12/9/2021
SEAFAST Center, and
Department of
Food Science and Technology
GOOD LABORATORY PRACTICES
Prof. Dr. Lilis Nuraida, MSc.
Outline
• Significance of microbiological analyses results
• Source of Variability and Quality Control in Microbiological
Analyses
• Principles of GLP
• Quality Assurance Program in Microbiological Analyses
• Safety Microbiological Laboratory
Lilis Nuraida – Good Laboratory Practices– December 9, 2021
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Significance of microbiological analyses
results
• The microbiological analysis of food is
part of food safety management and
conformity tests that
define microbiological criteria or assess
the performance of control strategies
based on Food Safety Management
system such as the Hazard Analysis and
Critical Control Point
Lilis Nuraida – Good Laboratory Practices– December 9, 2021
Result of analyses
• Are your data valid??
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Source of variation
• Sampling technique
• Sample storage
• Equipment
• Media/reagents
• Measurement conditions
• Counting effects
• Analyst Effect
• Other random effects
Lilis Nuraida – Good Laboratory Practices– December 9, 2021
Personal Competence
Good Laboratory Practices
Lilis Nuraida – Good Laboratory Practices– December 9, 2021
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Good Laboratory Practice (GLP)
• DEFINITION
• Good Laboratory Practice (GLP)
embodies a set of principles that
provides a framework within which
laboratory studies are planned,
performed, monitored, recorded,
reported and archived.
• GLP helps assure regulatory authorities that
the data submitted are a true reflection of
the results obtained during the study and
can therefore be relied upon when making
risk/safety assessments
Lilis Nuraida – Good Laboratory Practices– December 9, 2021
Objectives of GLP
• The main purpose of the requirements of the
Principles of GLP is having confidence in the
quality, the integrity of the data and being able to
reconstruct activities performed during the
conduct of analyses or research.
• GLP makes sure that the data submitted are a true
reflection of the results that are obtained during
analyses or research.
• GLP also makes sure that data is traceable.
• Promotes international acceptance of tests.
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Standard Operative Procedure (SOP) in
GLP
• Routine inspection, cleaning, maintenance,
testing and calibration.
• Action to be taken in response to
equipment failure.
• Analytical methods.
• Definition of raw data.
• Keeping records, reporting, storage, mixing
and retrieval of data.
Lilis Nuraida – Good Laboratory Practices– December 9, 2021
Area of GLP in Microbiology Lab
• Aseptic Technique
Qualification of reagents and
• Reagents and Media: Preparation,
Storage, Qualification media:
• Reference Standards (Reference How often?
Materials) and Reference Cultures Routinely, (validated
(control of test strains) vs. None validated)
• Equipment and Instruments What to do:
• Method Validation and Verification pH,
• Laboratory Controls Growth promotion,
• Recordkeeping and Documentation: Sterility check
• diligent recording and evaluation of
data
• Employee Training
• Sample Control
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Use of reference standard and cultures
• The use of reference standard/materials and culture is
important for
• Quality control of microbiological test
• Establishing acceptable performance of media
• productivity, selectivity, performance
• Evaluation and interpretation of result.
• Validation of methods
• Verification of test method or kit before use
• To compare between microbiological techniques
• Assessing/evaluating the laboratory performance.
Lilis Nuraida – Good Laboratory Practices– December 9, 2021
Reference materials
• Reference materials (RMs) are indispensable tools in analytical sciences
• RMs are at the edge of building confidence in the comparability of
measurements.
• Widely used for the internal quality control of analytical tests, in the validation of
methodologies as well as verification (accuracy) and as test items in proficiency tests.
• Several RM producers worldwide produce and distribute RMs for
microbiological analysis.
• Prominent RM formats include spray-dried contaminated milk powder (from IRMM, EC;
and RIVM, NL), dried plano-convex discs (from HPA, UK), freeze-dried ‘‘BioBalls’’ (from
BTF, Australia) or dried semi-solid starch matrix materials.
• Proper use of reliable RMs in microbiological analysis is essential to support
classical fields of application, such as method development, method validation
and quality control
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Reference Cultures
• Microbial cultures are delicate standards.
• Procedures should specify careful handling instructions.
• Preparation and resuscitation of cultures should follow the instructions
of the supplier or a validated, established method.
• Using the "Seed-Lot" technique for storage of stock cultures, i.e., using
working cultures and never returning unused passages back to original
stock.
• There should be an established maximum number of passages (5 or less),
and maximum storage time for working cultures.
• Cultures for use in compendial tests should be acquired from a national
culture collection, in frozen, freeze dried, on slants, or in ready-to-use
forms.
• Confirmation of the purity and the identity should be performed
prior to its use in quality control testing.
• Ready-to-use cultures may require additional confirmation of
inoculums size.
Lilis Nuraida – Good Laboratory Practices– December 9, 2021
Validation and Verification
• Validation and verification are important aspects for testing of (food) products
• Validation:
• The confirmation by examination and the provision of objective evidence that the particular
requirements for a specific intended use are fulfilled.
• Confirming that the testing method works to detect the targets in the manner specified
• Verification
• Demonstration that a previously validated method can meet the analytical requirements (i.e.
precision, accuracy, interferences, matrices, analyst) and suitability in the lab/facility.
• Fit for use – an established method will meet customer or laboratory requirements for detection
limits, sample type etc
• To verify that their operators using their equipment in their laboratory environment can apply the
standard method, obtaining the same outcomes as defined in the validation data provided in the
standard method.
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General requirement of Lab’s QA
• A QA program maybe defined as a mechanism used to
ensure that the data are fully reliable, suitable for the
intended purpose, presented on time and at an acceptable
cost.
• Requirements:
• The use of validated methods
• Conduct verification
• The use of standard operating procedure (SOP) in the laboratory
• Calibration and traceability of measurement (including use of certified
reference material)
• External assessment of performance
Lilis Nuraida – Good Laboratory Practices– December 9, 2021
Three essential components QA
programs
1. Prevention, which requires an orderly programs of planning and
positive actions before or during analysis to ensure that all
analytical systems are performing appropriately, easy calibration
and maintenance of instruments, use of reference materials and
training.
2. Assessment, a form of control that includes periodic checks on
the analyst performance e.g. analysis of check samples and
validation of methodology.
3. Correction, an action taken to determine cause(s) of quality
defects and to re-establish proper functioning of analytical
operations e.g. trouble shooting to correct malfunctioning
equipment, re-evaluation of methodology and re-training.
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External Proficiency Testing:
• Proficiency testing is the part of QA programs, which looks at the
accuracy (correctness’) of the results actually being reported by the
laboratory on real test materials.
• An independent external assessment of the correctness of the
typical result provides an impartial test of analytical quality:
• this is done by proficiency testing scheme, i.e. methods of checking
laboratory testing performance by means of inter laboratory tests.
• This includes comparison of a laboratory‘s results at intervals with
those of other laboratories.
• Procedures used when analyzing test materials for such a scheme
are those normally used by the laboratory.
Lilis Nuraida – Good Laboratory Practices– December 9, 2021
Internal Quality Control
• Is designed to check that a laboratory will produce the same result or
outcome if the test or procedure is done on different occasions (within-
laboratory variation), or by different technicians (between-operator
variation).
• is based on between-run precision, closeness of agreement between
results obtained in separate runs of analysis.
• ensures that factors determining the magnitude of uncertainty do not
change during the routine use of an analytical method over long periods
of time.
• is conducted by inserting one or more control materials into every run
of analysis.
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Quality control in microbiological analyses
• Use validated methods
• Use of positive & negative control during test
• Duplicate analyses
• Establishing Reproducibility/Precision
• Spike study
• Participation in Proficiency Test program
• Controlling all aspects affecting test results:
• Sterilities
• Contamination
• Personnel competence
• Facility and equipment
Lilis Nuraida – Good Laboratory Practices– December 9, 2021
Internal Quality Control Checks:
• Blank Analysis:
• Blanks are to included in analytical methods .
• A blank is characterized as a sample included in the analytical processes, which has all the
properties of the actual sample except that it does not contain the substance of interests.
• Duplicate Analysis:
• Duplicate sample analysis is the analysis of the same sample twice in order to determine the
precision of the analysis.
• Spike Analysis:
• A sample is split into two sub samples in the laboratory.
• One is analyzed according to the specified procedure.
• The other is treated by adding a known amount and concentration of the indicator being
measured, running this specified procedures.
• This should increase the concentration in the spiked sample relative to the unspike sample,
by a predictable amount.
• They are used to test the accuracy of the laboratory method.
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External Performance Evaluation/Assessment:
• Evaluation of laboratories based on:
• personnel,
• physical facility,
• instrumentation For this purpose, an
organization should
• quality assurance/quality control programs, and
participate in inter
• the laboratory’s performance.
laboratory comparisons
• To verify that a laboratory possesses the of the proficiency
capability to provide accurate and reliable testing programs
test data in its day to day operations
• To maintain high standards of performance, a
competent, disinterested third party
Lilis Nuraida – Good Laboratory Practices– December 9, 2021
SAMPLING & SAMPLE HANDLING:
• The sampling is a significant role in testing activities as it reflects the
ultimate test results
• The main factors to be considered when taking, and/or storing, samples
for analysis are:
• correct order of sampling – especially if multiple tests are to be conducted
• ensuring that the sample is representative of the lot
• preventing cross-contamination
• preventing degradation, of the sample and/or the measurand
• the reason for the test, e.g. ongoing quality control, compliance with Regulations
or product investigations.
• any statistical analyses that may be required.
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Sample Handling
• The sample should be protected against extraneous contamination.
• The temperature at the time of collection and upon receipt is useful to
the laboratory for the interpretation of results.
• A sample container should not be more than three quarters full in order
to avoid leakage and to allow proper mixing of the sample in the
laboratory.
• Identify samples clearly and completely, and record the sample
information in log book.
Lilis Nuraida – Good Laboratory Practices– December 9, 2021
Safety of microbiological lab
• The microbiology laboratory is a unique
environment that requires special practices and
containment facilities in order to properly protect
persons working with micro-organisms.
• The three main elements of safe containment of
micro-organisms are:
• (1) Good laboratory practices and technique,
• (2) Safety equipment and
• (3) Facility design.
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Biosafety Level of Microbiological lab
Biosafety Examples Agents
Level
1 Bacillus subtilis, Naegleria gruberi, many Escherichia coli, Infectious canine Hepatitis
Virus
2 Bacillus anthracis, Bordetella pertussis, Brucella SPP., Cryptococcus neoformans,
Clostridium botulinum, Clostridium tetani, Helicobacter pylori, most Salmonella spp.,
Yersinia pestis, Mycobacterium leprae, Shigella spp., Human Immunodeficiency Virus,
Human blood.
3 Mycobacterium tuberculosis, Salmonella type, Vesicular stomatitis virus, yellow fever
virus, Francisella tularensis, Coxiella burnetii.
4 smallpox virus, Ebola virus, hemorrhagic fever viruses.
http://www.cdc.gov/OD/ohs/biosfty/bmbl4/bmbl4toc.htm
Lilis Nuraida – Good Laboratory Practices– December 9, 2021
Conclusion
• The general objective of the laboratory might be defined as
producing analytical data of adequate accuracy and reliability
within an acceptable time.
• Good Laboratory Practice (GLP) is to ensure the quality and
integrity of test data related
• Each testing laboratory should developed QA program
• While the reliability of the results has to be fulfill, safety of
microbiological lab is also of primary concern
Lilis Nuraida – Good Laboratory Practices– December 9, 2021
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s1
Thankyou
Prof. Dr. Lilis Nuraida, MSc.
SEAFAST Center and
Department Food Science and
Technology
IPB University
[email protected] Lilis Nuraida – Good Laboratory Practices– December 9, 2021
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