine storage of,
falling film evaporator, 3881 clarification, 3886 2879
rising-falling film evaporator, 3881 depth, 3886 Flammable waste products, 2879
Film–capsule adhesion, 1737 microorganism removal, 1748 FlashdoseÕ (Fuisz Technology), 999
Film-coated drug products, 1735 membrane types of, 3028 FlashtabÕ (Prographarm), 999
Film-coating resistance of crystal, 861 Flashtab technology, tablets manufactured
colorants in, 665 separative process of, 1748 with, 1110

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I58 Index

Flat bottom flowmeter, 3276 [Flocculated suspensions] Fluid-bed technique, 2330

Flat filter membranes stability, by brookfield viscometer, Fluid corporate nervous system, 2558
microbial detection, 1748 3608 Fluid dynamics, analysis of, 4045
specifications, 1748 viscosity of, 3608 Fluid energy grinding, 3600
Flavin-dependent dehydrogenase (FAD), Flocculating agents, of suspensions, 3606 for drug particle size reduction, 3600
115, 321 Flocculation Fluid extracts
Flavonoids, 147–148, 2911 capillary, 3141 formulations, 957
antioxidants, 143 continuous phase, 1555 liquid preparations, 957
and activity of enzymes, 143 dilatant, 3132 Fluid flow
biosynthesis of, 234, 235 Newtonian, 3130 analysis, 3868
cinnamoyl structure, 143 Non-Newtonian, 3130 Bernoulli’s theorem, 3862–3863
dietary intake, 143 pseudoplastic, 3130 in bodies, 3867
quercetin, sources of, 143 reversible process, 1555 flow energy, 3862
and terpenoids, benefits of, 72 slit, 3141 frictional losses at pipe fittings, 3866
Flavor and Extract Manufacturers FlocorTM, 1643 kinetic energy evaluation, 3862
Association (FEMA), 801 Flossing, 898 laminar flow, 3863, 3865
Flavor–drug interaction, 2232 Flow degradation energy in, 3865
Flavor modifiers, 1763 avalanching, 2354 flow changes, 3865
Flavoring agents cascading, 2355 measurement, 3863–3865
in liquid dosage forms, 2225 cataracting, 2355 venturi meter, 3863, 3864
of pharmaceuticals, 2225 centrifuging, 2355 mechanical energy, 3862
Flavors, 1763 measurements, 1512 in pipes, 3865
agents, 1763 parameters, 3278 pressure drop calculation, 3866
artificial, 1765 rolling, 2354–2355 in pipe frication, 3866
classification, 1764 slipping, 2353–2354 pressure energy, 3862
natural, 1764–1765 sustain release agent, 2081 pumps, 3869
characteristics, 1766 Flow cytometry, 3402 impeller pumps, 3869
components, 1763 Flow device, with movable orifices, 3281 positive-displacement pumps, 3869
feeling factors, 1763 Flow enhancers See Glidants. resistance of, 2590
odor, 1763 Flow-injection analysis (FIA), 1499 Reynolds number, 3865
taste, 1763 immunoassays with, 2050 streamline flow
definition, 1763 of reagents, 2050 mathematical analysis, 3865
drug products, 1772 technique of, 1512 Poiseuille’s law, 3865
enhancers, 1770 Flow injection renewable surface in tube, 3865
carboxylic acids, 1770 immunoassay (FIRSI) velocity, 3865
citric acid, 1770 with fluorescence detection, 1577 volumetric flow rate, 3866
food, 1770 Flow-through cells, for liquid sample analysis, tubes in noncircular cross section, 3866
pharmaceutical industries, 1770 3413 turbulent flow, 3863, 3865
sugar, 1770 Flow types, static flow, 3280 unit mass volume, 3862
foods aroma of, 1763 Flowmeter, dimensions of, 3281 unit operation in pharmacy, 3862
in industry, 1763 Floxin otic drops, 2481 volumetric flow rate, 3863
ingredients, 1766 Fluid air micronizers, energy source in, 2880 Fluid flow dynamics, 2640
instrumentation methods, 1763 Fluid and solid boundary, heat transfer, 3871 Fluid thioglycollate medium (FTM),
modification technology, 1770 split filter design in, 1445 2287
chemical modification, 1771 Fluid-bed coating application, 766 Fluid vesicle dispersions, 1120
complexation, 1771 Fluid-bed coating types Fluid extractum. Fluid extracts.
microencapsulation and coated systems, bottom spray methods, 2319 Fluid glycerates, 957
1771 tangential-spray coating method, 2319 Fluid glyceratum. See Fluid glycerates.
solubility-limiting methods, 1770–1771 top-spray methods, 2319 Fluidization
vesicles and liposomes, 1771 Fluid bed dryers, 1444–1446, 3892 technology, 1773
pharmaceutical, 1764 coolants in, 1444 types of, 1773
liquids, 1764 drying tablet granules, 3657 Fluidization velocity, for spherical particles,
pastes, 1764 Fluid bed-drying, 1444 1445
preparation, 1767 for organic vapor applications, 1445 Flunisolide, 2110
solids, 1764 of spherical particle, 1436 Flunixin meglumine
potentiators, 1770 Fluid-bed granulation, 2456 by intravenous injection, 3959
primary taste, 1766 tablets manufacturing process, 3657 in cows, 3959
selection of criteria for, 1767 Fluid-bed granulator–dryers, 2876 treatment of mastitis, 3959
taste, 1763 Fluid bed granulators, 744, 2875 Fluorapatite, definition of, 891
taste-masking effects, 1770 Heinen continuous, 745 Fluorescein isothiocyanate (FITC), 203
FlavorTech, ODT technologies in Fluid bed processes, 1773 adiabatic process, 3197
development, 1111 particulate design and preparation, airflow design parameters, 3198
Flocculated suspensions, 3602 1776–1778 air velocity design parameters, 3198
preparation, 3603 Fluid bed reactor, 1134 air velocity versus bowl capacity, 3198

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I59

[Fluorescein isothiocyanate (FITC)] [Fluorescence detectors] [Fluorinated elastomers]

bovine albumin, 1356 high-performance liquid chromatography constituents of, 2242
dextran, 3579 (HPLC), 3402 Fluorinated surfactants, 344
energy transfer, 3197 Fluorescence immunoassays (FIA), 2054 Fluorocarbon–hydrocarbon diblock
equipment design criteria, 3198 fluorescent labels, 2054 compounds, 344
equivalent drying efficiency, 3198 fluorogenic molecules, 2054 Fluorocarbon—perfluorooctyl bromide,
fluidization air velocity, 3198 heterogeneous, 2054 342–345
fluidized material, 3197 homogeneous, 2054 Fluorocarbons, 1617
heat convection, 3197 immunochemical binding system, 2054 emulsions, 349
heat transfer rate, 3198 photoexcited fluorescence measurement, liquids, 1430
heating design parameters, 3198 2054 Fluoroelastomers, saturated elastomers,
heating versus bowl capacity, 3198 spectral characteristics of, 2054 1467
mass transfer rate, 3197 Fluorescence polarization immunoassay, Fluorogenic detection reagents, 542
moisture content 204 Fluorogestone acetate, 1351
near-infrared spectroscopy (NIR), 3198 Brownian motion of, 2055 Fluorophore, 1972
periodic measurement of, 3198 fluorescence immunoassays, 204 decay characteristics,, 3403
rate-limiting diffusion, 3197 fluorescence transition moment, 2055 isothiocyanates of, 203
scale-up of, 3198 phenomenon of, 204 luminescent label, 203
thermodynamic treatment, 3197 photoselected molecules, 2055 Fluoropolymers, melt-processible, 2241
Fluorescence, 3387 physical principle, 2055 Fluoroquinolone, antibiotic, 1190, 2807
absorption of ultraviolet, 201 polarized emission, 204 Fluoroquinolone otic preparations,
advantages of, 201 Fluorescence spectra 2481
applications, 1973 chemical structural aspects of, 3389 Fluoroquinolones, 1399
biomedical trace analysis, 199 and high-performance liquid and metals, 1397
cells, 3398 chromatography (HPLC), 3402 antiinfective agent, 1397
characteristics of, 3388 hydrogen-bonding solvents in, 3390 constituents of, 1397
chemiluminescence, 204 of pharmaceutical compounds, 3389 Fluororelastomers, 2237
comparison, 1974–1975 pH effects in, 3390 Fluosol, assessment of, 342
as chromatographic detection device, 452 solvent effects in, 3390 Fluosol-DAÕ, 1550
decay time, 1973, 3388 temperature effects in, 3392 FluosolÕ, 1643
detector, 201 Fluorescent analytical methods, in Flurbiprofen, 1078
efficiencies fluorescence emission pharmaceutical sciences, 3387 salts melting point for, 3180
CD(FDCD), 452 Fluorescent antibody tests, 1144 Foam aerosols, 997
excitation wavelengths, 201 Fluorescent cell-sorting, 3402 Focal necrosis, 1142
filter instruments, 201 Fluorescent in situ hybridization (FISH), 932 Focused beam reflectance measurement
flow cells, 201 Fluorescent microscopes, in biomedical (FBRM), 4082
fluorimeters, 201 sciences, 3400 definition of, 860
fluorimetric detector, 201 Fluorescent molecules method, 4082
halogenated solvents, 201 acid-base reactions of, 3391 Foil gages, application, 3685
headspace oxygen analyzer based on, concentration of, 3391 Foil preparation, 2978
1973 lifetimes of, 3391 Folate receptor, 1328
high-performance liquid chromatography, Fluorescent optical sensors (FLOPS) difference levels of, 1328
201 in analytical spectroscopy, 3402 mediated endocytosis, 1334
intensity of, 3392 instrumentation, 3401 Folate-targeted liposomes, deposition of,
intensity quenching, 1972 fiber optic, 3401 1333
light emission, 204 lasers, 3401 Folic acid antagonist, 1134
limitations, 1973 optical filters, 3401 Follicle stimulating hormone (FSH), 70, 1429,
measurements, 1065 Fluorescent quenching technologies, 3049 2748
measurements, 201 Fluorescent probes, 2054 Fomentations
non-halogen-containing solvents for, Fluoride mouthrinse, 895 constituents, 957
201 products effect of, 895 narcotic drugs, 957
of 2-naphthoic acid, 3389 solutions, 895 Food, 1779
operation technique, 1971–1972 Fluoride tablets, application of, 893 and acarbose (Precose), 1397
pH dependence of, 3391 Fluorimeters, 201 treatment of diabetes mellitus, 1397
polarization, 1576 analysis in aqueous solutions and Alendronate (Fosamax), 1397
quantitation for, 201 excitation wavelength, 201 bioavailability of, 1397
quantum yield of, 3389 fluorescence detector, 201 and miglitol (Glyset), 1397
quenching, 542, 3391 operation of, 201 treatment of diabetes mellitus, 1397
quenching sensors, 1973 pulsed-source, 2056 antiinfective agents, 1397
schematic diagram of, 201 radioactive decay curves, 2056 colorants, 649
sensors, 3401 sensitive and selective drug absorption, 1397
wavelengths of, 3389 steady state, 2056 excipients for, 1638
Fluorescence detectors, 304, 534 Fluorinated elastomers, 2242 in gastrointestinal (GI) tract, 1397
of aromatic molecules, 3389 carbon–fluorine bonds, 2242 interaction study, assessment of, 2922

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I60 Index

[Food] [Food and Drug Administration (FDA)] Fotus. See Fomentations.

in vitro models, 2819 in United States, 1891 Fourier-transform ion cyclotron resonance
biorelevant dissolution models, 2819 in vitro release rate, 1320–1321 (FTICR), 3806
gastric fluid simulation, 2819 controlled-release dosage forms, 3190 applications of, 3806, 3807
on absorption, influence of, 2824 enteric-coated dosage forms, 3190 Fourier analysis techniques, 2583
mixture, viscous rheology of, 1720 immediate-release dosage forms, 3190 Fourier transform (FT) NMR spectroscopy,
on pharmacokinetics, impact of, 2819 solid oral dosage forms, 3190 3297
poorly water-soluble, effect of, 2819 in vitro dissolution, 3190 Fourier transform-near-infrared
Food additive petition, drug product, 1658 in vitro release-rate testing, 3190 spectroscopy, 3631 (FT-NIR)
Food Additives Amendment of 432, 1958 in vivo bioequivalence, 3190 Fourier transform infrared microscopy
Food and color additives regulation, 649 Food and drug administration modernization for bulk drugs analysis, 3417
Food and Drug Administration (FDA), 1, Act, 61, 1780, 2841, 3191 non-invasive method, 3417
591, 649, 1104, 1572, 1622, 1650, biologic products, 1781 for structural identification, 3417
1657, 1931, 1941, 1955, 2384, 2468, FDA’s policies, 1781 Fourier transform infrared (FTIR)
2629 FDA’s regulatory activities, 1780 spectrometer, 3405, 3407, 3410
abbreviated new drug applications Food and Drug Cosmetic Act (FD&C), chromatographic instruments, for
(ANDAs), 1782, 1783 1929 structural characterization, 3417
activities, 1779 Food and drug law, 2841 advantages of, 3410
applications of, 3190 Foods, drugs, and cosmetics (FD&C) applications of, 3405
blend uniformity draft guidance, colarants, 651 chromatographic technique, 3405
2968 allergic-type reactions by, 650 double-beam, 3410–3411
CDER, 1783 aluminum lakes, 656 in industrial on-line process control, 3410
chemical testing, 3190 fading characteristics of, 669 optical diagram of, 3411
clinical investigators, 1926 physical properties for, 669 single-beam, 3410
devices, 1780 blue, 648 Fourier transform infrared spectroscopy
drugs, 1780 dyes, 651 (FTIR), 305, 701, 1648, 1708, 1830,
federal agencies, 1779 fading characteristics of, 669 1839
foods, 1780 orange, 648 measurements, 1648
clinical supply inspections of product label of, 650 protein–excipient hydrogen bonding, 1648
manufacturers, 592 solubilities of, 667 protein secondary structures, 1648
for commercialization, 2625 stability data, 667 Fourier transform spectrometers, 3375
cystic fibrosis treatment of, 1650 yellow, 652 Fourier’s equationof heat conduction, 1435
in Dietary Supplement Health and Foods, drugs and cosmetic act (FDCA), Four sector mass spectrometer=Fourier
Education Act (DSHEA), 1789 432, 2904 transform mass spectrometer,
diffusion analyses, 3190 Food-effect studies 2264
diffusion testing, 3190 design of, 2821 analog detection device, 2265
drug formulations, 2626 timing of, 2821 biological matrices, 2264
drug packaging guidelines and regulations, Foods, drugs and cosmetics (FD&C) yellow, constant neutral loss scanning capabilities,
2526 652 2267
drug regulation, 1782 Footand-mouth disease virus, 3911, in mice, mass accuracy, 2264
exclusivity, 2619 3912 radio flow detector, 2264
Federal Food, Drug & Cosmetic Act for cystic fibrosis, 273 selective=sensitive detector, 2267
(FFDCA), 1781 for flavor modification technology, 1771 ultrahigh resolution, 2264
fine particle fraction, 1650 Forced convection heating process, 3513 Fox–Flory equation, 399
guidance Forced-circulation evaporators, 3880 Free energy perturbations (FEP), 724
industry, 1781–1782 Forced-convection batch sterilizer, 3513 advantages of, 724
reviewers, 1782 Force-feeders, in roller compactors, 3165 equilibrium constants, 724
guidelines, 1404 Force transducer, as strain gauge, 3666 mutations process for, 725
for carcinogenicity studies, 689 Forcolonic bacterial metabolism, 2871 Fractal
for OTC medication labels, 2418 Fordermal therapy, 996 applications in pharmaceutical and
international harmonization, 1788–1789 Foreign compounds, 310 biological, 1796
innovative programming, 2166 deleterious effects of, 310 aerosol delivery, 1799–1800
manufacturing regulation, 3076 excretion of, 310 atomization, 1799
NDAs, 1783 glucuronidation, 318 dissolution, 1796–1798
new drug application (NDA) submitted to, Foreign molecules, degradation of, 23 granules, 1798
561 Form factor morphological change, skin, 1803–1804
in Nutritional Labeling and Education Act analytic function of, 1055 protein, 1800–1802
(NLEA), 1789 definition of, 1053 tabletting, 1799
oral solid dosage forms, 1893 Formazan, 151 aerosols, 1800
organization classification of, 1 Fosinopril sodium, 2942 analysis, 1791
physical tests, 3190 Fosphenytoin boundary and surface, 1791
Public Health Service Act, 1781 constituents, 2251 atomic force microscopy (AFM), 1794
and stability information systems, 733 prodrug, 2251 gas adsorption, 1793
stability long-term, 3 therapeutically active compound, 2250 image analysis, 1791–1792

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I61

[Fractal] Freeze–drying process Frusemide, 2862

mercury intrusion porosimetry, 1793 for preparation of FTC regulation, 63–64
charcoal, 3277 ketoprofen and dicumarol dispersions, in advertising and promotion of
cromolyn sodium, 3277 775 drugs, 64
dimension, 1791 Freeze–thaw in prescription drug advertising, 64
geometry, 3277 cycles, 1824 FTC versus FDA regulation, 63–64
lactose, 3277 procedures, 1117 in advertising and promotion of
Fractal dimensions and aerosol stability, 1826 drugs, 64
characterization, 1799–1800 Freeze fracture technique, 1119 Full-stream trough sampler, 2964
Fraction cosolvent, 811 Freeze-fracture electron microscopy, Fumaric acid (FA), in polyanhydride
Fractionating powders, 2589 1319 preparation, 184
Fractionation process, 2586 use of, 1058 Functional proteomics, 3041
Fracture mechanisms, 2972, dendrimer Freezing, 1808–1812 Functions of, 143
fractured, 884 concentration, 1808–1809 Fungi-nailÕ, 2423
Fragility, definition of, 86 crystallization of excipients and Funnel flow device, with interchangeable
Fragrances, microencapsulated, 2330 consequences, 1809–1810 orifices, 3281
FrandolÕ, 1086 instability, 1810–1811 Furanocoumarins, biosynthesis of, 235
isosorbide dinitrate-releasing TDD system, parameters, 1811 Furosemide, 2862
1086 pH during, shifts in, 1810 dissolution activation energies for, 778
Free energy of phosphate buffer solutions, 1810 polymorphs relaxation times of, 2942
of adsorption, 1172 stabilization during, 1825–1826 Fusion process, in ointment preparation, 3266
diagram, 87 Freezing-point depression Fusogenic lipids, 644
Free energy–temperature relationship, 86 colligative property, 3772 Fuzzification, 2404
Free-fall blenders cryoscopic constant, 3772 Fuzzy logic, 2403
bin blenders, 3205 crystallization, 3771 applications, 2403–2404
double-cone blenders, 3205 definition, 3772 tablet disintegration time by, 2403
V-blenders, 3205 molal depression constant, 3772 tablet hardness desirability by, 2404
Free-fall tumbling mixer non-volatile solute, 3772 Fuzzy rules, for fan heater control, 2404
chaotic conditions in, 2967 osmometers, 3776 Fuzzy sets, 2403–2404
Free plasma abciximab concentrations, 2810 Raoult’s law, 3772
Free radical, 3542 solid and liquid phases, 3771 GAB equation, 2372
in carcinogenesis, 139 thermodynamic principles, 3772 Galactomannam, cross-linked, 1237
definition of, 141 Freezing-point osmometry, 3776 Galactosidase protein, 121, 1414
detection and characterization of, 151 French paradox, 2439 Galenical Developmental System (GSH),
diseases associated with, 148 Frenkel–Halsey–Hill (FHH) theory, fractal 1668
sources of, 141 geometry, 1793 Galenicals, 948
in stroke, 139 Frenkel defect, 3545 extraction, 948
toxic biological response, 139 Freon propellants, 2093 pharmaceutical preparation, 948
Freeze concentration in amorphous system, Frequency distribution curves, 2974 Gall bladder contraction, 2872
1809 Frequency modulation spectroscopy (FMS) Gallopamil, 2125
Freeze-dried products, 2979 applications, 1971 Galvani potential difference, 1502
formulation and stability, 1820–1830 comparison, 1974–1975 Galvanic corrosion, of metals in seawater, 783
relationships between formulation and in non-destructive gas concentrations Gamma-aminobutyric acid (GABA), 76, 1043
process, 1821–1822 monitoring, 1971 release of, 76
Freeze-drying, 1834 operation principle, 1971 Gamma camera imaging, 3095
advantage, 1107, 4113 Fresnel reflection, 3377, 3379 Gamma cyclodextrins, 682, 687
development and scale-up pathway, Fresnel’s equation, 448 Gamma-hydroxybutyrolactone and
1834–1836 Fretting corrosion, 783 hydroxybutyric acid, 186
equipment influences, 1846–1847 Freund’s adjuvant, 1418 in polyorthoester degradation, 186
excipient, 1822 Fricke dosimeter, 3543 Gamma interferon, 1614, application of,
formulation design and product Front–end multiple quadrupole ion source, 2743
characterization, 1836–1837 3047 Gamma-interferon, application of, 2743
parameters, processing, 1838 FrontlineÕ, 3981 Gamma-linoleic acid (GLA), 72
plants, 1807 FrostaTM, ODT technologies in development, Gamma rays
process, 1808–1820 1112 in nuclear medicine, 3085
process, lysozyme model, 1838 Froude number Gamma-sorbitol, 3681
process development considerations, compaction force, 3194 as tablet diluent, 3681
1842–1845 granulation, 3194 crystalline forms, 3681
product design, 1836 range of, 4087 spray-dried varieties, 3681
product preparation, 1837 in scale-up technology, 3195 Gamma scintigraphic imaging, study of, 2868
solid, 3893 versus bowl capacity, 3195 Gamma scintillation counting, 1150
tablet matrix principle, 1107 Frozen ethanol layer, 2318 Gamma-rays and X-rays energies of
X-ray powder diffractometry (XRD) Frozen maltodextrin systems, 1811 measurement of, 3086
technique, 4113 Fructose, isomerization to, 1614 Gamma-scintigraphy, assessment with, 1290

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I62 Index

Ganciclovir, 1224 [Gas chromatography (GC)] Gas-permeable membrane, 1508

Gang test assays, 3510 of organic volatile impurities, 501–514 Gas-sensing configuration, 1508
Garbling process, 2905 pharmaceutcial raw materials and dosage Gas-sensing electrodes, 1508
Gargarisma. See Gargles. forms, 518 Gas-tight locking syringe, 1970
Garlic (Allium sativum), 72 pharmaceutical, 500–518 in drug delivery systems, 3571
Gas adsorption chromatography (GSC), 463 volatile matter, intermediates, 487–496 in paracetamol precipitation, 3572
Gas adsorption studies, procedures for, 2590 portable, 1970 for solute precipitation, 3570
Gas antisolvent recrystallization (GAS), quantitative and qualitative analysis, 474 capillary, 3571
3570 separation process, 196 coaxial, 3571
Gas chromatography (GC), 196 separation techniques, 473–474 ultrasonic atomizers, 3571
advantage of, 463 solvents for, 196 Gastrointestinal tract (GI), 426
alcohol content stationary phase absorbed in, 1242, 1298
determination of, 499–500 system components=equipment, 466 absorption window of, 2866
water in pharmaceutical raw materials automatic injection, 468–469 anatomical arrangement, 3943
and dosage forms, 497 columns, 469–473 monogastric species, 3943
aqueous solutions, 196 on-column injection, 468 ruminant species, 3943
argon ionization detector, 198 detectors, 469, 470–471 antigens in, degradation of, 3918
sensitivity, 198 flow control, 467 blood perfusion of, 1288
analyzers, 1970 gases, 466–467 capacity
applications for inlet splitter, function, 468 in adult cattle, 3947
chromatographic and radiochemical oven and temperature controls, 469 in goats, 3947
purity, 483–484 sample inlets, 467 in sheep, 3947
autosampler, 1970 split injection, 468 disease, 2664
for biomedical applications, 197 splitless injection, 468 microflora in, 2631
capillary column, 473 temperature programming, 473 occluding junctional complex (OJC),
advantage, 464, 472, 516 thermal conductivity detector, 197 1230
development, 463 sensitivity, 197 drug absorption, 1396, 2631
high resolution capability of, 516 theory, 464–466 epithelium, 24, 25
rate theory for, 466 types of detectors used in, 196 hypersensitivity reactions, 47
thickness, 472 volatile by chemical derivatization, 196 medication in, 1905
components, 1968 Gas chromatography=Mass spectrometry mucosa, 23
determinations, types, 514 (GC=MS) mucosal immune system, 3917
drawback, 1969 abietic acid, 1699 pharmacokinetic interactions, 1396
efficiency of, 466 amphetamines in hair, determination of, pH of, 22
electron capture detector 3571 physiology of, 22
carrier gas flow rate, 198 applications, 1699, 1704 secretions, 24
halogen atoms, 198 atmospheric pressure chemical ionization, structure and motility, 23
polar functional groups, 198 1705 therapeutic activity, 1396
sensitivity, 198 negative ion mass spectra, 1704, 1705 Gas phase polymerizations, 3543
flame ionization detector, 197 positive ion mass spectra, 1704 Gas pycnometer, 3283
sensitivity, 197 chromatogram of, 1694 Gas streams, spherical particles in,
gas flow rates, 467 cold filtration technique, 1702 1437
headspace oxygen determination, 1968 elastomeric packaging component, 1699 Gastric absorption, of acidic drugs, 23
headspace analysis, 474, 500 of extractables=leachables, 1699 Gastric acid secretion
hydrogen flame detector, 198 ionization processes cephalic phase, 24
sensitivity, 198 chemical ionization, 1699 gastric phase, 24
injection port of, 199 electron ionization, 1699 intestinal phase, 24
injection process, 468 molecular ion confirmation, 1701 phases of, 23
isothermal, 473 from pharmaceutical packaging materials, Gastric acidity, 2820
for lipid analysis, 981 1699 nocturnal patterns of, 2872
and mass spectrometer (MS), 198 pyrolysis, 1708 Gastric carcinoma, 150
analysis of trifluoperazine, 198 qualitative analysis, 1699, 1704 Gastric emptying
application of, 198 thermal desorption, 1708 delayed, 1253
assay method for, 198 for drug analysis, 3572 fat on, effect of, 2868
chemical ionization, 198 Gas–liquid chromatography (GLC), 463, posture and gravity on
ionic mass-to-charge ratios, 198 2849 effect of, 2868
in pharmacological research, 198 flash evaporation, 196 phases of, 2871
radio-labeled trifluoperazine, 198 liquid stationary phase, 196 process of, 2867
time-of-flight, 198 non-volatile substances, 196 variability in, 2825
mobile phase transports in, 1968 boiling point, 197 Gastric emptying
organic compounds, 197 liquid column coating, 197 nature of, 1246
pharmaceutical identification of raw melting point, 197 process rate, 1423
materials and dosage forms thermostable, 197 time (GET), 1246, 2631, 2820
determination of, 485–486 separation process, 196 Gastric irritation, 605, 1245

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I63

Gastric juice, 24 [Gelatin] Gel foam (UpJohn), 1883

constituents of, 24 formulations, 1869–1871, 1872 Gel-forming solutions, 1220
ionic composition of, 24 exposure of, 1861 Gelled petrolatum base, 3259
volume of, 2872 in hydrogel preparation, 190 Gelling agents, 3264
Gastric motility, disease affect, 2871 physiochemical properties of, 421 polysaccharide, 1885
Gastric mucosa polypeptide backbone, 1863, 1865 Gelling materials, 1289
epithelium of, 2713 primary amino groups of, 1869 Gel permeation chromatography (GPC),
irritant to, 1253 sponge absorbable, 1883 532
Gastric proteases, oral drug administration types, 1863 of polymers
Gastric residence time, 1291 animal-derived excipients, 1642 for separation, 532
Gastric retention, 1253 capsules 992 polar organic mobile phases in, 532
Gastric ulcer healing, model for, 296 dissolution characteristics of soft, 1867 pharmaceutical, 1886
Gastroduodenal ulcers, 1242 hard, 992 carbomer, 1887
Gastroduodenostomy, 218 soft, 992 hydroxypropyl cellulose, 1886
Gastroesophageal reflux disorder, 1242, 2871 Gelatin–acacia complex coacervation, 2330 hydroxypropyl methylcellulose, 1886
Gastrointestinal absorption, 428, 940, 3945 Gelatin capsules, hard, moisture in, 3383 structurers, 532
from gastrointestinal tract, 3945 Gelatin-gum arabic system, 2316 in two-phase systems, 3264
Gastrointestinal bleeding, 944 capsules, 406, 1616, 2085 Gel-phase nuclear magnetic resonance, 3456
Gastrointestinal disturbance, 3003 bioavailability of digoxin, 3948 Gel-state mucus layers, 2680
Gastrointestinal degradation, 2674 dimensions of, 408 GelucireÕ, 1294, 1258
Gastrointestinal disease, effect of, 2871 drying process, 408 in chloroform, 4069
Gastrointestinal disorders, patients with, film, 407 Gelucire 44=14, 778
1304 and hypromellose, 407–408 Gene
Gastrointestinal effects, 73 colorants, 407 biochemical analysis of, 1901
Gastrointestinal fluid, 426, 1093, 1294 pigments, 407 biological utilization of, 1901
Gastrointestinal irritation, absence of, 1294 preservatives, 407 complex process, 1898
Gastrointestinal motility, 214, 1017 polyethylene glycol (PEG) in, 407 delivery vehicles, 1065
changes in, 2872 solutions, 407 difference between antisense oligo and,
process of, 1244 storage conditions, 413 936
Gastrointestinal physiology, 2713 Gelatin membrane filter (GMF), 2310 and DNA delivery, to skin, 2751
Gastrointestinal stromal tumors, types of, Gelatin nanospheres drugs, 1899
1326 for gene delivery, 1186 encoding, 1134
Gastrointestinal symptoms, 368 sealing process, 422 epigenetics controlled by, 1898
Gastrointestinal therapeutic systems (GITS), shell ingredients, 424 field of, 2752
992 Gelatina. See Gelatins, 957. for cell immortality, 264
Gastrointestinal transit, 1246 Gelatum. See Gels. Gene–gun administered plasmids, 1355
effect of, 2871 Gel-diffusion test, 1418 goal of, 2751
variation in, 2866 method, 3060 isolation, 260
Gastrointestinal ulcerative lesions, 681 Gel-formers large-scale analysis is, 1901
Gastroretentive systems absorbable, 161 method of, 645
application of, 1253 application of, 161 mutation-like effects, 1897
design of, 1253 burn wound healing, 162 mutations, 438
Gaucher’s disease, 2473 hemostatic application, 162 physiological effects, 2796
Gaulin-type homogenizers, 1997 insulin controlled release systems, 162 protein function, 1901
Gaussian distribution, 1898 misoprostol protein identification of, 1901
Gaussian functional link network, 2402 release of, 163 structure of, 1054
Gaussian-shaped waveform, 1495 periodontal application, 161 therapy, 263, 929
Gauss-Newton approximation, 2763 as sealants for microporous implants, transcription controlled by, 1898
Gavage dosing, 1423Gear pump, 1731 163 variability of, 1897
Geiger–Mueller counters See GM counters. skin wound repair, 162 Gene-deleted vaccines, 3910
Gelatin, 991, 1718, 1861, 1883 tissue adhesive formulation, 162 Salmonella typhimurium, 3911
characteristics Gel film ophthalmic (Upjohn), 1883 pseudorabies virus, 3911
film forming properties, 992 Gel filtration chromatography (GFC), 532 thymidine kinase gene, 3911
non-toxic, 992 calibration methods Gene expression microarrays (GEMs), 2198
reversible phase change, 992 broad molecular weight, 532 Gene products, indirect, fermentation as,
soluble in biological fluids, 992 primary, 532 2832
cross-linking secondary, 532 Gene-specific oligonucleotides, 2796
rate of, 1866 universal, 532 molecular mechanisms, 2796
reaction, 1864, 1865 for separation, of water soluble biopolymer quantitative real-time PCRs
digestion, by enzymes, 1861 Gel-forming hydrophilic polymers, 1881 amplification of, 2797
drug delivery system, 1644 Gel-forming polymers, 1875 enzyme reverse transcriptase (RT), 2797
factors inducing, 1865–1867 Gel-forming substances gene expression kinetics of, 2797
film absorbable, 1883 concentrations of, 1880 for single nucleotide polymorphisms,
forms of, 992 pharmaceutical uses, 1881–1889 2797

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I64 Index

[Gene-specific oligonucleotides] Genome 264, 1420 Gibbs–Helmholtz free energy, 401

gene activity regulation of, 1898 definition, 930 Gibbs phase rule
LUX primers, 2797 protein complement of, 3041 for salol–thymol system, 762
Scorpion primers, 2797 techniques, 1897 thermal analysis techniques for drug,
self-quenching primers, 2797 Genotoxic compounds, 436 3733
tissue-specific DNA modification, 1898 Genotoxic contaminants, 442 Ginger (Zingiber officinale), 72
X-chromosome inactivation, 1898 Genotoxic estrogen metabolites, 2443 Ginger jake paralysis. See Jamaica ginger
analysis and interpretation of, 2797 Genotoxicity paralysis.
biochemical mechanisms, 2796 assays, 437 Gingival crevicular fluid, 904
complementary DNAs (cDNAs), 2796 battery, 438 Gingivitis-reducing properties, 898
drug resistance development of, 2797 potential, 2774 Gingko biloba, 72–73
General Agreement on Tarrifs and Trade testing, 569 Ginseng, 73
(GATT), 3718 Genotyping definition of, 73
Generally recognized as safe and effective analysis of PCR products, 2796 GISSI-prevenzione trial, 2439
(GRASE) drugs, 2420 DNA sequencing, 2796 GITSÕ, 1246
Generally Regarded As Safe (GRAS), 2273 polymerase chain reaction amplification of Glactosamine, 1330
Generic antibiotic application, 3188 microsatellites, 2796 Glass
Generic Drug Enforcement Act, 1780 restriction fragment length polymorphism of amorphous multicomponent mixtures,
Generic drug products (RFLP), 2796 399
bioavailability–bioequivalency, 1892 techniques for, 2796 capsule device, 1214
measurement, 1893 Gentamicin, 162, 1020, 1353 classification of, 1276
business strategies of, 1894 drug delivery ophthalmic, 1222 containers, and polymer processing,
development, 1891 elimination glomerular filtration, 3965 179
regulatory process, 1891 half-life of, 3963 in metered dose inhalers (MDIs),
side-effects profile as, 1891 in nebulizers, 2096 2274
testing of, 1892 nebulization flow rate, 2097 DSC scan of, 400
therapeutic profile as, 1891 volume distribution, 3964 electrodes, 1507
Generic manufacturer, 1891 Gentamicin soaked hydrogel lenses, 1221 formation, for small-volume parenterals,
Generic pharmaceutical industry sssociation Gentamicin sulfate, plasma concentration, 1276
(GPIA), 2471 3955 volume changes associated with, 777
Generic pharmaceutical manufacturer, Gentian violet, 2479 high-shear mixing of polymer, 179
2616 GeodonÕ, 3361 light sensitive SVIs, 1276
for commercialization, 2625 Geographical BSE Risk (GBR), 406 melting temperature of, 776
identification of patents, 2621 GeomatrixÕ system, 991, 999, 1255 membrane, in precipitation, 1276
infringement exemptions, 2624 Geometric isomerism, 2146 surface of, 1507
innovator’s product centers, 2620 C-heteroatom double bonds, 2147 of miscible mixtures, 399
paragraph certification, 2621 in cyclic compounds, 2147–2148 of polymers
patent certification, 2621 non-mirror images, 2146 factors affecting, 180
patent infringement liability, 2624 non-superimposable images, 2146 plasticizers effect of, 180
Generic pharmaceutical products Geometric standard deviation (GSD), pH levels, 1276
application of, 4100 2093 polymer extrusion of, 179
marketing approval of, 4100 Geriatric dosage form, 998–999, 1905, quality control of, 1276
testing of, 4102 1922–1923 solutions and glass transition temperature,
Genetic algorithms Geriatric dosing, 1905 776
and neural networks, 2402 Geriatric pathology, 433, 434 solutions and suspensions, 776–777
optimization, 2403 Geriatric patients solutions formation, 777
Genetic algorithm similarity program dosage forms of, 1922 solutions vs. solid solutions, 777
(GASP), 4018 eyedrops, 1923 definition of, 776
Genetic diseases inhalers, 1923 transition temperature (Tg), definition of,
genetic variability effects, 1900 Germ cell mutagenicity, 1413 86
medical diagnosis, 1899 Germ line mutations, 1413 transition temperature and polyorthoester
mendelian disorders, 1899 Germaben II-E, 3270 composition, 186
phenotypic expression, 1899 Germall 115, 3270 transition temperature, 399, 2079, 2925
Genetic engineering, 260 Germall II, 3270 Type I, II and III, 1276
methods, 355 German pharmacopeia, 2212 transitions, 1122
Genetic material, cloning of, 260–261 Gerteis Co. roller compactor Glassy–rubbery transition, 2032
Genetic optimization for ligand docking compaction, 3174 Glatt Multicell production line, 745
(GOLD) program, 4027 diagnostic feature functions, 3173 Glatt Powrex systems, 3194
Genetic polymorphism, 586 feeding system, 3174 Glaucoma
Genetic skin disease, 3821 granulating units, 3174 treatment of, 944
Genetic therapies, 278 Ghoose–Crippen atom types, 4015 delivery, 1223
Gengrafcyclosporin A formulation, 3349 Giant papillary conjunctivitis (GPC), 2204 Glaucoma drug. See Pilocarpine.
Genistein, 2441 Gibbs free energy, 636, 2219 Glaucoma filtration surgery, 1223
in cancer prevention, 2443–2444 derivative of, 2935 GLC (Gas-liquid chromatography), 2

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I65

GliadelÕ Glucuronyltransferase, 2635 Glycoprotein

Glidant action, mechanism of, 3280 Glutamate hydrolase, conjugate of, 1146 cell adhesion molecules are, 1141
Glidants, 988, 990, 3279 Glutamic acid, 42 mucus a biochemical component of,
colloidal silicone dioxide, 3280 and naltrexone, 177 1346
magnesium stearate, 3280 Glutaraldehyde, 2025, 2316, 2332–2333 structure, 1347
starch, 3280 protein crosslinking by, 2317 surface of, 2720
tablets assessing method 3659 sporicidal activity of, 2988 Glycosaminoglycan, 1884, 2028
colloidal silicain 3659 Glutathione chondroitin, 2431
talc, 3280 biliary excretion of, molecular-weight, 323 glucosamine, 2431
Glioblastoma multiforme 179 conjugation reactions, 318 matrix formation, 2436
recurrent, 186 conjugation reactions of detoxication, Glycosides, 2911
and GliadelÕ therapy, 179 319 Glycosidic ether oxygens, 682
Glioma cells, receptors on, 1328 in asymmetric centers, 318 Glycosidic prodrugs
Global and National Commerce Act, in stereochemistry, 319 synthesis of, 1256
elctronic signature in, 2560 in stereospecificity, 319 Glycuronoalycan, 1883
Global planning, effective, 570 epixodes, 318 Glycyrrhiza, powders of, 2971
Globular proteins non-specific enzymes, 318 Glydant, 3270
cell membrane, 25 in oxidation, epoxides, 313 Glyoxal bis(dially acetal) (GLY), 2023, 2025,
intrinsic proteins, 25 phase I biotransformation reactions, 318 2026
Glomerular filtration, 2636, 3095 substitution reactions, 318 structure, 2024
Glomerular filtration rate (GFR), 576, 586, Glutathione reductase, 1136 Glysters. See Enemas.
1020 Glutathione S-transferases Goblet cells, 1177
cats, 3962 cell structures, 319 Gold colloid, preparation, 637
cattle, 3962 endoplasmic reticulum, 319 Gonadotropin-releasing hormones (GnRH),
dogs, 3962 isoenzymes, 319 1355
goats, 3962 Glutathione synthetase, 1019 Gonadotropin-releasing hormone (GnRH),
humans, 3962 Gluteal administration, technique for, 2645 1099
measure of, 3032 Gluten protein, malabsorption of, 1615 for prostate carcinoma, 1099
pigs, 3962 Glutoid capsules. See Capsules, glutoid, 958. Good automated manufacturing practices
sheep, 3962 Glycation, definition of, 286 (GAMP) forum, 1
Glomerular function, 2636 Glycerides derivatives, 3747 electronic records and signatures in, 6
Glove piston effect, 2139 Glycerin, 3360 environment, 2
Glucopyranose, 672 sweeteners, 1770 in Europe, 2
Glucosamine, 73, 2431 Glyceritum. See Glycerites. Good clinical practice (GCP), 2167
active constituents in, 2446 Glycerogelatins, 958 guidelines, 1925
applications, 2436 Glycerogelatinum. See Glycerogelatins. non-compliance, 1929
and chondroitin combination, 2446 Glycerol, 2382 obligations, 1925
clinical trials, 2436 Glycerol monostearate, 1734 investigator, 1925
combination therapy, 2437 melting and rheological properties of, 767 monitor, 1925
disaccharide of, 3054 Glycerol palmito stearate, 2978 sponsor, 1925
effects, 2436 Glyceryl methacrylate (GM), 2028 regulations, 1925
GAG matrix formation by, 2436 Glyceryl monocaprylate=caprate, 14 Good Clinical Practice (GCP), Code of
incidence, 2435–2436 Glyceryl monooleate, 1294, 1071, 1075–1076 Federal Regulations (CFR) part 11, 4
Glucosamine=Chondroitin, 73 Glyceryl trinitrate, 991, 1071 Good clinical practice (GCP) regulations,
Glucosamine=Chondroitin Arthritis in dosage forms, 3949 3064, 2486, 1783
Intervention Trial (GAIT), 2436 in metered-dose aerosol preparations, Good information practices
Glucosamine sulfate, 2436 1076 guidelines for, 4101
Glucose pharmax tablets of, 1076 use of, 4101
cyclic, 1617 sublingual administration of, 1076 Good laboratory practice (GLP), 434, 2775
polymer, 3060 tablets of, 1076 development on, 3064
subcutaneous injection of, 116 Glyceryl trinitrate tablets, monograph for, regulations, 2486
Glucose-containing formulations, 2381 3653 Good Laboratory Practice (GLP), Code of
Glucose-6- phosphate dehydrogenase Glycine, 1820 Federal Regulations (CFR) part 11, 4
(G6PD), 1019 carbonyl resonance of, 3301 application of
GlucoWatchÕ G2TM Biographer, 3849 domestic animal species, 3963 computer systems, 1936
Glucuronidation, 2635 steric hindrance and polymer degradation compliance with, 1931
conjugation reactions, 317 effect on, 188 levels of, 1933
of endogenous substrates, 318 sucrose system, 1821 maintenance of, 1938
foreign compounds, 318 Glycine–citric acid requirements of, 1932
of tertiary amines, 318 Glycocalyx, 23, 2715 environment, 1936
Glucuronides intramolecular rearrangement, Glycogen synthesis, 317 equipments for
318 Glycolate buffer, use of, 389 calibration and validation of, 1936
Glucuronidic prodrugs, 1255 Glycolipid Compounds, 976 centrifuges and balances, 1936
Glucuronosyl transferase, 2444 Glycolmethacrylates, 2028 computer maintenance, 1936

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I66 Index

[Good Laboratory Practice (GLP)] Gradient elution, 526 [Granulators]

high-pressure liquid chromatography Grains, 2353 kinetic, 3194
(HPLC), 1936 convection in, 2356 parameters for
golden rule in, 1935 dispersion in, 2356 principle of, 3194
independent quality assurance unit (QAU), mixing, 2352 process forces in, 3193
1931 scaling, 2359 machine design parameters, 3195
management-designated archivist, 1938 segregation, 2358 mechanical, 744
monitoring authority, 1932, 1933 Gram-negative bacteria, 2791, 3052, 3900 shear forces, 3194
non-clinical safety, 1932 cell membrane of, 3053 tip speed
objective of, 1931 in nebulization equipment and patients, definition, 3194
pitfalls and benefits of, 1938 3856 versus bowl capacity, 3194
principles of, 1931, 1932 Gram-negative microorganisms, elimination Granule bond formation
stability testing, 1937 of, 4043 bonding, 3161
test substance, 1937 Gram-negative vibrio bacteria, 3057 characteristics, 3161
Good manufacturing practice (GMP), 29, Gram-positive bacteria, 3900 deformation, 3161
223, 306, 737, 1006, 1783, 2167, 2286, Grans method fragmentation, 3161
2850, 2875, 3064 Nernst equation, 3761 rearrangement process, 3161
controls, for clinical supplies, 591 potentiometric titration, 3760–3761 Granule growth mechanisms, 36
guidelines for, 34 Granular bed, 2353 Granule preparation, roller compaction in,
buildings and facilities, 594 Granular behaviors, analysis, 2353 2409
change control, 596 Granular devices, 2352 Granule properties
complaints and adverse reactions, 598 Granular effervescent salts, 958 disintegration time, 2406
control of incoming materials, 594 Granular flow, 2352 friability, 2406
data integrity considerations, 598 Granular material, phases in, 2352 hardness, 2406
deviations and failure investigations, Granular properties, empirical measurements Granules, 95, 1798
597–598 of, 3281 compression properties of, 38
equipment, 594 Granular solids in drug, 4068
integrity of, 598 flow properties of, 1798 drug release dependent on, 4068
laboratory controls, 596–597 suspension properties of, 1798 glycerol monostearate in, 4068
master (and batch) production and Granular state, definition, 2352 properties of, 767
control records, 593–594 Granulate, particle size of, 4078 stearic acid in, 4068
production and process controls, 595 Granulated dry syrup wax
qualification and validation activities, agglomerated, 3233 congealing method, 4068
594–595 constituents, 3233 fusion method, 4068
quality organization, 593 granules, 3233 melt granulation technique, 4068
sterility assurance, 595 particle size, 3233 Gran’s plot, for potentiometric titration,
technology transfer plans and reports, Granulating agent, in plasticity of powder 3761
598 mass, 3670 Grapefruit juice, 1397
to clinical supplies, applicability of, 591 Granulation bioavailability of, 1397
equipment cleaning processes of, 1580 adsorption in, 36 calcium channel blocking agents for, 1397
drugs, 1580 of alpha-lactose monohydrate, 746 components of, 1397
pharmaceuticals, 3928 colored wet, 666 serum concentrations, 1397
regulations for medical devices, 3928 continuous, 745 Grapeseed (Vitis vinifera), 73
microbiological, 2290 disintegrants in Graphite furnace technique, 3369
of sterile drug products, 2290 dry, 745, 3159 autosampler, 3369
activities, 5 effervescent production of, 746 disadvantage of, 3369
electronic records for, 5 equipment, classification of, 744 Zeeman background correction, 3369
European guidance, 2 high-shear mixer, 767 GRAS, in cosmetic products, 1317
system, 4 incorporation of, 3562–3564 Gratings
Good manufacturing practices guide liquid, 745 diffraction, 3397
pharmaceutical excipients, 1638 pharmaceutical methods of, 3159 excitation, 3397
Good manufacturing practices (GMP), powder, 3160 factors in, 3397
pharmaceutical, 380 properties, 3160 reflection, 3396
Good Manufacturing Practice (GMP)-quality tablets manufacturing process, 3657 transmission, 3396
drug, 2487 tablets manufacturing process, 988–989, Gravity-flow auger sampler, 2964
Good post-marketing surveillance practices 3657 system, 1010
(GPMSP), 1980 taste masking drug, 1106 Gravity filters, 3887
Gordon–Taylor equation, 399 wet, 3159 Gravity settle plates, 2307
Goserelin acetate, delivery of Granulators, fluid bed, 744 Greek mythology, 2901, 2999
ZoladexÕ for, 178 Froude number, 3194 Green tea (Camellia sinensis), 73
Gouy–Chapman layer Heinen continuous, 745 causes, 2438
electric double layer high-shear, 3193 consumption, 2438
pharmaceutical particles, 4117 dynamic, 3194 fermentation, 2438
Gowning and degowning procedures, 2878 geometric, 3194 flavonoids of, 2438

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I67

[Green tea (Camellia sinensis)] HaloLiteTM Hb-encapsulation efficiency, 356

manufacture, 2438 components, 2111 Hb solution, preparation of, 355, 357
Grinding pharmaceutical materials performance comparison, 2116 Headspace oxygen
equipment Halofantrine analysis, 1967
ball mills, 3896 antimalarial drug, 1610 determination, 1968
colloid mills, 3896 Halogenated compounds and halides, 1633 measurements, 1967
edge- and end-runner mills, 3896 Haloperidol (Haldol), 1047 sampling tools comparison points, 1974
fluid energy mills, 3896 from haloperidol decanoate, 311 accuracy, 1975
hammer mills, 3896 Haloperidol decanoate, 3362 linearity, 1975
pin mills, 896 ester hydrolysis of, 311 quantitation limit, 1975
roller mills, 3896 Hamaker summation method, 640 techniques
vibratory mills, 3896 Hancock and Zografi study electrochemical methods, 1970
Griseofulvin, 231 drug substances, amorphous state of, 3736 fluorescence quenching methods,
absorption of, 28, 2870 glass transition changes, 3737 1971–1974
biosynthesis of, 231 Hanging mercury drop electrode (HMDE), frequency modulation spectroscopy
drug solubility, 28 1492 (FMS), 1971
hard gelatin capsule formulation for, 1671 Hanks’ Balance Salt Solution (HBSS), headspace gas chromatography (GC),
micronization of, 774 3999 1968
particle size, 28 Hard candy lozenges, 2231 quadrapole mass spectrometric analyzers,
solid dispersions, 41 in-process testing, 2234 1974
in solid solutions, 28 raw materials, 2231 Health and drug regulatory authorities, 4101
Gross vesicle morphologies, examine of, 1058 stability, 2235 Health and Human Services (HHS), 64
Group-deuterated version, of surfactant, 1051 Hard gelatin capsules (HGCs) Health care delivery systems, 1977
Growth factor receptors, MoAbs with coloring economic system, 1977
interaction of, 1137 colored imprinting inks, 666 geography, 1977
Growth factors (GFs), 697 FD&C or D&C colorants, 666 infrastructure, 1977
Growth harmone, rectal absorption of, 1301 titanium dioxide, 666 political system, 1977
Growth hormone, human watersoluble dyes, 666 traditions and conventions, 1977
in immunosupressed rats, 179 liquid filling, advantages of, 4070 Health care financing
GSH conjugation catalysis, in acetylation, Hard-particle methods, 2355–2356 administration of, 1989
319 characteristics, 992 children’s health insurance program, 1990
Guanine nucleotide regulatory protein gelucire bases within, 4070 cost-effectiveness, 1991
activation of, 3108 liquid filled, 992 factors for, 1989
Guar gum, 1235 surfactants in, 3592 health care revenues, 1989
polygalactomannans in, 1236 and tablets medicare program, 1990
Guggenheim and deBoer equation, 4051 lubricants in, 3592 Organization for Economic Cooperation
for vapor adsorption, 4051 preparation, 3592 and Development (OECD), 1989
Guided evolutionary simulated annealing thixotropic formulations into, 4070 prospective payment system (PPS), 1991
(GESA) algorithm, 2402 with waxes, 4070 in United States, 1989
Gum rubber, 1468 Zanasi, 4070 Health Care Financing Administration
Gums, types of, 958 Hard tablets, by amorphous lactose, 3742 (HCFA), 52, 1987
Gurana flavor, 1770 Harmonic alternating current, 1498 Health care products, 57
Gut-associated lymphoid tissue, 2714 Harmonization, 927 Health care services, 1977
Guttae. See Drops. characteristics, 1958 economic and social survival for, 1987
concordant methods, 1963 financing, 1977
drug quality standards, 1958–1960 death, causes of, 1992
Haber–Weiss reactions, 142, 2862 for drugs, 1955 morbidity in, 1992
HabitrolÕ excipients, 1955, 1963–1964 mortality in, 1992
nicotine controlled dose of transdermal, factors, 1956 services-utilization requirements, 1991
1086 goal of, 1960, 2855 Ministry of Health and Welfare (MHW),
Haemophilus influenzae, 2479 history, 1957 1980
vaccine, control of, 4101 international, 1788 organization, 1977, 1987
Hagen–Poiseuille’s law, 1713 international conference on, 3071 structure, 1977
Hair colorants, 800 interpharmacopeial, 1961 technological and scientific developments,
Hair follicles pharmacopeial, 1955 1988
human skin, 3968 process, 1638 in United States, 1977, 1987, 1991
Halide ion-selective electrodes, 1512 prospective, 1958 utilization of, 1987
Halitosis, 900 retrospective, 1958 Health care system
Hallucinogens, 1201 scale for, 1960 administering medications, 55
acute effects of (bad trips), 1046 values, 1955 in adverse drug reactions (ADR), 55
chronic effects and reactions, 1047–1048 Hatch–Waxman Amendments, 2619 in bar coding, 55
classification, 1045 process patents, 2626 medication errors, 55
history and nature of, 1045–1046 Hatch–Waxman provisions, 2624 for monitoring and reporting adverse
plant and mineral, 1045 Hazardous waste, recycling of, 2882 drug events, 55

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I68 Index

[Health care system] Heat of fusion rule, 2936 Hemoglobin

pharmacy-based intravenous medication, Heat of transition rule, 2936 aggregation of, 1824
55 Heat-resistant glasses, 2514 infusion, 370
unit dose medication distribution, 55 Heat-shock proteins, 145 Hemolysis
American cultural, 1985 Heats of vaporization, of solvents, 1440 adverse pathological conditions, 3768
American geography, 1986 Heat stability, depyrogenating heat-resistant cosolvent induced, 815
American lifestyle, 1986 materials, 3055 dynamic flow conditions, 815
American topography, 1986 Heat transfer, 1435–1438, 3869–3875 measurements, 815
Clinton administration plan, 1993 to boiling liquids, 3872 phenomenon, 1272
Egalitarian principles, 1985 in evaporator, 3879 physiological significance, 3768
government and political policy, 1985 with forced circulation, 3873 of red blood cells, 815
industrialization of, 1986 temperature, 3879 static flow conditions, 815
Libertarian principles, 1985 in vertical tubes, 3873 Hemolytic anemia, 1019
medicare program, 1986 crystal growth by, 3885 Hemolytic transfusion reaction, 335
national and state governments in, 1986 fictitious film, 3871 Hemophilus type b conjugate vaccines, 4101
political and economic policy film coefficient, 3871 Hemorrhagic shock patients, 360
developments, 1985 from film condensation, 3873 Hemorrhoidal vein, 1300
social development, 1985 fluid to solid boundary, 3871 absorption by, 1306
of United States, 1987 fluid velocity, 3872 Hemorrhoids, treatment of, 1300
Utilitarianism, 1985 dropwise, 3873 Hemostatic application, gel-formers, 162
Health insurance mixed vapor, 3874 Henderson–Hasselbach equation, 213, 387,
complexity and concentration of, 1988 mechanisms, 3869 1074, 1508
Independent Practice Association (IPA), conduction, 3869 for acid and base, 1074
1988 convection, 3869 Henry’s law
integrated health system, 1988 radiation, 3869 colligative property, 3770
Prefered Provider Organization (PPO), nucleate boiling, 3872 definition, 3770
1988 in pipes, 3870 interionic interactions, 3770
Health maintenance organizations(HMOs), by radiation, 3874 non-volatile solute, 3770
67, 1998 black body, 3874 thermodynamic activity, 3770
group model, 1988 calculation, 3875 Hepaplus liposom, 1128
out-of-pocket payments, 1990 exchange of, 3874 Heparin-coated poly(methylmethacrylate)
staff model, 1988 Stefan–Boltzmann law of, 3874 nanoparticles, by emulsion
types of, 1988 steady nondirectional, 3870 polymerization, 1188
Health Products and Food Branch (HPFB), through composite walls, 3870 Hepatic asialoglycoprotein receptors, 1330
in Canada, new drug application through plane walls, 3869 Hepatic blood clearance, 575
(NDA) submitted to, 561 in tubes, 3870 Hepatic cirrhosis, 580
Health Protection Branch (HPB), 1572 turbulent mixing, 3871 Hepatic clearance, 3031
Health registration regulations, 2604 unit operation in pharmacy, 3869, 3879 Hepatic cytochromes, 577
Health system models in vertical tubes, 3873 Hepatic extraction ratio (ERH), 577
mixed systems, 1977 Hectorite, 1889 Hepatic first-pass effect, 2678, 2741
private sector, 1977 Hectorol, 3347 partial avoidance of, 1303
state health insurance program, 1977 Height equivalent of theoretical plate Hepatic first-pass extraction, drugs with,
state ownership and control, 1977 (HETP), 464–465 1305
Health Technical Memorandum (HTM), Heinen continuous fluid bed granulator, 745 Hepatic first-pass metabolism, 2664, 2672
2291 Hele-Shaw configurations, 2367 oral administration, 25
Heart arrhythmia, 3436 Helicobacter pylori, 3918 Hepatic flexure, 2871
Heart transplant rejection infection, 2873 Hepatic-gastrointestinal (GI) metabolism,
antimyosin Fab, 1156 HeliosTM gun system, 1320 1339
immunosuppressive treatments for, Helmholtz–Smoluchowski equation, 4117 Hepatic haemodynamics
1156 electrophoretic mobility measurement, 4119 assessment of, 2824
Heat and mass transfer coefficients, air zeta potential determination, 4119 Hepatic metabolism, 580, 1610, 2635
velocity, 3884 Hematological analyses, 362 ciprofloxacin, 1399
Heat capacity Hematology, evaluation of, 686 definition of, 1247
of pharmaceutical materials, by DSC, Hematopoiesis–erythropoiesis, 363 enoxacin (Penetrex), 1399
399 HEMA–VP–styrene, 2028 process of, 1247
thermodynamic analysis, 89 Heme oxygenases, 142 serum concentrations of medications,
Heat energy in circuit, flow of, 1438 Heme synthesis, microbiological, 358 1399
Heat flow signal, Fourier transformation of, Hemihedral quartz crystals, 2142 Hepatic monooxygenases, 2672
395 Hemihydrate, sweetener aspartame as, 4112 Hepatic plasma flow, 3031
Heating Heminevrin capsules, 3347 Hepatic portal system, 1610
components, 2175–2176 Hemodilution therapy, 363 Hepatic portal vein, 216, 1244, 1610
ventilation and air conditioning (HVAC) Hemodynamic effects, 371 concentration, 584
system, 8 Hemodynamic measurements, systemic, 366 Hepatic presystemic metabolism, 1303
Heat-labile enterotoxin (LT-B), 3912 Hemodynamics, normalization of, 362 Hepatic tissue, of drug reservoir, 1191

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I69

Hepatic tumor-feeding artery infusion, Heterogeneous amperometric immunoassays, [High efficiency particulate air filters]
1331 1527 filtration mechanisms
Hepatitis, 269 Heterogeneous assay, 2049 diffusion, 2174
alcoholic, 75 Heterogeneous electrochemical enzyme electrostatic attraction, 2174
chronic active, 1019 immunoassays, 1527, 1528 inertial impaction, 2174
Hepatitis A virus (HAV), 3999 Heterogeneous enzyme immunoassays interception, 2174
strains of, 4004 (HEIA) sedimentation, 2174
Hepatitis B, 1527 classification, 2052 leak-integrity testing, 2304
vaccine, 274 competitive assays, 2052 performance certification, 2174–2175
requirements for, 4101 non-ompetitive assays, 2052 in pharmaceutical applications, 2174
Hepatitis B surface antigen-encoding plasmid, Heterogeneous fluorescence immunoassay, shipment, storage and handling, 2174
1355 2057 velocity and uniformity testing, 2304
Hepatitis B surface antigen (HBsAg), 3998 Heterogeneous immunoassay High-extraction-ratio drug, 588
Hepatitis B virus (HBV), 3911, 3912, 3998 fluorescence immunoassays, 204 High frequency oscillating devices, 2112
Hepatitis C virus (HCV), 3998 fluorescent labels, 203 AeroDoseTM, 2112
Hepatocyte radioimmunoassay, 203 metered solution inhaler (MSI), 2112–2113
membranes, 75 Heterogeneous liposome immunoassays, vibrating membrane nebulizer, 2113
PLA matrices, incorporation in, 190 2060 High frequency sonophoresis, 3835
rat, 1419 Heterogeneous nucleation processes, mechanisms, 3838
transplanted, in new liver-like tissue 841–842 High-impedance voltmeter, 1502
formation, 190 Heterogeneous subpopulation of cells, High mass resolution=high mass accuracy
Hepatotoxicity, 74, 76 1145 tandem mass spectrometer, 2264
screening for, 2196 Heterogenous non-competitive immunoassay, in metabolite characterization, 2264
Heptafluoropropane, 2107 1567 quadruple time of flight (Q-Tof )
Heptapeptide Heterologous polyclonal antibody, instrument, 2264
cyclic, oxidative degradation of, 1614 1570 High-methoxy (HM) pectin gels, 1884
Herbal(s) HETeronuclear CORrelation (HETCOR), High-performance liquid chromatography
medications, 51 3447 (HPLC), 198, 463, 526, 538, 700,
medicines, 66 Heteronuclear multiple bond coherence 1499, 1589
abuse of, 2910 (HMBC), 3449 absorption photometric detectors, 199
adverse reactions with, 2908 Heteronuclear multiple quantum coherence aldoses analyses by, 456
cost of, 2910 (HMQC), 3448 apparatus constituents of, 199
manufacturers of, 2906 Heteronuclear single quantum coherence assays, 916
propagation of, 2909 (HSQC), 3448 autosamplers, 533
risks and realities of, 2906–2910 Hevea Brasiliensis (rubber tree), 1479 Beer’s law for, 200
products, 2902 Hexachlorophene (pHisohexÕ), 2478, 2633 biomedical trace analysis, 199
as dietary supplements, 2904 bacteriostatic activity, 2478 p-bonding electrons, 199
significance, 66–68 for surgical prophylaxis, 2478 calibration in, external and internal, 528
cost, 67 Hexaketide derivatives, 231 cell path length, 200
demographics of users, 67 Hexamethylene tetramine (HMT), 2997 columns, 528
prevalence of use, 67 Hexyl-insulin monoconjugate 2 (HIM2), composition of, 1703
supplements 2707 computers in, 535
efficacy of, 69 HicapsealTM, 412 detectors, 533–535, 1703
manufacturers, 69 Hierarchical non-linear mixed-effects characteristics of, 533
Herbal anxiolytic kava, 2910 modeling, 2806 types, 199
Herbal drugs, 2902, 2906 High-affinity binding reagents diffusion rate of, 199
interactions, 2908–2909 chemical stability, 1576 fixed-wavelength filter photometer, 200
Herbal fen-phen, 74 thermal stability, 1576 fixed-wavelength UV detectors, 200
Herbalism, 2901 High-angle annular dark field (HAADF) advantages of, 200
Herb–drug interactions, 70 detector, 2397 low-pressure mercury arc lamp for, 200
Herbs High-density lipoprotein (HDL) cholesterol, flow fluctuations, 200
consist of, 2908 2438 fluorescence detectors, operation of, 3402
defintion, 2903 High-density oligonucleotide arrays, 1901 for identification of drug, 3708
potentially unsafe, 76–79 High density polyethylene (HDPE), 1109, of individual components, 526
HerceptinÕ(Trastuzumab), 1630 2014, 2529, 2538 isocratic elution, 526
Heroic medicines, 1957 containers, 2542 liquid mobile phase, 1703
Heroin, 1040 High efficiency particulate air filters, 594, method validation process of, 535–536
Herpes simplex virus (HSV), 4000 748, 2140, 2881, 3513 limit of detection (LOD) in, 536
Hespan, injectable products, 1628 aerosols collection on, 2174 limit of quantitation (LOQ) in, 536
Hess’ law of summation, 393 collection efficiency, 2174 parameters in, 536
Heterobifunctional reagents, 1135, 1136 construction, 2173 mobile phase in, 526, 528, 543
Heterochiral amine, 2156 in-duct, 2182–2183 modes of, 528–529
Heterochiral ligands (HeL), 2159 ductless terminal, 2183 molar absorptivity, 200
Heterodimeric glycoproteins, 1328 filtration efficiency, 2173–2174 non-bonded electrons, 199

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I70 Index

[High-performance liquid chromatography [High shear wet granulation] Homochiral ligands (HoL), 2159
(HPLC)] raw materials for, 3197 Homogeneity, verification of, 2976
non-polar lipids analysis by, 98 reproducibility of, 3194 Homogeneous amperometric immunoassays,
optical path dimensions, 200 rheology of, 3196 1528
packing materials for, 199 scale-up effects of, 3194, 3196 Homogeneous electrochemical enzyme
in pharmaceutical analysis, 526 tableting process immunoassays, 1528
photometers types of, 200 High speed DSC. See HyperDSC. Homogeneous enzyme immunoassays
for quantitative analytical separations, High-temperature flame photometry, 1759 (HOIA), 2051
528 High-throughput organic synthesis (HTOS), antigen labeled enzyme modulator,
separation efficiency, 526–528 1365 2051
small-aperture microvolume flow cells in, application of, 1367 cloned enzyme donor immunoassay
200 bioassay network, 1366 (CEDIAR), 2052
solute molecules, 526 chemical diversity, 1367 enzyme acceptor (EA), 2052
solvent (mobile phase) delivery in, 532 chemical libraries, 1366 enzyme donor (ED), 2052
spectrophotometric detector cells, 200 combinatorial libraries, 1367 for therapeutic drugs, 2052
stationary phase in, 526, 528 compound management, 1365 enzyme labeled antigen for, 2051
system maintenance in, 537 design principles, 1367 fluorogenic enzyme substrate, 2052
techniques, for preparative chemical development of, 1366 Homogeneous fluorescence immunoassay,
separations, 528 features of, 1367 2055
thermal decomposition, 1703 molecular complexity, 1368 Homogeneous immunoassay
thermally stable compounds in, 198 organizational structure of, 1365 advantage of, 1528
ultraviolet (UV) detectors for, 200 semimicro level synthesis, 1367 excitation wavelength, 203
variable-wavelength grating fluorimeters skeletal and functional complexity, 1366 fluorescence immunoassays, 204
for, 201 structural diversity of, 1366 fluorescent labels, 203
variable-wavelength spectrophotometer, High-throughput screening (HTS)1362, 2652, fluorescent signal, 203
200 2993 Homogeneous liposome immunoassays
variable-wavelength UV detector, 201 acquisition and management costs, 1366 cytolysin mediated assays, 2060
components of, 201 advantages of, 1366 mellitin-based assays, 2060
development of, 201 application of, 4013 structural characteristics of, 2060
High performance liquid chromatography assays, 1366 Homogeneous nucleation, 838–841
coupled with tandem mass chemical diversity, 1366 Homogenization
spectrometry (HPLC-MS=MS), chemical functional groups dispersion, mechanical forces in
2263 1366 cavitation, 1996
advantages of, 2263 chemical techniques, 1366 impact, 1996
analytical technique, 2263 materials, 3452 shear, 1996
characteristic neutral losses, 2264 micro requirements of, 1366 turbulence, 1996
chromatographic conditions, 2263 molecular complexity, 1368 particle size reduction by, 1996
conjugated metabolites, 2264 techniques, 2486, 3048 in pharmaceutical suspensions, 1996
fragmentation characteristic, 2264 technlogy, 4013 rotor-stator, 1999–2000
for metabolite characterization, 2263 High velocity scanning infrared laser beam ultrasonic, 2002
software programs in, 2266 applications of, 860 Homogenizer, 3268
High performance thin layer chromatography Higuchi’s theory, 2930 applications
(HPTLC), 538 Hildebrand equation, 810 cell disruption, 1996
linear ascending development, 540 Hill equation, 573 liposomes manufacture, 1996
quantitative analysis of, 542 definition of, 1016 micro-encapsulation, 1996
sample application in, 540 Hippocampal neuronal ischemia, 367 solid–lipid nanoparticles manufacture,
with spectrometric methods, 544 Histamine H2-receptor antagonists, 2805 1996
High-pressure homogenization, 1996–1998 Histidine tablet coating dispersions, 1996
High pressure pumps oxidation of, 285 equipment considerations, 2002
for solvent delivery, in chromatography, Histidine-labeled Ni-containing agarose high-pressure, 1996
533 magnetic beads, 933 in cell disruption, 1997
High-resistance instruments, 1509 Hixson and Crowell cube root law, 909 in emulsification, 1997
High-shear granulators, 3193 HMG-CoA reductase inhibitor encapsulation efficiency by, 1998
High-shear mixer, 3195 salt form selection for, 3184–3185 Gaulin type, 1997
High shear vacuum processor, 1447 Hodgkin’s lymphoma B-cells, 1137 for homogenizing milk, 1997
High shear wet granulation Hodgkin’s lymphomas, non, 1328 homogenizing valve assembly in, 1997
equipment, 3195 Hoechst 936–937 liposomes preparation, 1997–1998
light-scattering applications, 3197 Hofmeister series, 601, 1507 microparticles preparation, 1997–1998
parameters, 3196 Hollow cathode lamp, 3367, 3395 in proteins harvest, 1998
particle consolidation, 3193 Hologram, 4017 manufacturers of, 2001
power consumption=torque, 3196 Homeopathic preparations in pharmaceutical industry, 1996
primary forces, 3193 monograph on, 2829 rotor-stator, 2000
process analytical technology (PAT), Homo sapiens, 431 Homogenous antibodies
3196 Homochiral derivatizing agent, 2156–2157 quantities of, 1132

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I71

Homologous blood transfusions, 353 [Hot-melt extruder] Human colorectal adenocarcinoma cells,
HOMO–LUMO interactions pharmaceutical processing equipment, 1335
non-traditional parameters, 717 3200 Human dermal replacement, 1035
Homopolymer scaling method for, 3201 Human epidermal growth factor, 296
definition of, 2925 twin-screw, 3200 receptor, 1137, 1328
Hookean body Hot-melt extrusion Human eye, phototopic sensitivity range of,
mechanical model for advantages, 2004 2863
steel spring as, 3134 applications, 2012 Human food-effect studies, 2817
oscillating shear stress and shear strain of, benefits biliary secretion, stimulation of, 2817
3137 anhydrous process, 2007 drug–food interaction study, 2817
Hookean elasticity, 3134 tableting problems, elimination of, gamma-scintiography, gastric investigaion
Hook’s law, 1293 2007 by, 2817
Hopper components, 3200 intergastric volume effect, 2817
and bins, design of, 3278 dosage forms by, 2004 splanchnic blood flow, 2817
for capsule filling, 3279 downstream auxiliary equipment, 2004, types of, 2817
design, 2081 2006–2007 Human food safety, 3985
to pharmaceuticals, 3279 extruder, 2004 acceptable daily intake (ADI), 3985
flow factor of, 3279 single-screw, 2005 antimicrobial resistance, characterization
flow, models of, 3278 twin-screw, 2005 of, 3987
funnel flow, 3279 extrusion screw, efficiency by, 2005 human intestinal microflora, effect on,
mass-flow, design of, 3279 in food industries, 3200 3986
segregation in, mechanisms of, 3279 limitations, 2012 maximum residue limit (MRL), 3986
stress and material strength, 3279 materials in, 2007 regression methods, 3986
for tablet presses, 3279 monitoring tools, 2004 teratogenic and carcinogenic effects, 3985
Hormone adrenaline, 249 plasticizers therapeutic antimicrobial veterinary
Hormone dependent tumors applications, 2008 medicinal products, 3987
stimulation of, 71 citrate esters, 2008 tissue residue concentrations, 3986
treatment of, buserelin in, 1078 formulation, 2008 total residue depletion curves, 3986
Hormone replacement therapy, 2869, functions of, 2008 violative residues, absence of, 3986
2441 incorporation, 2008–2009 Human genetic cell repository, 2796
Hormones, 269–270 polyethylene glycols, 2008 Human genome, mapping, 2487
fertility, 269 thermochemical stability and volatility of, Human granulate colony stimulating factor
Horny starches, 3478 2008 (g-CSF), 1613
Horseradish peroxidase (HRP), 2724 triacetin, 2008 Human growth hormone, 283, 2473, 2736
permeability coefficient of, 2725 for pharmaceutical applications, 2004 administration of, in immunosupressed
Hosokawa powder characteristic tester, pharmaceutical technology, 3199 rats, 179
3286 in plastics industry, 2004, 3200 in animal models, 2736
Hot-air laminar flow sterilization tunnels, process, 2004–2007, 3199 chemical and aggregation stability of,
3515 conditions, 2006 1828
Hot emulsions, coating with, advantages and product variability, 3200 chemical degradation of1817, 1828
disadvantages, 4072 residence time distribution (RTD), formulation, 2319
Hot-melt extruded dosage forms 3201 glycine and mannitol in, 1821
drug release rate from, 2010 screw design scale-up, 3200 subcutaneous administration of, 2736
excipients in, medicaments and functional, technique, 2004 Human hemoglobin, properties, 354
2007 technology viability, 2013 Human hybridomas, 1132
polymeric carriers in, 2009 Hot-melt fluid-bed coating process, Human immunodeficiency virus (HIV),
Hot-melt extruded drug delivery systems, drug by, 768 2473
2008 Hot melt method, 2330 types 1 and 2, 3998
Hot-melt extruded films Hot-melt microencapsulation, 2317 infected cells, 1141
adhesion of, force of, 2018 Hot stage microscopy (HSM), 700 infection, in humans and spongiform
bioadhesion testing of, 2017 Hovine flowcapsÕ, 1432 encephalopathies, 1616
Chatillon testing apparatus in, 2017 Human adenovirus types, 3909 protease
processing conditions for, 2017 Human airway, 2093 constituents of, 728
Hot-melt extruded granules, 2011 Human anti-mouse antibodies (HAMA), structure of, 728
acetaminophen release, 2019 263, 1132, 1570 symmetrical dimmer, 728
tablets preparation from, 2019 Human blood coagulation, 3057 X-ray structure of, 728
Hot-melt extruded tablets, 2008 Human calcitonin, 2748 protease inhibitor, 729, 834, 3336
Hot-melt extruded theophylline, Human cancer, causes of, 431 viral genome, 728
2011 Human carcinogen, 2927 Human insulin, 460
Hot-melt extruder Human carcinogenicity, 431 crystals, surfaces of, 845
advantages of, 3200 Human cell-based pyrogen test, 3057 Human intestinal blood flow, stimuation of,
jacketed tank equipment, 3200 Human colon carcinoma 2824
melt granulation equipment, 3200 cell line model, 2720 Human intubation method, 2720
mixing in types of, 3200 grafts, 1145 Human lymphocytes, 437

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I72 Index

Human LysPro, 460 [Humidity] [Hydrodynamic pressure-activated drug

Human monocytes, activation of, 3058 transepidermal water loss (TEWL), delivery systems]
Human nasal cavity, 2678 3823 liquid drug formulation, 1094
Human nasal cell lines, 2683 HumulinÕ N, 1266 Hydrodynamic technique, 1524
anaplastic nasal septum, 2683 Hyaluronan, 2437 Hydrodynamic voltammetry (HDV),
iopharmaceutical purposes, 2683 Hyaluronic acid, 1881, 1884 1520
transepithelial electrical resistances (TERs), drug delivery system, 1644 Hydrodynamically balanced systems (HBS),
2683 products, 274–275 988
Human nasal epithelial cells synovial production of, 2436 Hydrofibers, 1033
cellular functions, 2682 in wound management, 1035 Hydrofluoroalkanes (HFAs), 2270, 1541
ciliary movement, 2682 Hybrid antibody-angiogenin protein, 1136 formulations, 2107
postmortem biopsy, 2682 Hybridoma cell, murine, 262 MDI solution formulations
primary cell culture system, 2682 Hybridoma technology, 260, 1132 advantages, 2272
pseudostratified ciliated columnar cells, Hydantoins, 943 disadvantages, 2273
abundance of, 2682 Hydralazine, blood pressure lowering effect, ethanol in, 2272
Human organ system, 1144 573 suspension-based, 2272
Human plasma Hydrastine and sanguinarine, biosynthesis of, MDI suspension formulations, advantages,
enzymatic degradation in, 3013 243 2273
enzymatic hydrolysis, 3011 Hydrate crystal lattice, 1823 propellant, 2272
protein binding, list of, 3034 Hydration-activated drug delivery systems, Hydrofluorocarbon (HFC), 1617
Human platelet-derived growth factor, swellable polymer matrix fabrication, propellant, 997, 2269
recombinant, 1885 1096 nebulization systems, 1283
Human recombinant granulocyte colony Hydraulic compaction simulators, 3209 applications, 2270
stimulating factor (rhG-CSF), Hydride generation technique, 3372 Hydrogel-based microparticles, 2323
2733 for metal analyses, 3369 Hydrogel-modified graphite electrodes,
bioavailability of, 2737 Hydrobromide ophthalmic solution, 389 2030
intracardiac administration, 2737 Hydrocarbon Hydrogel-type dressings
Human respiratory airways, delivery, 2105 chains replacement of, 1052 compositions
Human respiratory tract, 2731 gas phase and liquid phase radiolysis of, agar, 2031
aerodynamic diameter, 2734 3543 chitosan derivatives, 2031
deposition, mechanism of, 2734 and fluorocarbon immiscibility of, 1062 Hyaff, 2031
heterogeneous system, 2734 Hydrocarbon-based ointments, 3258 HypanÕ, 2031
macromolecules absorption of, 2734 Hydrocarbon-based semisolids, 3258 polyacrylamide, 2031
particle deposition in Hydrochloric acid, azeotrope boiling, 3882 polyacrylate, 2031
diffusion, 2734 Hydrochloride salts, 3178 polysaccharide-based hydrogels, 2031
gravitational sedimentation, 2734 Hydrochlorofluorocarbons (HCFCs), 2774 polyurethane, 2031
Human-skin Hydrochlorothiazide, 3437 materials, 2031
in vitro, 3829, 3839 tablets, 911 Hydrogels, 1029, 3264
in vivo model of, 2743 Hydrocolloid adhesive, microstructure of, 2929
Human serum albumin, 1642, 1823, 2674 formulation of, 1032 agarose, 2029
aggregation of, rate of, 1818 superabsorbent, 1032 alginates, 2029
carrier, 1140 adhesive formulation of, 1032 amorphous, 1030
Human simian virus 1 (HSV) vaccine, 1356 for wounds, 1032 as biomaterials, 2021
Human T-cell lymphotrophic virus (HTLV), Hydrocortisone, 2007, 2421 biomedical applications of, 2027
353 an ointment on skin, 3833 as breast prostheses, 2029
Human tissue fluids, 290 penetration, 3821 biosensors preparation, 2030
Human transferrin receptors, 1328 skin permeation, 1314 central nervous system (CNS)
Human tumor solubilization of, 676 regeneration, 2029
cells, 1134 spherical particle distribution, 2389 dental pulp tissue replacement, 2030
xenografts, 1143 ultrasonic nozzle antisolvent precipitation, drug-delivery systems, 2031
Human tumorigenesis, 1417 2389 mandible condylar reconstruction,
Humectants, 995, 2382, 3260 release, 2409 2030
butylated hydroxy toulene (BHT), 996 solubilization of, 3320 ocular implants, 2027
butylated hydroxyanisole (BHA), 996 Hydrodynamic boundary layer, 41 soft contact lenses, 2027
for dermatological preparations, 1554 Hydrodynamic diffusion layer, thickness of, tissue engineering, 2029
examples of, 1554 2670 tissue prosthesis and regeneration,
Humidity Hydrodynamic mechanical stress, 1247 2028–2029
and temperature Hydrodynamic pressure-activated drug wound dressings, 2030
diffusional resistance, 3823 delivery systems, 1094 capsules, synthesis, 1237–1238
ethinyl estradiol, 3823 drug release rate, 1094 carrageenans, 2029
glyceryl trinitrate (GTN), 3823 effective surface area, 1094 chitosan, 2029
norelgestromin (NGMN), 3823 fabrication of, 1094 compression–strain measurements, 2026
serum concentration, 3823 fluid permeability, 1094 cross-linking agents in, 2022
skin barrier function, 3823 hydrodynamic pressure gradient, 1094 divinylbenzene (DVD), 2023

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I73

[Hydrogels] Hydrogenous surfactant, dispersion of, 1050 Hydrophilic molecules, 1246

ethylene glycol dimethacrylate Hydrogen peroxide, 1524 Hydrophilic ointment, 3260
(EGDMA), 2022 biosensors, 2030 Hydrophilic particulate solids
ethylene glycol diglycidyl ether determination, 2030 dispersion, 4124
(EGDGE), 2023 disinfection thermodynamically stable system
glyoxal bis(dially acetal) (GLY), 2023 decomposition, 2208 colloidal silicone dioxide, 4124
N,N0 -methylenebisacrylamide (BIS), on degradation, 2208 microcrystalline cellulose, 4124
2023 with sweet potato peroxidase, 2030 Hydrophilic petrolatum, anhydrous
pentaerythritol triacrylate (PETA), vapor, 2137 absorption base, 3259
2022 Hydrolysis Hydrophilic polymers, 1875
structure of, 2023 acetylsalicylic acid, 316 long chains of, 1878
1,1,1,trimethylolpropane trimethacrylate active ingredient, tablet manufacture, 3673 materials, 644
(TPT), 2022 addition–elimination pathway of, poly-N-vinyl pyrrolidone, 2125
degradation, 1237 2041–2042 polyacrylamide, 2125
disadvantages of, 1238 alkaline, 2041–2042 polyethylene oxide, 2125
divinylbenzene (DVB), 2025 amides, 316 polyhydroxyethyl methacrylate, 2125
drug release from, 2033–2034 compounds behavior of, 2040–2043 polysaccharides, 2125
hydrotropic, 2920 carbamates, 316 Hydrophilic polyoxyethylene, 343
lenses carboxylesterases, 316 Hydrophilic soft contact lenses, 2202
carbenicillin, 1221 definition of, 386 disinfection of, 2206
chloromycetin, 1221 diagnostic test of, 2041 hydrogen peroxide disinfection system for,
gentamicin, 1221 of drug biotransformation, 316 2208
mucoadhesive, 2035 of drugs, 2040 with hydrophilic surfaces, 2203
network parameters of, 2027 of ester, 316, 2040–2041 polymers, ionic nature and non-ionic nature
pH sensitivity of, 2025 hydroxamates, 316 of, 2203
plug, swellable, 1294 kinetic phenomenon, 2046 lens deposits for, 2203
poly(carbamoyl)sulfonate (PCS), 2030 mechanisms of, 2040–2041 lipid deposits, 2203
poly(ethylene glycol) (PEG), 2021 organophosphates, 316 proteinaceous deposits, 2203
poly(vinyl alcohol) (PVA), 2027 reactions, 390, 1613, 1836 tear components, 2203
polyethylene glycol diacrylate, 2030 amidases, 316 Hydrophilic solids, crystal density of,
properties, 1030, 2024 carbonyl derivatives, 2040 3598
bioadhesive, 2035 carboxylic acid derivatives, 2040 Hydrophilic substances, in cattle, 3968
mechanical, 2026–2029 nucleophilic displacements, 2040 Hydrophilic surface characteristics, 1138
swelling, 2024–2025 peptide, 2694 Hydrophilic waxes, microparticles of, 4073
resistance, 2027 phosphatases, 316 Hydrophilizing promoiety, susceptibility of,
rubber elastic behavior of, 2026 phosphoric acid derivatives, 2040 1255
semi-interpenetrating, 2027 reactivity order, 2043 Hydrophobic and hydrophilic polymers,
sheet, 1030 thioesters, 316 combination of, 1291
structure, 1030 Hydrolysis-activated drug delivery systems Hydrophobic anesthetic, 1059
swelling, 2021 fabrication of, 1099 Hydrophobic characteristics, of powders,
synthesis induced degradation, 1099 2978
by cross-linking of polymeric precursors, structure components, 1099 Hydrophobic cyclodextrins, 1258
1238–1239 Hydrolytically labile polyorthoesters, Hydrophobic drugs, 672, 1429
by condensation polymerization, 2021 synthesis of, 186 crystal surface, 41
by free-radical polymerization, 2021 Hydromorphone, 2126 bioavailability of, 3592
monomer in, structures of, 2021 Hydroperoxides, formation of, 2860 encapsulation of, 3579
transdermal release from, 2036 Hydrophile–lipophile balance (HLB), 984, sequestration of, 672
water absorption of, 2021, 2025 1559, 2334, 3605 variety of, 1334
Hydrogenated castor oil, 4069 of emulsifiers, 1560, 3261 Hydrophobic excipients, in tablet, 1740
Hydrogenated cottonseed oil (HCSO), 4075 of surfactants, 1354, 1560, 2214 Hydrophobic interaction chromatography,
Hydrogenated maltodextrin, 2979 Hydrophilic anesthetic, study of, 1059 303, 1189
Hydrogenated soybean phosphatidylcholine Hydrophilic antioxidants, 142 for separation
(HSPC), 1643 Hydrophilic contact lenses, construction, of biological compounds, 531
Hydrogenated vegetable oils, 4066 1221 of proteins, 531
Hydrogen, configuration of, 3083 Hydrophilic drug loaded microspheres, stationary phase in, 531
Hydrogen bond acceptors (HBA), 4014 preparation, by double-emulsion Hydrophobic lubricating agents, 1740
Hydrogen bond donors (HBD), 4014 process, 3590 Hydrophobic membrane filter, 1758
Hydrogen bonding carbonyl group, 827 Hydrophilic drugs, 1242 Hydrophobic molecules, absorption of, 2715
Hydrogen-bonding solvents, 3390 absorption of, 2633 Hydrophobic organic liquids, 1506
Hydrogen bond interactions, 35, 700, 810 fatty base suppositories, 1298 Hydrophobic parameters, calculation of,
Hydrogen and deuterium, differential ability passive diffusion of, 1257 2930
of, 1050 Hydrophilic excipients, 1429 Hydrophobic proteins, 2735
Hydrogen and hydroxide ions, water Hydrophilic liquid substances, 2932 Hydrophobic segments
conductivity determines by, 4040 Hydrophilic matrices, 2407 biodegradable, 2915

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I74 Index

[Hydrophobic segments] Hydroxyls, derivatization of, 676 [Hypersensitivity reaction]

biomedical applications, 2915 Hydroxymelatonin isomers, 2447 type B reactions, 46
pharmaceutical applications, 2915 Hydroxypropyl cellulose (HPC), 1105, 1174, Hypertension
polymer micelles preparation, 2915 1289 treatment of, 2604
Hydrophobic solids, crystal density of, 3598 films, extruded sublingual captopril in, 1077
Hydrophobic sorbents, 531 bioadhesion testing of, 2017 Hypertensive encephalopathy, symptoms of,
Hydrophobic steroids, binding constants of, percent elongation (%E) of, 2015 364
677 tensile strength (TS) of, 2015–2016 Hyperthermia, induced, 1138
Hydrophobic tablet coatings, 2850 Young’s modulus (YM) of, 2015 Hypertonic saline-dextran (HSD), 366
Hydrophobic vent filter, 4044 drug release, 2407 Hypertonic solutions, 3774
Hydropolymer, in wounds, 1029 gelling agents, 1611 Hypnotic agents, absorption of, 1017
Hydroquinone, 2142–2143 films, 2007 Hypnotic capuride, 1017
Hydrosoluble compounds, encapsulation of, explotab in, 2010 Hypoalbuminemia, 1022
1186 glass transition temperatures of, 2009 adverse drug reactions, 49
Hydrostatic head viscometers, with hot-melt extruded, 2010 albumin concentrations in, 3036
Newtonian liquids, 3141 plasticizers role in, 2008 conditions associated with, 49
Hydrotropes. See Hydrotropic polymers. Hydroxypropyl cellulose=wax matrices, liver disease, 49
Hydrotropic copolymers 2010 protein binding, 49
hydrotropic property, 2920 gel, 1885–1886 Hypocalcemia, 2743
optimizing factors, 2920 aqueous preparations of, 1886 treatment of, 3347
synergistic hydrotropic effect, 2920 microviscosity of, 1886 Hypochromic effects, 701
Hydrotropic hydrogels Hydroxypropyl-b-cyclodextrin, 829, 1630, Hypodermic needle, development of,
cross-linked hydrotropic polymers, 2920 3330, 3361 1001
monomers, 2920 manufacturers of, 1642, 1643 Hypoglycemia, 1885, 2908
property, 2920 Hydroxypropyl methylcellulose (HPMC), with insulin, 1078
polymerization, 2920 765, 1105, 1717, 1886 Hypoglycemic effect, 2725, 2743
for solubilization, 2921 advantage of, 3206 Hypothalamus–pituitary–adrenal axis
Hydrotropic monomers capsules versus hard gelatin capsules, suppression, 2633
agents, 2919 3206 Hypothermic effect, use of, 2808
moiety structure, 2919 moisture content of, 3206 Hypotonic solutions
properties, 2919 processability of, 3206 dilution of, 3775
polymerizable monomers, 2919 film system, 669 large-volume infusion of, 3775
Hydrotropic polymer drug release, 2407 water-retention problems, 3775
micelles mixture of, 1288 Hypoxanthine guanine phosphoribosyl
advantages of, 2921 Hydroxypropyl methylcellulose phthalate transferase (HGPRT), 1134
block copolymers, 2921 (HPMCP), 778, 1292 Hypoxic cancer cells, 349
constituents of, 2921 Hydroxypropyl substituent, 672 Hypoxic cardiocytes (HC), 1161, 1162
physical stability, 2921 Hygrometer confocal micrographs of, 1163
solubilization, 2921 dry-bulb, 3884 immunoliposomes, targeting of,
aromatic amide ligands, 2918 wet-bulb, 3884 1163
agents Hygroscopic drug, 767 Hypromellose capsule
for drug solubilization, 2917 Hygroscopicity dimensions, 408
hydrotropes, 2917 of excipients, 38 in vivo performance of, 414–415
water-soluble compounds, 2917 differences in, 3004 manufacture, 406, 408
properties, 2918 of starch, 3479–3480
aggregates, interaction of, 1059 Hyoscyamine, 76
chemical structures, 2918 Hyperbilirubinemia, 1018, 2639 Ibuprofen, 996, 1125, 1861, 2154
coabsorption of, 2918 HyperDSCTM, 396–397 absorption rates of, 426
for hydrophobic drugs, 2918 amorphous content detection, in biotransformation of, 311
hydrotropic polymers, 2917 amorphous=crystalline lactose card pattern of, 4105
solubilization, 2918 blends, 396 crystals
water solubility of, 2918 calibration, 396–397 preparation of, 829
Hydroxy alkylation, 684 enthalpy standards, 396 in Dolgit Mikrogel, 1126
Hydroxyallyl cellulose, drug release, 2407 measurement of, heat of fusion, 396 enantiomer–enantiomer interactions of,
Hydroxybenzoate esters, 1006 sample analysis, 396 2156
Hydroxyethyl acrylate, 2930 Hyperglycemia, 2908 enantiomeric composition of, 3436
Hydroxyethyl cellulose, 2125 Hypericum perforatum, 75–76 isomer conversion, 321
Hydroxyethyl methacrylate (HEMA) Hyperlipidemia, 1018 NMR spectrum of, 3445
application, 1221 treatment, 2923, 3348 permeability of, 1126
based lenses, 1221 Hypersensitivity reaction, 46 Ibuprofen enantiomers
copolymer composition, 1221 gastrointestinal (GI) tract, 47 lysinate, 1125
polymers, 1221 immune-based reactions, 46 stereospecific oxidation, 320
Hydroxyethyl starch (HES), 1822–1823, 1828 in pharmaceutical dosage forms, 47 tablets, 925
Hydroxylamine, dehydration of, 314 reduction of, 1331 Ibuprofen-wax, 4073

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I75

Ice–vapor boundary, 1807 [Immunoassay] Immunodiffusion (Ouchterlony) discs,

crystallization, 1808 for biomarkers, 1571 double, 1418
rate of, 1811 cross-reactivity of antibody, 1572 Immunogenic antigen production, in plants,
crystals definition, 1571 3911
growth of, 1808 implementation of, 1571 Immunogenic foreign epitopes, 3910
size of, 1811 in vivo biological properties, 1571 Immunogenic moieties, inclusion of, 1141
growth physiological measurement, 1571 Immunogenic reactions, 1138
rate of, 1811 biomedical analysis, 202 Immunogens, subunit vaccines, 3913
nucleation, 1809 chemiluminescence, 204, 2057 Immunoglobulin, 2048
Ice, vapor pressure of, 3893 clinical development, 1572 fraction, 1132
ICH. See International Conference on competitive binding analysis, 202 fragmentation of, 1144
Harmonization. chemical properties, 202 low pH incubation of, 3999
Ichthyosis, 3821 critical binding reagent, 1572 process of, 2724
ICP. See Inductively coupled plasma. cross-reacting metabolites, 1573 safety of, 3999
IDENT computer program, development of, physical properties, 202 synthesis, 1132
2766 spectral properties, 203 Immunohistochemical staining, 1328
Idiopathic cardiomyopathy, 1156 detection system end-point, 1576 Immunoliposome
Idiosyncratic reaction, 46 diagnostic methodologies, 202 advantage of, 1140
IDR. See Intrinsic dissolution rate. drug analysis, 202 antimyosin, 1159
Ifetroban, synthesis of, 2995 electrochemical, 1526–1528, 2058 applications of, 1133, 1140
IgG-mediated immune complex reaction, 46 enzyme, 203 in vivo research on, 1142
IgG humoral-mediated immunity, 1421 flow injection, 1577 rhodamine labeled, 1162
IgG or IgM-mediated cytotoxic reaction, 46 fluorescence, 2054 system, 1142
IgM isotype antibody, 1158 fluorescence polarization, 2055 for transfection, 1165
Ileal enterocytes, microvilli of, 2715 heterogeneous amperometric, 1527 Immunomicrospheres, 1143
Ileocecal junction, 1244 heterogeneous electrochemical enzyme, efficacy of, 1143
Image analysis software, automated, 859 1527, 1528 Immunomodulation
Image-analysis techniques, 2586 heterogeneous, 203, 2048 costimulation, 3922–3923
Image detectors, 3399 fluorescent labels, 203 cytokines, 3923
Imines, hydrolysis of, 2045 radioimmunoassay, 203 treatment
2-Iminothiolane, 1135 homogeneous, 203 chronic infectious diseases, 3922
reagents, 1136 excitation wavelength, 203 neoplastic diseases, 3922
Imipramine, 575 fluorescent labels, 203 Immunomodulator, synthetic, 674
Imitanib, 265 fluorescent signal, 203 Immunoradiometric assay (IRMA),
Immediate release capsule formulations, homogeneous amperometric, 1528 1570
2407 immunoglobulin G molecules of (IgG), Immunotherapy, 1149
Immersional wetting, 3585 2048 Immunotoxic events, 1616
Immobilized antibody, 1567 liposome, 2059 Immunotoxicology, 1420
Immobilized antigen, 1567 low-molecular weight xenobiotics, Immunotoxins, 1139, 1150
Immobilized-enzyme reactors (IERs), 1525 1573 immunogenicity of, 1139
Immune effector functions, 1137 luminescence, 203 therapy, 1139
Immune responses chemiluminescence, 203 tumor associated antigen, 1150
division, 3914 fluorescence, 203 Impeller pumps, fluid flow, 3869
polarization of, 3914 for macromolecules, 1569, 1573 Impeller torque, 4081–4081
Immune-stimulating complexes (ISCOM), matrix effects for, 1573 Impurity profile, control of, 2830
3914, 3916, 3595 for pharmacological markers, 1571 In-capÕ, 410
adjuvanted vaccine, 3916 physiological measurement, 1571 Increased pulmonary residual volume
Immune system potentiometric, 1527 (IPRV), 348
adaptive immune system, 3914 precipitation reagents, 2049 Indicator electrode potential, 1509
division, 3914 pre-clinical development, 1572 Indirect Fourier transform (IFT), analysis of,
innate immune system, 3914 predefined criteria for, 1573 1054
Immunization, non-invasive method of, protocols Indium, thermal lag for correction, 3727
2746 radiolabeled Indomethacin farnesil, 580, 3348
Immunoanalytical methods antibody, 2050 amorphous form of, 1797
Immunoassay antigen, 2049 biotransformation of, 311
advantages of, 2048 selectivity and sensitivity, 202 calcium salt formation of, 3181
for antibodies, 1569 separation-free homogeneous, 203 delivery of, 1237
validation of, 1574 advantages of, 203 across rat skin by sonophoresis, 3832
antibody–antigen reaction, 1573 drugs for, 203 Indomethacin-loaded nanocapsules, 1195
antibody–protein conjugates in, 1572 structural properties, 1574 complexes, 700
automation of, 1577 for surrogate markers, 1571 preparation of, 700
bifunctional glycoproteins, 2048 Immunochemical reaction, 1526 suppositories, 924
bioanalytical method validation, 1572 Immunoconjugate, 1133 Inductively coupled plasma (ICP), 3367
in biological matrices, 2048 preparation of, 1136 advantage of, 3373

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I76 Index

[Inductively coupled plasma (ICP)] [Infrared spectroscopy] Inhalers

argon plasma flame, 3373 in mid-IR region breath-activated device, 1923
spectrophotometry, 3373 fingerprint region, 3415 diskhaler, 1923
Inductively coupled plasma-atomic emission functional-group region, 3415 in geriatric patients, 1922
spectrometry (ICP-AES), 1759 matrix-isolation technique, 3417 metered-dose inhaler (MDI), 1923
Inductively coupled plasma–mass of polyatomic molecules, 3407 as nebulizer, 1923
spectrophotometry (ICP–MS), qualitative analysis, 3414 as turbuhaler, 1923
3373 quantitative analysis, 3416 Inhaler’s dispersion efficiency, improvement
Industrial pharmaceutical technology (IPT), base-line measurement method, 3416 of, 1431
3697 of multiple components, 3416 Injections
Infant respiratory distress syndrome (IRDS), Infrared-transmitting materials, properties, active ingredient, 1266
1128 3412 administering pharmaceutical preparations,
Infectious recombinant viruses, 645 Infrared tunnels, disadvantages of, 959
Inferior vena cava, 2717 3515 antimicrobial preservative agents,
Inflamed gum tissues, 899 Infrared window materials, transmission 1267
Inflammatory bowel disease, 1242, 1254 ranges for, 3415 concentrated liquids, 1266
Inflammatory cell infiltration, 686 InfusaidÕ constituents of, 1266
Inflammatory diseases, incidence of, 3036 implantable infusion pump, 1095 Inorganic materials, 3678, 3680
Inflammatory reactions, treatment of, vapor pressure-activated drug-delivery consolidation mechanism, 3680
1300 system, 1097 from dibasic dihydrate, 3680
Inflammatory response syndrome, 369 Infused oils. See Oils, infused. as direct compression diluents, 3678
Influenza virus, 3918 Infusion control device (ICD), 1010 pharmaceutical excipient, 3680
Informed Consent (IC) forms, 1926 Infusion gentamicin, 2640 Insecticides, 1615
InfraGold, 3377 Infusion pumps and devices, 1010 Insensitive Nuclei Enhanced by Polarization
Infrared absorption and Raman scattering, Infusion therapy, 1002 Transfer (INEPT), 3445
2942 Inhalants, constituents, 958 Insessia, 959
Infrared absorption bands, structure and, Inhalation aerosols, gamma scintigraphic Installation Qualification (IQ)
3415 imaging of, 3094 cleaning process, 1587
Infrared analysis Inhalation devices, 21 CIP skid, 1587
gaseous samples, 3412 Inhalation droplet spray plume, by monitoring equipment, 1587
variable path gas cell in, 3413 TouchSpray, 3856, 3857 spray device, 1587
liquid samples, 3413 Inhalation drug delivery devices cleaning validation, 1587
sample cells in, 3413 categorization Instrumentation amplifier
low-boiling liquids, 3412 high frequency oscillating devices, antialiasing filter, 3687
solids, 3413 2104–2105 signal conditioning, 3687
film, 3413 pressure-driven devices, 2095 accuracy, 3687
mull, 3414 dry powder inhalers (DPIs), 2094 calibration, shunt, 3687
pressed disk, 3414 metered dose inhalers (MDIs), 2094 dynamic range transducer, 3687
solution, 3413 nebulizers, 2094 excitation, 3687
Infrared analyzers, 3410, 3417 Inhalation therapy, 1203 full scale (FS), 3687
applications, 3417 driving factors hysteresis, 3688
Infrared and viscometric measurement, 1057 aerosol generation technology, 2092 measurement errors, 3688
Infrared heaters, 3515 biotechnology advances, 2092 nonlinearity, 3688
Infrared region, absorbance in, 2941 patient breathing patterns, 2094 precision, 3687
Infrared spectrometer inspiratory flow rate, 2725 repeatability, 3688
components, 3407 Inhalation toxicity testing, 2270 resolution, 3687
data-handling system of, 3411 Inhalation toxicology potential, 2774 sensitivity, 3687
detectors in, 3408 Inhalations, medicinal agents administration, traceability, 3688
quantum, 3409 958 on tablet press, 3686
thermal, 3409 Inhalations, dry. See Inhalants. Insulin
dispersive, 3407, 3409 Inhale therapeutic systems, 1432 absorption of, 2705, 2726, 2632
monochromator in, 3408 and Alkermes, 2090 to beagle dogs, 1078
interferometer in, 3408 pulmonary drug delivery technology, biologic activity of, 2727
monochromator in, 3408 1279 buccal formulations of, 1078
non dispersive, 3407, 3410 Inhaleables chemical structure of, 704
interferometer in, 3408 advantages of, 1279 crystallography of, 704
optical components, 3410 definition of, 1279 delivery of, 185
radiation sources for, 3407–3408 dispersible formulations, 1279 injection of, 1888
sampling techniques, 3412 drug aerosol delivery devices, 1279 iontophoresis, 2749
transmittance, 3412 gastrointestinal tract problems, 1279 lyophilized, 286
Infrared spectroscopy, 699, 3405, 3406 non-invasive method, 1279 in plasma concentration of, 1304
applications, 3417 respiratory tract diseases, 1279 pulmonary delivery system, 2705
instrumentation, 3407 treatment of, 1279 in rabbits, 1078
interpretation of, 3415 systemic diseases, treatment of, 1279 recombinant human, 260

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I77

[Insulin] Interfacial free energy, 3583 International Conference on Harmonization

rectal absorption of, 2728 Interfacial tension (ICH)–Expert Working Group on
release, 2033, 2409 advantages of, 811 Drug Quality (EWG-Q) guidelines,
secretion of, 2696 changes in, 1306 1963
in treatment of glycemia, 1194 intermolecular forces, 811 International Conference on Harmonization
zinc crystals, 1798 lowering of, 1254 (ICH) (1991–1997), 439–440
IntalÕ, 2107 macroscopic determination, 811 International Conference on Harmonization
Integrated automatic sampling plant, 2966 solute–cosolvent, 811 (ICH) guidances
Integrated iontophoretic devices, 2746 solute–solvent, 811 drug impurities
Integrins, definition of, 1328 solute–water, 811 products, 3798
Integrins signals cell proliferation, 1328 solvent, 810 substances, 3797–3798
Integrity test measurements Interferometer, 3434 residual solvents impurities for, 3797
polymer membrane, 1755 components of, 3408 International Conference on Harmonization
pore size of membrane, 1755 Interferon-b Residual Solvent Guidance,
surface area of filter, 1755 cytokine therapy, 3923 1656–1657
wetting fluid, 1755 with multiple sclerosis, 3923 classes, 1656
Intestinal absorption, 3008 Interferon-g (IFN-g), oxidation of for pharmaceutical excipients, 1656
barriers, 2716, 2717 recombinant, 285 International Diffuse Reflectance Conference,
enhancers, 16–17 Interferons, 269, 271 3434
Intestinal apical cell membrane, 2721 indications for, 269 International Federation of Pharmaceutical
Intestinal blood flow, 1258, 2716 Interleukins, 271–272 Manufacturers Association
Intestinal cell membrane, 2721 InterlinkÕ system, 1009 (IFPMA), 1961
Intestinal digestive enzymes, 1246 Intermicellar interaction, 1055, 1059 International Generic Pharmaceutical
Intestinal epithelial barrier, 1246 intradermal, 1215 Alliance (IGPA), 4102
Intestinal epithelial cells, 1244, 2718, 2720 oils for, 1267 International harmonization
drug transport across, 1245 parenterals, 959 with JP (Japan pharmacopeias), 2834
Intestinal epithelium, 1177 sterile solutions, 959 process of, 2837
membrane permeability of, 1258 suspensions, 959 programs for, 2852
surface of, 1242 Intermolecular attraction forces, weak, 38 USP(U.S. pharmacopeias), 2834
Intestinal fluids, drugs insoluble in, 1253 Intermolecular interactions, 616–619 International Nonproprietary Nomenclature
Intestinal lumen, 1255, 2713 in benzamide crystal, 617 (INN) Committee, 1891
drugs in, 2718 types of, 86 International Normalized Ratio (INR),
exposed in, 1301 International Association for Food 1906
Intestinal lymphatics, 1244 Protection (IAFP), 2237 International Organization for
Intestinal membrane, 2714 International Association of Milk, Food and Standardization (ISO), 3525, 3940
enterocytes of, 2722 Environmental Sanitarians of health care products, 2290
diffusion across, 1245 (IAMFES), 2237 voluntary standards of, 2290
permeability of, 2720 International Centre for Diffraction Data International Pharmaceutical Aerosol
Intestinal metabolism, 576, 578 (ICDD), 4104 Consortium (IPAC), 2106
first-pass, 1247 International Committee on Radiation International Pharmaceutical Aerosol
Intestinal mucosa, 347, 942, 1077, 2870 Protection (ICRP), 3093 Consortium for Regulation and
apical membrane of, 1246 International comparator products, list of, Science (IPAC-RS), 1698
barrier function of, 1248 4100 International Pharmaceutical Aerosol
metabolic activity of, 2716 International Conference on Harmonization Consortium for Toxicity Testing
Intestinal mucosal cells, 216 (ICH) (IPACT), 2270
Intestinal mucosal surface, 1245 chemical entities, 1962 International Pharmaceutical Excipient
Intestinal peptide transporter, 2722 on drug quality, 1960, 1961 Council’s (IPEC), 1619, 1632, 2771,
Intestinal permeability, 1256 guidelines, 1407 3642
effective, 1246 adverse drug reactions (ADRs), 2497 chemical excipients, 1632
low, 1244 adverse events (AEs), 2497 classifications, 1632
Intestinal secretions, 24, 2870 serious adverse events (SAEs), 2497 in Europe, 1656
Intestinal surgery, 1017 guideline on stability testing of new drug food processing firms, 1656
Intestinal transit rate, 2870 substances and products, 1685, 1686 in Japan, 1656
Intestinal transmembrane protein, 213 impurities, 1962 in United States, 1656
Intestine methods validation defined by, 92 International pharmacopeia, 2856
large, 2716 pharmaceuticals for human use, 1781 scope of, 2857
small, absorptive surface area of, 1246 regulatory definitions and requirements, International Society for Pharmaceutical
Intestine, absorptive surface area of, 1242 92 Engineering (ISPE), 1, 2290
IntestinolÕ, 1254 S7B Safety Pharmacology guideline, 2196 International Union of Pure and Applied
Interactive voice response (IVR) systems, 734 safety data management by, 2500 Chemistry (IUPAC), 445, 1548
Intercellular binding force, 2665, 2927 veterinary products, 1781 in absorption measurement, 3461–3462
Interdigestive myoelectric cycle, definition of, efficacy, 1788 Interpharmacopeial Open Conference on
1244 quality, 1788 Standards for Excipients, 1960
Interfacial energy, 35 safety, 1788 Intraarterial injections, 2991

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I78 Index

Intracellular bacteria Intraurethral pressure (IUP), model of, 2809 [Ion exchange]
Listeria monocytogenes, 3923 Intrauterine devices (IUD), 1356 synthetic resins
Intracellular drug Intravascular persistence, 339 on anions and cations, 4040
metabolism, 2716 Intravascular retention, 354 Ion-exchange and ion chromatography,
release, 1333 Intravenous administration, 2639, 3027 530–531
Intra-celluar enzymes, 2716 of drug, 1299, 2641 mobile phase in, 530
Intracellular infections of nanoparticles, 1189 stationary phases in, 530
for drug delivery, 1190 types of, 1003, 2642 anion exchange resins in, 531
treatment of, 1190 Intravenous atropine sulfate, 2821 cation exchange resins in, 531
Intracellular Leishmania donovani, with Intravenous dosing, 939 polystyrene divinylbenzene copolymer
nanosphere, 1191 Intravenous drug solutions, 218 resins, 531
Intracellular lipids, 1257 Intravenous epinephrine (EPI), effect of, 944 Ion exchange chromatography (IEC), 303
Intracellular membrane-coating granules, Intravenous immunoglobulin (IVIg), 3999 deionizers, 4041
1311 Intravenous infusion therapy, 3052 glycoproteins analyses, 531
Intracellular metabolic machinery, 1301 Intravenous injection, 1143 peptides analyses, 531
Intracellular metabolic processes, 705 advantages, 3954 proteins analyses, 531
Intra-Peyer’s patch immunization, intestinal of antimicrobial agents, 3954 resins, 531
immunization by, 3918 disadvantage, 3954 system
Intracellular pathogens and tumors, 3915 drug administration in animals, 3954 types of, 4041
Intracellular proteins, coagulation of, 329 ear vein, 3954 Ion-exchange polymers, 2122
Intracellular reactions, 325 formulation of, 1669 Ion-exchange chromatography, stationary
Intracellular tumor delivery, composition of, irritant solutions, 3954 phases, 530
1142 jugular vein, 3954 Ionic crystals
Intracellular water (ICW), 2634 multiple, 1145 color centers, 3545
Intracytoplasmic DNA delivery, 1164 in neonatal foals, 3954 electronic localization in, 3545
Intraepithelial lymphocytes, 2714 Intravenous therapy and pyrogens, 3052 Ionic equilibria
IntrajectÕ device, 1213, 1216 Intraventricular ventricular hemorrhages, acidic and basic substances, 385–386
IntralipidÕ, 984, 3362 2641 buffer systems, 387–388
Intramammary preparations, 3981 Intrinsic dissolution rate (IDR), 909, 910, conjugate acids and bases, 386–387
Eazi-Breed CIDRÕ, 3981 2943 Ionic exchange equilibria, 1506
intramammary products, 3981 Intrinsic factor vitamin B12 complex, 2715 Ionic gelation
PirsueÕ, 3981 Intrinsic proteins applications, 2317
Intramolecular catalysis, 3011 cell membrane, 25 polyelectrolytes cross-linking in, 2317
Intramolecular cyclization–elimination, 3009 globular proteins, 25 polymer systems in
enzymatic reaction of, 3008 IntrocanÕ, 1009 carboxymethylcellulose=aluminum, 2317
mechanism of, 3011 Inulin elimination chitosan=triphosphate, 2317
Intramolecular hydrogen bonding, 2936 glomerular filtration, 3965 gelan gum=calcium, 2317
Intramuscular absorption, 2631–2632 Inulinase, 2035 k-carrageenan=potassium, 2317
Intramuscular administration, 2645 Inunctions, 959 pectin=calcium, 2317
Intramuscular hemorrhage, 2632 Inunctum. See Inunctions. polyphosphazene=calcium, 2317
Intramuscular injection, 209 Investigational Exemption for a New Drug Ion-interaction chromatography, 530
amoxicillin trihydrate in neck of pigs, 3956 (IND), 1406, 2468, 2469 Ionizable cyclodextrins, 1258
of antibiotic, 1307 Investigational new rug (IND), 1780, 1834 Ionizable polymers, types of, 1254
ceftiofur sodium, 3954 clinical studies, 1783 Ionization
disadvantages, 3952 and new drug applications (NDA), constants determination of, 390
drug absorption from, 3952 1783 degree of, 2988
drug administration in animals, 3952 non-clinical studies, 1783 Ionizing radiation
ketamine hydrochloride, 3955 Investigational new drug applications absorption of, 3549
in lateral neck, 3952 (INDAs), 2993 biological actions of, 3547
of parenteral ampicillin formulations, 3956 and stability information systems, 733 biological inactivation of, 3549–3550
quadriceps muscle mass, 3952 Iodine(tri-iodide)-iodide couple, 1505 micro-organism killing by, 3550
Intramuscular pentagastrin, treatment of, Ion-activated drug delivery systems, 1098 biological significance, molecules of, 3547
2821 air-suspension coating technique, 1099 on biological systems, 3540
Intranasal systemic delivery, applications, diagrammatic illustration of, 1099 characteristic of, 3540
2684 gastrointestinal fluid, 1098 chemical protection from, 3546–3547
Intranasal vaccination, 3918 structural components, 1099 application of, 3548
Intraocular lens (IOL), 2028 IonaminÕ, 1771 energy and charge transfer, 3547
Intraocular pressure (IOP), 944 Ion channels, in biological membranes, 26 molecular complexes, 3547
Intraoral jet-injection technique, DNA Ion chromatography, for separation and scavenging intermediates, 3547
vaccine delivery, 3916 conductivity detection, 531 DNA by
Intraosseous administration, 2632 refractive index detectors in, 534 exposure dose, 3550
Intrathecal administration, 3059 Ion exchange, 4040 in food processing, 3551
Intratracheal administration, 2732, 2733 and membrane separation technologies on living cells, 3549
Intratumoral distribution, 1330 combination of, 4041 on molecules, 3547

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I79

[Ionizing radiation] Iontophoresis electrode technology [Irradiation]

mutagenesis by, 3549 Ag=AgCl electrode system, 3848 aqueous solution, of cysteine, 3547
for pharamaceutical sterilization, 3552 electrochemical products, 3849 aromatic compounds, 3544
physical and chemical actions of, 3540 electrochemical reaction, 3848 carbohydrates, 3547
on solute molecules, 3543 ion-exchange resin, 3849 coenzymes, 3548
in sterile pharmaceuticals preparation, 3551 ion-selective membrane, 3849 disulfides, 3547
sterilization non-sacrificial (catalytic) electrodes, 3848 DNA, 3548
application of, 3552 transdermal iontophoretic transport, 3848 enzymes, 3548
of drugs, 3551 Iontophoretic drug delivery systems (IDDS) frozen alcohols, 3544
target theory, 3547, 3549 components ice, 3546
Ion-pair chromatography, 530 adhesive laminate, 2129 ionic crystals, 3545
Ion-product constant, for water, 3754 bottom housing, 2129 of liquids, 3542
Ion-selection electrode technology, 1762 electrodes, 2129 molecular crystals, 3545
Ion-selective carriers, class of, 1507 hydrogels, 2129 nucleic acids, 3548
Ion-selective electrodes (ISEs), 1519–1520 printed circuit board assembly, 2129 DNA and RNA, 3548
analyzers, 1760 electronic components of, 2128 para-aminobenzoic acid, 3548
calibration graphs of, 1509 control circuitry, 2128 peptides, 3547
techniques, 1512 power source, 2128 polymeric materials, 3543
Ion-selective membrane electrode, 1506 fentanyl delivery from, 2129 polysaccharides, 3547
IonsysTM E-TRANSÕ system, Alza Corp., fentanyl hydrochloride in, 2125 proteins, 3547–3548
CA, 3848 formulations of respiratory proteins, 3548
Ion trap mass spectrometer drug salt, 2125 riboflavin, 3548
in metabolite characterization, 2264 excipients, 2126 saturated aliphatic compounds, 3544
metabolic alteration, 2264 formulation pH, 2127 silica gel, 3546
sequential product, 2264 solvent, 2124 sources
Iontophoresis, 2119, 2746 luteinizing hormone-releasing hormone selection of
applications delivery from, 2131 ICH guidelines, 2863
cystic fibrosis diagnosis, 2119 operation, 2122 UVA, 2862, 2863
dexamethasone administration, 2119 patient-activated, 2125 UVB, 2862, 2863
glucose removal from body, 2119 treatments, 2130 Visible (VIS) light, 2862, 2863
hypo- and hyperglycemia detection, Iontophoretic electrodes surface catalysts, 3546
2119 assemblies systems, protection of, 3546
lidocaine administration, 2119 counter electrode, 2121 thiol-containing amino acids, 3547
therapeutic delivery across skin, 2119 donor electrode, 2121 vitamins, 3548
treating eye, ear, nose, and mouth, 2119 consumable or sacrificial type water, redox properties of, 3543
delivery efficiency in, 2121 capacity, 2122 Irradiators
devices non-consumable or inert type, 2122 continuous carrier, 749
chemistry-related issues, 3848 disadvantages, 2122 radioisotope, 749
components, 2119 polarity, 2121 Irreversible process
design of, 3848 International Pharmaceutical Excipient coalescence, 1555
in iontophoretic transport, 3848 Council’s (IPEC)-America safety Ostwald ripening, 1555
on skin, 2119–2120 testing guidelines, 1659 Irrigating solutions. See Solutions, irrigating.
electrodes, 2121 excieptiens testing in humans, 1662 Irritant
consumable or sacrificial type, 2122 for pharmaceutical excipients, 1656 classified as, adverse skin, 1315
counter electrode, 2121 International Pharmaceutical Excipient contact dermatitis, 1315
donor electrode, 2121 Council’s (IPEC)-Europe excipient Irritation
medical potentiometric, 2121 testing guidelines, 1660 muscle and vein, 1410
non-consumable or inner type, 2122 Intraocular pressure(IOP) reduction, 1222 ocular and dermal, 1409–1410
silver=silver chloride redox couple, 2123 Ipratropium bromide variables influencing, 1425
materials and excipients in, 2119 anhydrous, 4056 Ischemic acidosis, 358
migration blocking in, 2122 water Ischemic brain injury, 364
of drugs, 2119 desorption isotherms for, 4056 Islet cells, 1900
skin permeation physical methods of, 1311 vapor sorption for, 4056 Islets of Langerhans, immobilized,
transport mechanism Ipriflavone, 2441 2336
electromigration, 2119–2120 IR spectra, measurement of, 863 ISO (International Organization for
electroosmosis, 2119–2120 Irgafos 168 Standardization), 1404
electroporation, 2119–2120 chemical ingredient, 1695 Isobutene, in aerosol consumer, 2270
treatment, 2128 in extractables=leachables, 1695 Isocratic chromatography, 98
Iontophoresis-activated drug delivery packaging component, 1695 Isocratic elution, 526
systems, 1096 Iron-catalyzed oxidation, 1825 disadvantages of, 527
components of, 1096 Irradiation Isoelectric focusing (IEF), 304
development of, 1096 aliphatic compounds, 3544 Isoenzyme system, 71
passive diffusion, 1096 alkyl halides, 3544 Isoflavones
skin permeation rate, 1096 amino acids, 3547 sources, 2442

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
I80 Index

[Isoflavones] Isomerization=racemization, 286 Jake leg paralysis. See Jamaica ginger

for women’s health, 2442 Isomerization to fructose, 1614 paralysis.
Isoflavonoids, 237 Isomers, 2142 Jamaica ginger
biosynthesis of, 236 binding differences between, 2153 with castor oil, 1406
Isolation technology, in aseptic processing, definition, 2142 paralysis, 1406
137 geometric, 2146 Japan Pharmaceutical Excipients Council,
Isolator(s) of ephedrine, 2145 2837
alpha=beta port in, 2139 types Japanese Ministry of Health and Welfare
applications, in powder processing systems, configurational, 2142 (JMHW) evaluation system, 2771
2135 conformational, 2142 Japanese Pharmaceutical Excipients, 660, 691,
large-scale aseptic production, constitutional, 2142 1633
2136–2137 Isoniazid non-official, 1632, 1639–1640
small-scale manipulations, 2136 elimination of, 1018 official, 1632, 1634–1638
in sterility testing, 2134–2135 release, 2033 Japanese Pharmaceutical Manufacturers’
in subdivision and potent-drug Isopentenyl pyrophosphate Associations of Tokyo, 2837
dispensing, 2134 compounds derived from, 239–246 Japanese Pharmacopoeia (JP), 660, 1623,
in syringe filling, 2137 Isoperibol calorimeters, 402, 3738 1633
cleaning and sanitization of, 2141 crystallinity of drug substances by, 3738 Japanese Pharmacopoeial Forum, 1964, 2837
critical task for, 2133 Isoprenaline, 991, 2862 Japanese Society of Hospital Pharmacists,
definition of, 2133 Isoprenaline sulphate, 991 2837
design Isoprene Jellies, 995
and construction, 2134 elastomer, 1466 mongraphs in USP, 1876
considerations for environment, 2141 rubber, 1466 See Gels.
features, 2133 Isoprene-derived compounds, 236 single-phase, 1875
flexible-film, 2133 Isopropanolamine salts, 941 sodium hyaluronate, 1884
glove and gauntlet materials in, 2138 Isopropyl alcohol, 2478, 2942 Jenike shear cell, 3287–3288
half-suit, 2138 Isopropyl myristate, 14, 2980 advantage and disadvantage of, 3288
in health care and life science industries, in cosmetic preparation, 993 to measure cohesive powders, 3287
2133 permeability coefficient, 1313 pharmaceutical powders by,
for health care regulatory demands, 2134 Isopropyl palmitate, in cosmetic preparation, characterization of, 3287
leakage-rate tests, 2138 993 Jet nebulizers
leak-induction test, 2138 Isoproterenol in airway diseases, 2095
leakage-rate measurement, 2138 cardiotoxic effects of, 149 consequences, 2095
pressure decay rate evaluation, 2138 sulfaction of, 1247 droplet formation by, 2095–2096
tracer gas leak-rate detection, 2138 Isoquinoline alkaloids, 248 drug concentration effects in, 2096
in manufacture of sterile products, 2134 Isosbestic point, 3461 effect of physical properties in, 2096
in manufacturing R&D, and QC sectors, Isosorbide-5-mononitrate, 1076, 1234, Pari LC, 2096
2134 1253 physicochemical properties of, 2096
for pharmaceutical application, 2133 for angina prophylaxis, 1076 temperature effects in, 2097–2098
positive-pressure, 2134 Isosorbide dinitrate, 1076, 1253 Johanson Hang-up Indicizer system,
potent drug-dispensing, 2135 antianginal action of, 310 3290
rigid-wall, 2134 Isothermal calorimetry (ITC), 306 pharmaceutical powders by,
sleeve=glove, 2138 Isothermal microcalorimetry, 403, 3302 characterization of, 3290
technical considerations for design, of amorphous component of lactose, Joint Commission on Accreditation of Health
manufacture, and testing of 3302 Care Organizations (JCAHO)
airborne cleanliness classification, 2140 Isothermal titration calorimetry, 1884 Standards, 46, 2188
airflow configuration, 2140 Isotonicity, 1272 Joint Expert Committee on Food Additives
barrier-integrity characteristics, in biological cell, 1272 (JECFA), 660, 691
2137–2138 solution tonicity, 1272 Joint health, 2431
HEPA filtration, 2140 Isotope-coded affinity tagging (ICAT) diseases
internal pressurization, 2139–2140 technology, 3045 osteoarthritis, 2431, 2435
manipulation technique, 2138 Isotopes, of hydrogen, 3082 rheumatoid arthritis, 2431, 2435
open aperture protection, 2140–2141 Isotopic dilution analysis, 3089 nutraceuticals in, 2431
transfer techniques, 2138–2139 advantage of, 3090 treatment with, 2435
technical guidelines and standards, inorganic trace elements by chondroitin, 2437
2133–2134 determination of, 3090 dimethyl sulfoxide (DMSO), 2437
walls or envelopes, 2133 Isotoxic concentrations, 2988 glucosamine, 2436
Isomeric copolymers, 1064 Isotretinoin, 3348 S-adenosyl methionine (SAMe), 2437
Isomerism, 2142 Isozymes, multitude of, 2635 Joint Interpreting and Conference Service
and analytical methodology, 2156 Itraconazole, 616, 620, 3350, 3361 (JICS), 1597
conformational, 2148 oral bioavailability of, 3350 J-Tip needle-free injector, 1217
geometric, 2146 Ivermectin (IVM) Jugular vein, collection of blood samples,
pharmacological aspects, 2149 kinetic parameters for, 3955 3954
background and definitions, 2149 topical solution of, 3971 Juleps, 960

Volume 1: Pages 1–670; Volume 2: Pages 671–1434; Volume 3: Pages 1435–2118; Volume 4: Pages 2119–2828;
Volume 5: Pages 2829–3482; Volume 6: Pages 3483–4128.
Index I81

KalrezÕ, 2237 Kinetic turbidimetric [Laboratory Information Management

Kaolin, 989 assay, 3058 System (LIMS)]
Karl–Fischer reagent, 3759 and chromogenic assays, 3059 organizational issues, 2169
Karl Fischer titration KiraÕ, 75 pharmaceutical standards, 2167
advantages and disadvantages of, 2377 Kjeldahl method quality of, 2166
for cellulose, 2377 inorganic nitrates and nitrites, 3756 querying for, 2170
for dicalcium phosphate, 2377 Klebsiella species, 2479 securing funding, 2166
for lactose, 2377 Knauer membrane osmometers, 3776 standard requirements, 2167
moisture content determination by, 2377 KoateÕ, 4002, 4008 static data, 2165
reagent for water, 2377 Kohonen networks, 2402 system configuration, 2164
of water, 2377 Kollidon CLÕ, 3556 template data, 2165
Karposi’s sarcoma-associated herpevirus Konseal apparatus types of standards for, 2166
cloning of, 3123 components of, 951 validation
Kava (Piper methysticum), 74 saucer shaped rice-flour, 951 of systems for 4, 6
KBr pellet absorbance spectrum, 3379 Konseals, 960 time, 2166, 2169
KBr pellet technique Kozeny–Carmen equation, 2590 vendor audit, 2167, 2169
of caffeine, 3379 Kronig–Kramers transform, 449 LabrafilÕ, 3335
for mid-infrared measurement, 3378 KryotabTM, ODT technologies in LabrasolÕ, 3348
KBr powder development, 1111 Lactate dehydrogenase (LDH), 370, 1430
hygroscopic nature of, 3382 Kubelka–Munk (K–M) function, 3377 Lactic acid, formation of, 1615
reflectance of, 3382 linearity of, 3381 Lactide-glycolide copolymers, 160
k-carrageenan gel, 1884 Kupffer cells, 369, 1191 Lactitol, 3681
Kefauver–Harris Amendment, 561 Kuru disease in human, 1642 as tablet diluent, 3681
result of, 2904 Lactobacilli, 1341
Kefauver–Harris Drug Amendments, Lactobacillus acidophilus, 2257
1407, 1941, 3073 Labeled antibody radioimmunoassays, Lactose, 998, 1233, 3681
Kefauver-Harris amendments, 2419 2050 advantage of, 1615
Kelvin effect, 1556 Labeled antigen radioimmunoassays, 2049 amorphous, 3230
Kelvin model of, viscous materials, analytical reagents, 2049 component of, 3302
3135 radioactivity measurement, 2049 anhydrous, 3681
Keratinization, 2665 Label-driven food-effect assessments, 2821 ball-shaped spray-dried, 3230
degree of, 2671 Labile pharmaceuticals, 3568 and calcium carbonate, particle shapes of,
Keratinocytes, 1318 Labile polyesters, as biodegradable polymer, 3277
differentiation, 1311 176 consolidation mechanism, 3681
Keratoplasty rejection model Laboratory animals crystallinity of, 4108
in rat, 1196 care of, 124–125 differential scanning calorimetry of,
Ketamine governmental guidelines, 125 2376
in enantiomeric ratio of, 3967 sources of, 124 diluent in tablets, 3655
hydrochloride intramuscular injection site use of, 2830 as filler and flow enhancer, 2081
in cats, 3955 Laboratory autoclaves, bio-validation of, 11 fragmentation, 3681
intravenous injection of, 3967 Laboratory automation, 740 granules
systemic clearances, 3967 Laboratory Information Management System flow of, 3276
Ketobemidone, 1078 (LIMS) particle size of, 3276
bioavailability of, 1078 accreditation and certification standards, Karl Fisher titrations for, 2377
sublingual or buccal route, 1078 2167 mixture of direct compression, 3677
Ketoconazole, 1353 analytical methods, 2164, 2169 monohydrate, 84
3-Keto-desogestrel, 1350 audit trial, 2166 flow properties of, 3291
Ketoprofen, 2408 built in-house systems, 2165, 2169 placebo tablets
delivery, 2127 computerized instrument, 2165 coating, 1234
dispersion, prepared by freeze–drying cost of purchasing, 2166 disintegration rate, 1234
process, 775 definition, 2164 potential residues in, 1615
transdermal absorption of, 2152 detection limits, 2164 powders
in wax, 4070 documentation of, 2164 avalanching behavior of, 3277
Key validation elements, 707 error handling, 2166 flow properties of, 3277
Kidney external standard, 2167 packing characteristics of, 3277
cells baby, 261 generic name, 2164 yield loci for, 3287
function estimation of, 586 implementation, 2165 rotating drum avalanche data of, 3292
glomerulus, 354, 1326 analytical methods, 2169 sorption isotherms for, 2373
tubular epithelium, 156 built-in functionality, 2169 sprayed and non-spray dried, flow
Killion extruder, 2014 organizational issues, 2169 properties of, 3290
Kinematic viscosity, 3130 validation, 2169