Bioavailability

&
Bioequivalence

KH. HUSSAN REZA,

MAKAUT,W.B

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 Definitions
 Objectives of Bioavailability studies
 Methods of Bioavailability measurement
--Pharmacokinetic methods:
1. Plasma level time studies
2. Urinary excretion studies
--Pharmacodynamic methods:
1. Acute pharmacological response
2. Therapeutic response
 In vitro dissolution studies and bioavailability
 IVIVC Correlation
 Bioequivalence experimental study designs
1. Completely randomized designs
2. Randomized block designs
3. Repeated measures, cross over, carry-over
designs
4. Latin square designs
 Statistical interpretation of bioequivalence data
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[Link] of variance (ANOVA)

 Definitions
Bioavailability:
It is rate and extent of absorption of unchanged drug
from its dosage form.
Rate- acute conditions- asthma, pain etc
Extent( amount ) – chronic conditions- hypertension.
Influence of route of administration
PARENTRAL> ORAL> RECTAL>TOPICAL

Absolute bioavailability:
When systemic availability of a drug administered
orally
is determined in comparison to its I.V. administration,
denoted by F.

Relative bioavailability:
When systemic availability of a drug after oral
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administration is
Chemical equivalence:
When two or more drug products contain the same
chemical substance as an active ingredient in the same
amount it is called chemical equivalence.

Bioequivalence:
It is relative term that denotes drug substance in two or
more identical dosage forms reaches the systemic
circulation at the same relative rate to the same
relative extent.

i.e. plasma concentration-time profiles will be identical

without significant statistical differences.

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 Objectives of Bioavailability studies

It is important in the
Primary stages of development of dosage form of
new drug entity to find its therapeutic utility.

Determination of influence of excipients on

absorption.

Development of new formulations of existing drugs.

 Control of quality of drug products and influence of

processing factors , storage and stability on

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absorption.
Single dose versus multiple dose studies:
Single dose bioavailability studies are very common, easy, less
exposure to drugs,
less tedious. However it is difficult to predict the steady state
characteristics
and inter subject variability with these studies.
Advantages of multiple dose study:
-Accurately reflects manner in which drug will be used
clinically.
-Requires collection of few blood samples.
-Drug levels are higher due to cumulative effect and
useful for less sensitive analytical methods
-Better evaluation of controlled released formulation.
-Nonlinearity if present , can be easily determined.
-No need of long wash out periods.

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Healthy subjects versus Patients:
Generally bioavailability study should be carried out in patients, as
patient get
benefited from the study, reflects better therapeutic efficacy, drug
absorption
pattern in disease state can be studied, avoids ethical quandary of
administering
drug to healthy subjects. but there are also various drawbacks like
diseases, other
drugs, physiological changes, fasting state is difficult etc. Hence
usually these
studies performed on young (20-40yrs) healthy male adult
volunteers( body weight
±10%) under restricted dietary and fixed activity conditions.

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 2. Urinary excretion studies:
This method is based on the principle that the urinary excretion of
unchanged drug is directly proportional to the plasma concentration
of drug.
It can be performed if
-At least 20% of administered dose is excreted unchanged in urine.
The study is useful for
- Drugs that extensively excreted unchanged in urine eg. Thiazide diuetics
- Drugs that have urine as site of action eg. Urinary antiseptics like
nitrofurontoin.
Steps involved:
-collection of urine at regular intervals for 7 half lifes.
- Analysis of unchanged drug in collected sample.
- Determination of amount of drug at each interval and cumulative as
well.
Criteria's must be followed
- At each sample collection total emptying of bladder is necessary.
- Frequent sampling is essential in the beginning to compute correct
rate of absorption.
- The fraction excreted unchanged in urine must remain constant.
Parameters considered important in Urinary excretion studies9
1. (Dx/dt)max: Maximun urinary excretion rate
2. (tu)max: Time for maximum excretion rate
 Pharmacodynamic methods
1. Acute pharmacological response:
When bioavailability measurement by pharmacokinetic methods is
difficult, inaccurate or non reproducible this method is used. Such as
ECG, EEG, Pupil diameter etc.
It can be determined by dose response graphs. Responses measure for
at least 3 half lifes.
Disadvantages:
- Pharmacological response is variable and accurate correlation drug
and formulation is difficult.
-Observed response may be due to active metabolite.
2. Therapeutic response:
This method is based on observing clinical response in patients.
Drawbacks:
- Quantitation of observed response is too improper.
-The physiological status of subject assumed that does not change
significantly over duration of study.
-If multiple dose protocols are not involved. Patient receive only single
dose for few days or a week 10
-The patient s receiving more than one drug treatment may be
compromised due to drug-drug interaction.
 In vitro dissolution studies and bioavailability:

The physicochemical property of most drugs that has greatest influence on

absorption from GIT is dissolution rate. However in vitro dissolution is good
substitute for in vivo study in terms of saving cost and time. The best
available tool today which can at least quantitatively assure about the
bioavailability of drug from its formulation is in vitro dissolution test.
In vitro- in vivo correlation ( IVIVC):
It is defined as the predictive mathematical model that describes the
relationship between in vitro property ( rate & extent of dissolution) and in
vivo response ( plasma drug concentration).
The main objective of developing and evaluating IVIVC is to use dissolution
test to serve as alternate for in vivo study in human beings.
IVIVC Levels:
Level A: The highest category of correlation. It represents point to point
correlation between in vitro dissolution and in vivo rate of absorption.
Advantages: serves as alternate for in vivo study, change in manf.
Procedure or
formula can be justified without human studies.
Level B: The mean in vitro dissolution time is compare with mean in vivo
residence time. It is not point to point correlation . Data can
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be used for
quality control standards.
Level C: It is single point correlation. e.g. t50%, Tmax, Cmax. This level is
 Types of bioequivalence studies
In vivo bioequivalence studies: when needed,
1. Oral immediate release product with systemic action
-Indicated for serious conditions requiring assured response.
-Narrow therapeutic window.
- complicated pharmacokinetic, absorption <70%, presystemic
elimination>70%, nonlinear kinetics.
2. Non-oral immediate release products
3. Modified release products with systemic action.
In vitro bioequivalence studies: If none of the above criteria is
applicable comparative in vitro dissolution studies can be done .
Biowaivers: In vivo studies can be exempted under certain conditions.
1. Drug product only differ in strength of drug provided,
- Their pharmacokinetics are linear, Drug & excipient ratio is same,
- both products manufactured by same manuf. at same site.
- BA/BE study done for original product, disso. rate same under same
conditions.
2. The method of production slightly modified in a way that not
affect bioavailability
3. The drug product meet following requirements: The product is in
solubilised form, 12

no excipients affecting absorption, Topical use, Oral but not absorbed,

inhalation as gas or vapour.
 Bioequivalence experimental study designs
[Link] randomized designs:
All treatments are randomly allocated among all experimental subjects.
e.g. If there are 20 subjects, number the from 1 to 20. randomly select non
repeating numbers among these labels for the first treatment. And then
repeat for all other treatments .
Advantages: Easy to construct, can accommodate any number of
treatment and subjects, Simple to analyze.
Disadvantages:
Although can be used for number of treatments, but suited for few
treatments.
All subjects must be homogenous or random error will occur.
[Link] block designs:
First subjects are sorted in homogenous groups, called blocks and then
treatments are assigned at random within blocks.
Advantages:
Systematic grouping gives more precise results.
No need o equal sample size, any number of treatments can be followed,
statistical analysis is simple, block can be dropped , variability can be
introduced.
Disadvantages:
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Missing observations in a block require more complex analysis.
Degree f freedom is less.
[Link] measures, cross over designs:
It is a kind of randomized block design where same subject serves as a
block.
Same subject utilized repeatedly so called as repeated measure design.
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 cross-over designs.
Advantages:
Good precision, Economic, can be performed with few subjects, useful in
observing
effects of treatment over time in the same subject.
Disadvantages:
Order effect due to position in treatment order.
Cary over effect due to preceding treatment.
Wash out period necessary – 10 elimination half lifes.
[Link] square designs: All other above designs are continuous trial.
However in
Latin square design each subject receives each treatment during the
experiment.
It is a two factor design ( Rows=Subjects and Columns=Treatments ).
Carry –over
effects are balanced.
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Advantages: minimize variability of plasma profiles and carry-over
effects. Small
 Statistical interpretation of bioequivalence data
After the data has been collected , statistical methods must be applied to
determine
the level of significance or any observed difference in rate and /or extent of
absorption
to establish bioequivalence between two or more drug products.
1. Analysis of varience ( ANOVA): It is statistical procedure use to test
data for differences within and between treatment and control groups. A
statistical difference between the pharmacokinetic parameters obtained
from two or more drug products is considered statistically significant if
there is probability of less than 1 in 20 or 0.05 (p≤0.05) . The value of
p indicates the level of statistical significance.
2. Confidence interval approach: It is also called as two one-sided
procedure and used to demonstrate if bioavailability of test product is too
low or too high in comparison to reference product. 90% confidence
interval of two drug products must be within ±20% for bioavailability
parameters such as AUC or Cmax. ( i.e. between 80 to15102 %). For log
transformed data 90% confidence interval is set at 80-125%.
 References
 Biopharmaceutics and pharmacokinetics – A
Treatise ,
D. M. Brahmankar, Sunil [Link]. Vallabh
prakashan IInd
edition, pp- 315-366.
 Basics of Pharmaokinetics, Leon Shargel,
fifth edition, willey
publications, pp- 453-490.
 Internet sources.

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