Bioavailability and

Bioequivalence Studies
Dr. Rajan Kalamkar
Associate Professor
Vivekanand Education Society's College of Pharmacy
• The therapeutic effectiveness of a drug depends
upon the ability of the dosage form to deliver the
medicament to its site of action at a rate and
amount sufficient to elicit the desired
pharmacological response.
• This attribute of the dosage form is referred to as
physiological availability, biological availability or
simply, bioavailability.
• For most drugs, the pharmacological response
can be related directly to the plasma levels.
• Thus, the term bioavailability is defined as the
rate and extent (amount) of absorption of
unchanged drug from its dosage form.
 Primary stages of development of a suitable dosage form
for a new drug entity to obtain evidence of its
therapeutic utility.

Determination of influence of excipients, patient related

factors and possible interaction with other drugs on the
efficiency of absorption.

Objectives of
Development of new formulations of the existing drugs.
Bioavailability
Studies
Control of quality of a drug product during the early
stages of marketing in order to determine the influence
of processing factors, storage and stability on drug
absorption.

Comparison of availability of a drug substance from

different dosage forms or from the same dosage form
produced by different manufacturers.
➢ Absolute Bioavailability
Compares the bioavailability of the active drug in systemic
circulation following non-intravenous administration with the same
drug following intravenous administration

✓ For drugs administered intravenously, bioavailability is 100%

✓ Determination of the best administration route

Fab = (AUC)drug
(AUC)IV
➢ Relative Bioavailability

Compares the bioavailability of a formulation (A) of a certain

drug when compared with another formulation (B) of the same
drug, usually an established standard

F rel = ( AUC) drug

(AUC) standard
 AUC: Trapezoidal Rule

AUC2-3 = Cp2 + Cp3 x (t3 - t2)

2
Measurement of Bioavailability
I. Pharmacokinetic Methods
These are very widely used and based on the
assumption that the pharmacokinetic profile
reflects the therapeutic effectiveness of a drug.
Thus, these are indirect methods.
The two major pharmacokinetic methods are:
1. Plasma level-time studies.
2. Urinary excretion studies.
II. Pharmacodynamic Methods
These methods are complementary to
pharmacokinetic approaches and involve direct
measurement of drug effect on a physiological
process as a function of time. The two
pharmacodynamic methods involve determination
of bioavailability from:
1. Acute pharmacological response.
2. Therapeutic response.
Plasma Level—Time Studies
• Unless determination of plasma drug concentration is difficult or impossible,
it is the most reliable method and method of choice in comparison to urine
data.
• The method is based on the assumption that two dosage forms that exhibit
superimposable plasm level time profiles in a group of subjects should result
in identical therapeutic activity.
• With single dose study, the method requires collection of serial blood
samples for a period of 2 to 3 biological half-lives after drug administration,
their analysis for drug concentration and making a plot of concentration
versus corresponding time of sample collection to obtain the plasma level-
time profile.
• With i.v. dose, sampling should start within 5 minutes of drug
administration and subsequent samples taken at 15 minute intervals. To
adequately describe the disposition phase, at least 3 sample points should be
taken if the drug follows one-compartment kinetics.
• For oral dose, at least 3 points should be taken on the ascending part of the
curve for accurate determination of Ka. The points for disposition or
descending phase of the curve must be taken in a manner similar to that for
i.v. dose.
Parameters
1. Cmax: The peak plasma concentration that gives an indication
whether the drug is sufficiently absorbed systemically to provide a
therapeutic response. It is a function of both the rate and extent of
absorption. Cmax will increase with an increase in the dose, as well
as with an increase in the absorption rate.

2. tmax: The peak time that gives an indication of the rate of

absorption. It decreases as the rate of absorption increases.

3. AUC: The area under the plasma level-time curve that gives a
measure of the extent of absorption or the amount of drug that
reaches the systemic circulation.
• The
extent of bioavailability can be determined
by following equations
Urinary Excretion Studies
• This method of assessing bioavailability is based on the
principle that the urinary excretion of unchanged drug is
directly proportional to the plasma concentration of drug.
• As a rule of thumb, determination of bioavailability using
urinary excretion data should be conducted only if at
least 20% of administered dose is excreted unchanged in
the urine. The study is particularly useful for –
Drugs extensively excreted unchanged in the urine – for
example, certain thiazide diuretics and sulphonamides.
Drugs that have urine as the site of action - for example,
urinary antiseptics such as nitrofurantoin and hexamine.
The method involves –
Collection of urine at regular intervals for a time-span equal
to 7 biological half lives.
Analysis of unchanged drug in the collected sample.
Determination of the amount of drug excreted in each
interval and cumulative amount excreted. For obtaining valid
results, following criteria must be met further –
At each sample collection, total emptying of the bladder is
necessary to avoid errors resulting from addition of residual
amount to the next urine sample.
Frequent sampling of urine is also essential in the beginning
in order to compute correctly the rate of absorption.
The fraction excreted unchanged in urine must remain
constant.
The three major parameters examined in urinary excretion
data obtained with a single dose study are:
1. (dXu/dt)max: The maximum urinary excretion rate, it
is obtained from the peak of plot between rate of
excretion versus midpoint time of urine collection period.
It is analogous to the Cmax derived from plasma level
studies since the rate of appearance of drug in the urine is
proportional to its concentration in systemic circulation.
Its value increases as the rate of and/or extent of
absorption increases.
2. (tu)max: The time for maximum excretion rate, it is
analogous to the tmax of plasma level data. Its value
decreases as the absorption rate increases.
3. Xu : The cumulative amount of drug excreted in the
urine, it is related to the AUC of plasma level data and
increases as the extent of absorption increases.
Acute Pharmacological Response
Method
• When bioavailability measurement by
pharmacokinetic methods is difficult, inaccurate or
non-reproducible, an acute pharmacological effect
such as a change in ECG or EEG readings, pupil
diameter, etc. is related to the time course of a given
drug.
• Bioavailability can then be determined by
construction of pharmacological effect-time curve as
well as dose-response graphs.
• The method requires measurement of responses for
at least 3 biological half-lives of the drug in order to
obtain a good estimate of AUC.
Disadvantages–
1. The pharmacological response tends to be more
variable and accurate correlation between
measured response and drug available from the
formulation is difficult.
2. The observed response may be due to an active
metabolite whose concentration is not
proportional to the concentration of parent drug
responsible for the pharmacological effect.
Therapeutic Response Method
Theoretically the most definite, this method is based on observing the
clinical response to a drug formulation given to patients suffering from
disease for which it is intended to be used.
Limitations
1. Quantitation of observed response is too improper to allow for
reasonable assessment of relative bioavailability between two dosage
forms of the same drug.
2. Bioequivalence studies are usually conducted using a crossover
design in which each subject receives each of the test dosage forms,
and it is assumed that the physiological status of the subject does not
change significantly over the duration of the study.
3. Unless multiple-dose protocols are employed, a patient who required
the drug for a disease would be able to receive only a single dose of the
drug every few days or perhaps each week.
4. Many patients receive more than one drug, and the results obtained
from a bioavailability study could be compromised because of a drug–
drug interaction.
BIOEQUIVALENCE
STUDIES
 Need/Objectives for Biequivalence Studies
• During drug development, bioequivalence studies are used to
compare
(a) early and late clinical trial formulations;
(b) formulations used in clinical trials and stability studies, if
different;
(c) clinical trial formulations and to-be-marketed drug products,
if different; and
(d) product strength equivalence, as appropriate.
▪ Bioequivalence study designs are used to support new
formulations of previously approved products, such as a new
fixed-dose combination version of two products approved for
coadministration, or modified-release versions of immediate-
release products.
▪ Postapproval, in vivo bioequivalence studies may be needed to
support regulatory approval of major changes in formulation,
manufacturing, or site, in comparison to reference formulation
(usually the prechange formulation
 Types
Equivalence: It is a relative term that compares drug products with
respect to a specific characteristic or function or to a defined set of
standards. There are several types of equivalences.
Chemical Equivalence: It indicates that two or more drug products
contain the same labelled chemical substance as an active ingredient in
the same amount.
Pharmaceutical Equivalence: This term implies that two or more drug
products are identical in strength, quality, purity, content uniformity and
disintegration and dissolution characteristics; they may however differ in
containing different excipients.
Therapeutic Equivalence: This term indicates that two or more drug
products that contain the same therapeutically active ingredient elicit
identical pharmacological effects and can control the disease to the same
extent.
Bioequivalence: It is a relative term which denotes that the drug
substance in two or more identical dosage forms, reaches the systemic
circulation at the same relative rate and to the same relative extent i.e.
their plasma concentration-time profiles will be identical without
significant statistical differences.
Reference Product
✓ Identified by the Regulatory Authorities as
“Designated Reference Product”

✓ Usually the Global Innovator’s Product

✓Protected by a patent

✓Marketed under manufacturers brand name

✓ Clinical efficacy & safety profile is well documented in

extensive trials

✓ All generics must be Bioequivalent to it

✓ In India, CDSCO may approve another product as Reference product

Generic Drug
✓Drug product which is identical or bioequivalent to Brand/ Reference
drug in:
• Active ingredient (s)
• Route of administration
• Dosage form
• Strength
• Indications
• Safety

✓May have different:

• Inactive ingredients
• Colour
• Shape

✓ Almost half of drugs in market have Generics

Price difference between
Reference & Generic Drugs

Reference Drug Generic Drug

• Expensive • 30-80% cheaper
• 5/5000 new drug candidates • Since already tested & approved,
tested in humans & 1 approved cost of simply manufacturing

• Takes 12-15 yrs • Fraction of the cost of testing &

development
• Costs around 1 billion $
• Drug Patents of 20yrs, applied • Approved for sale after drug
before clinical trials begin patent protection expires

• Effectively 7-12 yrs

Study Design
✓Good experimental design, enhances the power of the study

✓ Depends on: question to be answered, nature of reference

drug/ dosage form, benefit-risk ratio

✓ As far as possible, the study should be of crossover design &

suitably randomized

✓Ideal design: Randomized two-period, two-sequence,

Crossover design with adequate washout period

✓If the half-life is long: Parallel design

✓For highly variable drugs: Replicate design

Types of Bioequivalence Studies
Bioequivalence can be demonstrated either –
▪ In vivo, or
▪ In vitro.
Bioequivalence Experimental Study Design
1. Completely randomised designs
• In a completely randomised design, all treatments
(factor levels) are randomly allocated among all
experimental subjects.
Method of randomisation
• Label all subjects with the same number of digits,
for e.g., if there are 20 subjects, number them from
1 to 20. Randomly select non-repeating random
numbers (like simple randomisation) with among
these labels for the first treatment, and then repeat
for all other treatments.
Advantages
1) The design is extremely easy to construct.
2) It can accommodate any number of treatments and subjects.
3) The design is easy and simple to analyse even though the
sample sizes might not be the same for each treatment.
Disadvantages
1) Although the design can be used for any number of
treatments, it is best suited for situations in which there are
relatively few treatments.
2) All subjects must be as homogeneous as possible. Any
extraneous sources of variability will tend to inflate the
random error term, making it difficult to detect differences
among the treatment (or factor level) mean responses.
2. Randomised block designs
First, subjects are sorted into homogeneous groups,
called blocks and the treatments are then assigned at
random within the blocks.
Method of Randomisation
Subjects having similar background characteristics
are formed as blocks. Then treatments are randomised
within each block, just like the simple randomisation.
Randomisations for different blocks are done
independent of each other.
• Advantages
1) With effective and systematic way of grouping, it can provide
substantially more precise results than a completely randomised design
of comparable size.
2) It can accommodate any number of treatments or replications.
3) Different treatments need not have equal sample size.
4) The statistical analysis is relatively simple. The design is easy to
construct.
5) If an entire treatment or block needs to be dropped from the analysis
for some reason, such as spoiled results, the analysis is not thereby
complicated.
6) Variability in experimental units can be deliberately introduced to
widen the range of validity of the experimental results without
sacrificing the precision of results.
• Disadvantages
1) Missing observations within a block require more complex analysis.
2) The degrees of freedom of experimental error are not as large as with
a completely randomised design.
3. Repeated measures, cross-over and carry-
over designs
• This is essentially a randomised block design in which the same subject
serves as a block.
• The same subject is utilized for each of the treatments under study. Since
we take repeated measures on each subject we get the design name
―repeated measures design‖.
• The study may involve several treatments or a single treatment evaluated at
different points in time.
• The administration of two or more treatments one after the other in a
specified or random order to the same group of patients is called a
crossover design or change-over design.
• The drawback of crossover studies is the potential for distortion due to
carry-over, that is, residual effects from preceding treatments.
• To prevent carry-over effects, one must always allow for a wash-out period
during which most of the drug is eliminated from the body – generally
about 10 elimination half-lives.
Example: clinical trials to monitor safety and side effects.
 ✓2 formulations, even number of subjects, randomly divided into 2
equal groups
✓First period , each member of one group receive a single dose of the
test formulation; each member of the other group receive the standard
formulation
✓After a wash period (5 half lives), in second period , each member of
the respective groups will receive an alternative formulation &
experiment will be repeated.
Subjects Period 1 Period 2

1-8 T S

9-16 S T
Method of Randomisation
• Complete randomisation is used to randomise the order
of treatments for each subject.
• Randomisations for different subjects are independent of
each other.
Advantages
1) They provide good precision for comparing treatments
because all sources of variability between subjects are excluded
from the experimental error.
2) It is economic on subjects. This is particularly important
when only a few subjects can be utilized for the experiments.
3) When the interest is in the effects of a treatment over time, it
is usually desirable to observe the same subject at different
points in time rather than observing different subjects at the
specified points in time.
Disadvantages
1) There may be an order effect, which is connected with the
position in the treatment order.
2) There may be a carry-over effect, which is connected with
the preceding treatment or treatments.
4. Latin square designs
• Completely randomised design, randomised block design and repeated measures
design are experiments where the person/subject/volunteer remains on the
treatment from the start of the experiment until the end and thus are called as
continuous trial.
• In a Latin square, however, each subject receives each treatment during the course
of the experiment.
• A Latin square design is a two-factor design (subjects and treatments are the two
factors) with one observation in each cell.
• Such a design is useful compared the earlier ones when three or more treatments
are to be compared and carry-over effects are balanced.
• In a Latin square design, rows represent subjects, and columns represent
treatments.
• A r x r Latin square design is a square with r rows and r columns such that each
of the r2 cells contain one and only one of the r letters representing the
treatments, and each letter appears once and only once in ever row and every
column.
• A Latin square is called standard if the first row and the first column consist of
the r letters in alphabetical order.
• Randomised, balanced, cross-over Latin square design are commonly used for
bioequivalence studies.
Latin Square Design
✓ More than two formulations

✓ A group of volunteers will receive formulations in the sequence

shown
Advantages
1) It minimizes the inter-subject variability in plasma drug
levels.
2) Minimizes the carry-over effects which could occur when a
given dosage form influences the bioavailability of a
subsequently administered product (intrasubject variability).
3) Minimizes the variations due to time effect.
4) Treatment effects can be studied from a small-scale
experiment. This is particularly helpful in preliminary or pilot
studies.
5) Makes it possible to focus more on the formulation
variables which is the key to success for any bioequivalence
study.
Disadvantages
1) The use of Latin square design will lead to a very small
number of degrees of freedom for experimental error when
only a few treatments are studied. On the other hand, when
many treatments are studied, the degrees of freedom for
experimental error maybe larger than necessary.
2) The randomisation required is somewhat more complex than
that for earlier designs considered.
3) The study takes a long time since an appropriate washout
period between two administrations is essential which may be
very long if the drug has a long t½.
4) When the number of formulations to be tested is more, the
study becomes more difficult and subject dropout rates are also
high. This can be overcome by use of a balanced incomplete
block design in which a subject receives no more than 2
formulations.
IV. Parallel-Group Design
✓Even number of subjects in two groups

✓Each receive a different formulation

✓No washout necessary

✓For drugs with long half life

Treatment A Treatment B
1 2

3 4

5 6

7 8

9 10

11 12
 Parallel Crossover

• Groups assigned different • Each patient receives both

treatments treatments

• Shorter duration • Longer duration

• Larger sample size • Smaller sample size

• No carryover effect • Carryover effect

• Doesn’t require stable disease • Requires stable disease & similar

& similar baseline baseline
Subject selection

✓Healthy adult volunteers

✓Age: 18-45 yrs

✓Age/Sex representation corresponding to therapeutic & safety profile

✓Weight within normal limits→ BMI

✓Women: Pregnancy test prior to 1st & last dose of study; OC pills C/I

✓Drug use intended in Elders (Age >60yrs)

✓Teratogenic Drugs→ Male volunteers

✓Highly toxic drugs: Patients with concerned disease (stable) eg.

Cancer
Exclusion Criteria
✓H/o allergy to test drug
✓H/o liver or kidney dysfunction
✓H/o jaundice in past 6 months
✓Chronic diseases eg. Asthma, arthritis
✓Psychiatric illness
✓Chronic smoker, alcohol addiction, drug abuse
✓Intake of enzyme modifying drug in past 3 months
✓Intake of OTC/Prescription drugs past 2 weeks
✓HIV positive
✓BA & BE studies in past 3 months
✓H/o bleeding disorder
Selection of Number of Subjects
✓ Sample size is estimated by:
• Pilot experiment
• Previous studies
• Published data

✓ Significance level desired, usually 0.05

✓ Power of the study, normally 80% or more

✓ Expected deviation (Δ) from the reference product,

as compatible with BE

✓ If no data available, reference ratio of 0.95 (Δ = 5%) used

✓ Minimum 16 subjects, unless ethical justification

✓ Allow for drop-outs

✓ Replace drop-outs→ substitute follow same protocol;

similar environment

✓ Sequential/ Add-on Studies→ large no. of subjects required,

results of study do not convey adequate significance
Characteristics to be measured
✓ Accessible biological fluids like blood, plasma &/or serum
to indicate release of the drug substance from the drug
product into the systemic circulation

✓Mostly: Active drug substance

✓Active / Inactive metabolite maybe measured in cases of:

• Concentration of drug too low

• Limitation of analytical method
• Unstable drug
• Drug with very short half life
• Pro-drugs

✓ Excretion of drug & its metabolites in urine→ Non-linear kinetics

✓ Measure individual enantiomers when they exhibit:

• Different pharmacokinetic/ pharmacodynamic properties

• Non-linear absorption
• Safety/Efficacy purposes

✓ Drugs that are not absorbed systemically from site of application

surrogate marker needed for BA & BE determination
Blood Sampling points/ Schedule
Single-dose study of an immediate release product:

✓ For at least three elimination half-lives (cover >80% of AUC)

• Absorption phase : 3-4 points
• Around Tmax : 3-4 points
• During elimination : 4 points
✓ Intervals not longer than the half-life of the drug
✓ If urine tested, collect it for at least 7 half-lives
Method Validation
✓ Accuracy/ Relative Recovery
Closeness of determined value to the true Value

✓ Precision
Closeness of agreement obtained from the multiple sampling of the
same homogeneous samples under certain prescribed conditions

• Repeatability
Precision under same conditions
same analyst, same apparatus, same interval of time, identical
reagents

• Reproducibility
Precision under different conditions
different analysts, apparatus from different manufacturers,
different days, reagents from different sources
✓ Sensitivity
Capacity of the test procedure to record small variations in
concentration

• Limit of detection (LOD):

Lowest concentration of drug that will yield an assay response
significantly different from that of a sample blank

• Limit of quantitation (LOQ/ sensitivity limit):

Lowest concentration of drug that can be determined with acceptable
precision & accuracy under the stated experimental conditions

✓ Selectivity/ Specificity
Ability of the method to measure only what it is intended to measure
✓ Calibration of Instruments

• Should be done regularly & as per standard procedures in

USP/BP/IP

• Done before starting the analysis at the development phase &

during the study phase

✓ Predetermined SOPs

✓ Accreditation of analyzing laboratory

Parameters to be measured
✓Pharmacokinetic Parameters measured are:

• Cmax
• Tmax
• AUC0-t
• AUC0-∞ AUC 0-∞ = AUC 0-t + Clast /k
For steady state studies:

• AUC0-t
• Cmax
• Cmin
• Degree of fluctuation
Fasting & Fed State Conditions

➢ Fasting Conditions:
✓ Single dose study:

• Overnight fast (10 hrs) and subsequent fast of 4 hrs

✓ Multiple dose study:

• Two hours fasting before and after the dose
➢ Fed State Studies
✓Required when:

• Drug recommended with food

• Modified release product
• Assessment of Cmax and Tmax difficult with fasting state study

✓ Requires consumption of a high fat food, 15 minutes before dosing

✓ Provide 950-1000 kcals

✓ Fat- 50%, Proteins 15-20%, Carbohydrate- 30-35%

✓ Ethnic & cultural variation considered

✓ Specified in protocol
 Statistical Evaluation
✓ Primary concern of bioequivalence is to limit Consumer’s &
Manufacturer’s risk

• Cmax & AUC analysed using ANOVA

• Tmax analysed by non-parametric methods

✓ Use natural log transformation of Cmax and AUC

• Calculate Geometric means of Cmax of Test [Cmax’t]

• Calculate Geometric means of Cmax of Reference [Cmax’r]
• Calculate Geometric Mean Ratio= [Cmax’t] / [Cmax’r]

✓ Calculate 90% confidence interval for this GMR for Cmax

✓ Similarly calculate GMR for AUC

To establish BE:

✓ The calculated 90% CI for Cmax & AUC, should fall within range:
80-125% (Range of Bioequivalence)

✓ Non-parametric data 90% CI for Tmax should lie within

clinical acceptable range
Conduct of Study
➢ Pre-study Requirements
✓ IEC approved protocol

✓ Written procedure (SOPs) for all the study related activities

✓ In accordance with ICH-GCP Guidelines

✓ Adequate infrastructure- Clinical facility

✓ Trained Study personnel

✓Healthy Volunteers
➢ Screening of Healthy volunteers

✓ Recruitment through advertisements

✓ Written consent for Screening & Consent for HIV testing
✓ Height & weight
✓ Medical History
✓ Physical examination, ECG & vital signs examination
✓ Blood & Urine sample
(Lab testing,; tests for HIV, Hepatitis A, B & C; UPT→ females)
➢ Volunteer Selection & Recruitment
✓ Volunteers called 1 day before study & admitted
✓ Written ICF taken

➢ During the Study

✓ Standardized study environment
✓ Vital signs examination at scheduled times
✓ Standardised amount of water [~240ml]
✓ No concomitant medications [including herbal remedies]
✓ Administration of the study medication is supervised by the
investigator
✓ Same time of dosing (multiple dosing)
✓ Sampling time with deviation of 2 mins allowed
✓ Uniform & identical meals at identical times in all periods
✓ Restriction of xanthines, grapefruit, citrus fruits, smoking, alcohol
✓ Physical activity & posture standardized→
limit effects on GI flow & motility
✓All activities recorded in CRFs with time & date
➢ End of Study

✓ Post-study examination for safety assessment

✓ Compensation to subjects as per agreed terms
✓ Clinical part of study completed
Documentation
• Signed detailed protocol
• Approval by Ethics Committee
• Volunteer Information sheet
• Informed Consent Form (ICF)
• Case Record Form (CRF)
• Undertaking by investigator
• CV of investigator
• Randomization chart
• Laboratory certification
• Analytical method validation details
• Chromatograms of all volunteers including any aberrant ones
• Tabulated Raw Data of volunteers
Maintenance of Records & Retention of
Study Samples

✓ All Records of in vivo tests on any marketed batch of a

drug product should be maintained by the Sponsor
for atleast 2 years after expiry date of the batch

✓ All Drug samples to be retained for a period of atleast

3 years after conduct of the study

OR

1year after expiry of the batch

[Stored in conditions consistent with the product labeling]
BIOAVAILABILITY ENHANCEMENT
THROUGH ENHANCEMENT OF
DRUG SOLUBILITY OR
DISSOLUTION RATE
Micronization
• The process involves reducing the size of the
solid drug particles to 1 to 10 microns commonly
by spray drying or by use of air attrition methods
(fluid energy or jet mill).
• The process is also called as micro-milling.
• Examples of drugs whose bioavailability have
been increased by micronization include
griseofulvin and several steroidal and sulpha
drugs.
 Nanonisation:
• It‘s a process whereby the drug powder is converted to
nanocrystals of sizes 200 - 600 nm, e.g. amphotericin B. The
main production technologies currently in use to produce drug
nanocrystals yield as a product a dispersion of drug nanocrystals
in a liquid, typically water (called nanosuspension).
• Thereare three basic technologies currently in use to prepare
nanoparticles:
i. Pearl milling
ii. Homogenisation in water (wet milling as in a colloid mill)
iii. Homogenisation in non-aqueous media or in water with
water-miscible liquids.
Use of Surfactants:
• Surfactants are very useful as absorption enhancers and
enhance both dissolution rate as well as permeability of drug.
• They enhance dissolution rate primarily by promoting wetting
and penetration of dissolution fluid into the solid drug
particles.
• They are generally used in concentration below their critical
micelle concentration (CMC) values since above CMC, the
drug entrapped in the micelle structure fails to partition in the
dissolution fluid.
• Nonionic surfactants like polysorbates are widely used.
• Examples of drugs whose bioavailability have been increased
by use of surfactants in the formulation include steroids like
spironolactone.
 Use of Salt Forms:
• Saltshave improved solubility and dissolution characteristics in comparison
to the original drug. It is generally accepted that a minimum difference of 3
units between the pKa value of the group and that of its counterion is required
to form stable salts.
• Alkalimetal salts of acidic drugs like penicillins and strong acid salts of basic
drugs like atropine are more water-soluble than the parent drug. Factors that
influence salt selection are physical and chemical properties of the salt, safety
of counterion, therapeutic indications and route of administration.
Salt formation does have its limitations –
• It is not feasible to form salts of neutral compounds.
• It may be difficult to form salts of very weak bases or acids.
• Thesalt may be hygroscopic, exhibit polymorphism or has poor processing
characteristics.
• Conversionof salt to free acid or base form of the drug on surface of solid
dosage form that prevents or retards drug release.
• Precipitation of unionised drug in the GI milieu that has poor solubility.
Use of Precipitation Inhibitors:
•A significant increase in free drug concentration above
equilibrium solubility results in supersaturation, which can
lead to drug precipitation or crystallization.
• This can be prevented by use of inert polymers such
HPMC, PVP, PVA, PEG, etc. which act by one or more of
the following mechanisms -
➢ Increase the viscosity of crystallization medium thereby
reducing the crystallization rate of drugs.
➢ Provide a steric barrier to drug molecules and inhibit
crystallization through specific intermolecular interactions
on growing crystal surfaces.
➢ Adsorb onto faces of host crystals, reduce the crystal
growth rate of the host and produce smaller crystals.
Alteration of pH of the Drug Microenvironment:
This can be achieved in two ways—in situ salt formation, and
addition of buffers to the formulation e.g. buffered aspirin
tablets.

Use of Amorphs, Anhydrates, Solvates and Metastable

Polymorphs:
Depending upon the internal structure of the solid drug,
selection of proper form of drug with greater solubility is
important. In general, amorphs are more soluble than
metastable polymorphs, anhydrates are more soluble than
hydrates and solvates are more soluble than non-solvates.
Selective Adsorption on Insoluble Carriers:

•A highly active adsorbent such as the inorganic clays

like bentonite can enhance the dissolution rate of
poorly water-soluble drugs such as griseofulvin,
indomethacin and prednisone by maintaining the
concentration gradient at its maximum.
• The two reasons suggested for the rapid release of
drugs from the surface of clays are—the weak
physical bonding between the adsorbate and the
adsorbent, and hydration and swelling of the clay in
the aqueous media.
Solid Solutions:
• The three means by which the particle size of a drug can be
reduced to submicron level are—
▪ Use of solid solutions,
▪ Use of eutectic mixtures, and
▪ Use of solid dispersions.
• In all these cases, the solute is frequently a poorly water-soluble
drug acting as the guest and the solvent is a highly water-
soluble compound or polymer acting as a hos or carrier.
• A solid solution is a binary system comprising of a solid solute
molecularly dispersed in a solid solvent. Since the two
components crystallize together in a homogeneous one phase
system, solid solutions are also called as molecular dispersions
or mixed crystals.
• Because of reduction in particle size to the molecular level,
solid solutions show greater aqueous solubility and faster
dissolution than eutectics and solid dispersions.
Eutectic Mixtures:
• These systems are also prepared by fusion method.
Eutectic melts differ from solid solutions in that the
fused melt of solute-solvent show complete miscibility
but negligible solid-solid solubility i.e. such systems
are basically intimately blended physical mixture of
two crystalline components.
• When the eutectic mixture is exposed to water, the
soluble carrier dissolves leaving the drug in a
microcrystalline state which solubilises rapidly.
Solid Dispersions:
• These are generally prepared by solvent or co-precipitation method
whereby both the guest solute and the solid carrier solvent are
dissolved in a common volatile liquid solvent such as alcohol. The
liquid solvent is removed by evaporation under reduced pressure or
by freeze-drying which results in amorphous precipitation of guest
in a crystalline carrier.
• Thus, the basic difference between solid dispersions and solid
solutions/eutectics is that the drug is precipitated out in an
amorphous form in the former as opposed to crystalline form in the
latter; e.g. amorphous sulphathiazole in crystalline urea. Such
dispersions are often called as coevaporates or co-precipitates.
• The method is suitable for thermolabile substances but has a number
of disadvantages like higher cost of processing, use of large
quantities of solvent, difficulty in complete removal of solvent, etc.
• The carriers used are same as for eutectics or solid solutions. With
glassy materials, the dispersions formed are called as glass
dispersions or glass suspensions.
• Other polymers such as PEG and HPMC are also employed to
prepare solid dispersions of poorly water-soluble drugs such as
nifedipine and itraconazole.
Molecular Encapsulation with Cyclodextrins:
• The beta- and gamma cyclodextrins and several of their
derivatives are unique in having the ability to form
molecular inclusion complexes with hydrophobic drugs
having poor aqueous solubility.
• These bucket-shaped oligosaccharides produced from
starch are versatile in having a hydrophobic cavity of size
suitable enough to accommodate the lipophilic drugs as
guests; the outside of the host molecule is relatively
hydrophilic.
• Thus, the molecularly encapsulated drug has greatly
improved aqueous solubility and dissolution rate. There
are several examples of drugs with improved
bioavailability due to such a phenomenon — thiazide
diuretics, barbiturates, benzodiazepines and a number of
NSAIDs.
BIOAVAILABILITY ENHANCEMENT THROUGH ENHANCEMENT
OF DRUG STABILITY
1. Enteric Coating:

• Enteric-coated systems utilize polymeric

coatings that are insoluble in the gastric media
and therefore, prevent or retard drug release in
the stomach.
• Such systems release the drug in the alkaline
milieu of intestine.
• Bioavailability of drugs that are unstable in the
gastric milieu, for e.g. erythromycin, penicillin V,
pancreatin and benzimidizoles such as
omeprazole can be improved by enteric coating.
2. Complexation:
• Complexation, in certain instances, can be used
to increase the stability of drug in the GI milieu,
particularly those of ester drugs and thus enhance
their oral availability.
• Generally speaking, β-cyclodextrins are potential
carriers for achieving such objectives but other
complexing agents, such as caffeine, sodium
salicylate, sodium benzoate, and nicotinamide,
may also be used.
3. Use of Metabolism Inhibitors:
• Co-administration of a drug with low
bioavailability and its metabolism inhibitor, which
can selectively inhibit any of the contributing
processes, would result in increased fractional
absorption and hence a higher bioavailability.
• In fact, this approach seems to be a promising
alternative to overcome the enzymatic barriers to
oral delivery of metabolically labile drugs such as
peptides and proteins.
Current novel approaches in this area include:
• ➢ Bioadhesive delivery systems that can reduce the
drug degradation between the delivery system and
absorbing membrane by providing intimate contact with
GI mucosa.
• ➢ Controlled-release microencapsulated systems that
can provide simultaneous delivery of a drug and its
specific enzyme inhibitor at the desired site for required
period of time.
•➢ Immobilization of enzyme inhibitors on
mucoadhesive delivery systems.