BP604T

Unit-II
Bioavailability and
Bioequivalence

Dr Aswathi R Hegde
1
Faculty of Pharmacy © Ramaiah University of Applied Sciences
 Session objectives
By the end of this session, students will be able to:
• Describe the concept and characteristics of bioavailability

• Appreciate the importance of bioavailability studies

• Describe the different methods for the assessment of bioavailability

• Discuss on the various considerations in bioavailability

• Justify the role of bioavailability studies in drug approval process

• Identification of right plan for conducting bioequivalence studies

• Analyze the data produced from the bioequivalence studies

2
Faculty of Pharmacy © Ramaiah University of Applied Sciences
 Measurement of Bioavailability

Plasma level time

studies
Pharmacokinetic methods
(Indirect) Urinary excretion
studies

Acute pharmacological
Pharmacodynamic methods response
(Direct)
Therapeutic response

Clinical observation
3
Faculty of Pharmacy © Ramaiah University of Applied Sciences
 Plasma level time studies
• Most reliable method compared to urine data
• Assumption – two dosage forms that exhibit similar plasma level-time profiles in a group
of subjects should result in identical therapeutic activity
• Plasma level time studies or the plasma concentration – time curve or blood level curve.
• A direct relationship exists concentration of drug at the site of action & concentration of
drug in the plasma.
• Serial blood samples are taken after drug administration & analyzed for drug
concentration.
– For single dose study, collection of blood samples for 2-3 biological half lives
– For i.v. dose – sampling should start within 5 min of drug administration and subsequent samples at
intervals of 15 min
– To describe disposition phase – at least 3 sample points must be taken for one-compartment model
and 5-6 points for two-compartment model
– For oral dose, at least 3 points must be taken on ascending part of curve to determine absorption rate
constant Ka 4
Faculty of Pharmacy © Ramaiah University of Applied Sciences
 Plasma level time studies
Three parameters of plasma level-time studies considered important for determining
bioavailability are:
- Cmax maximum plasma drug concentration obtained after oral administration,
provides an indication whether drug is sufficiently absorbed systematically to provide
therapeutic response; increases with increase in dose and absorption rate
- tmax time required to reach maximum drug conc.; gives indication of rate of
absorption; decreases as rate of absorption increase
- AUC corresponds to measurement of the extent of drug bioavailability; reflects
total amount of drug that reaches the system circulation; gives indication of extent of
absorption.

C max t max

AUC
5
Faculty of Pharmacy © Ramaiah University of Applied Sciences
 6
Faculty of Pharmacy © Ramaiah University of Applied Sciences
 • Extent of Bioavailability determined by

[AUC]oral Div
F = F= Absolute bioavailability
[AUC]iv Doral

[AUC]test Dstd Fr= Relative bioavailability

Fr =
[AUC]std Dtest
With multiple dose study, extent of bioavailability is given by:-
[AUC]test Dstd test
F =
[AUC]std Dtest
Std

7
Faculty of Pharmacy © Ramaiah University of Applied Sciences
 Urinary excretion studies
• Indirect method for estimating BA

• Based on principle that the urinary excretion of unchanged drug is directly

proportional to plasma concentration of drug

• Urine samples must be collected and total amount of drug excreted must be obtained

• At each sample collection, total emptying of the bladder is necessary to avoid error
resulting from addition of residual amounts to the next urine sample

• Used for drugs extensively excreted unchanged in the urine e.g. thiazide diuretics and
drugs that have urine as site of action (nitrofurantoin)

8
Faculty of Pharmacy © Ramaiah University of Applied Sciences
 Urinary excretion studies
Three major parameters examined in urinary excretion data obtained with single dose
study are:
1. Xu Cumulative amount of drug excreted in urine. Directly related to the amount of
drug absorbed; increases as the absorption increases; analogous to AUC
2. (dXu/dt)max Maximum urinary excretion rate. It is analogous to Cmax derived from
plasma level time studies since rate of appearance of drug in urine is proportional to
its concentration in systemic circulation; increases as rate and extent of absorption
increases
3. (tu)max Time for maximum excretion rate. It is analogous to tmax of plasma level
time data. (tu)max decreases as absorption rate increases.

9
Faculty of Pharmacy © Ramaiah University of Applied Sciences
 • Extent of bioavailabilityis calculated from
equation below:-
(Xu∞)oral Div
F= (X u∞)iv Doral
(X u∞)test Dstd
Fr = (X u∞)std Dtest

F= ( Xu, ss )test Dstd τtest

Thus, the bioavailability can be calculated in
(xu,ss )std Dtest τ std the steady state
10
Faculty of Pharmacy © Ramaiah University of Applied Sciences
 Advantages of Urinary Excretion Method

Useful when there is lack of sufficiently sensitive analytical

techniques to measure concentration of drug in plasma

Noninvasive method therefore better subject compliance

Convenience of collecting urine samples in comparison to

drawing of blood periodically

If any case the urine drug concentration is low, assaying of

larger sample volume is relatively more

Direct measurement of bioavailability, both absolute &

relative is possible without the necessity of fitting the data
to the mathematical model

11
Faculty of Pharmacy © Ramaiah University of Applied Sciences
 Acute Pharmacologic Response
• An acute pharmacological effect can be used as an index of drug BA
• Used when bioavailability measurements by pharmacokinetic method is difficult,
inaccurate or non reproducible
• Pharmacodynamic effect is measured over a period of time after drug administration;
Requires demonstration of a dose-response curve
• Effects such as change in ECG, EEG readings or pupil diameter are reflected to the time
course of a given drug.
• BA can determined by construction of pharmacological effect time curve as well as a
dose response curve

12
Faculty of Pharmacy © Ramaiah University of Applied Sciences
 Acute Pharmacologic Response

Pharmacologic response tend to be more variable and accurate

correlation between measured response & drug available from
formulation is difficult

Disadvantages
The observed response may be due to an active metabolite whose
concentration is not proportional

13
Faculty of Pharmacy © Ramaiah University of Applied Sciences
 Therapeutic Response
• Most definite method of all
• Based on observing clinical response to a drug formulation given to patients suffering
from disease for which it is intended to be used.

Drawbacks:
• Quantization of observed response is too improper to allow for reasonable assessment
of relative bioavailability between two dosage forms of same drug
• 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
• 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.
• Many patients receive more than one drug and the results obtained from a BA study
could be compromised because of a drug-drug interaction.

14
Faculty of Pharmacy © Ramaiah University of Applied Sciences
 Clinical observation
• Least accurate, least sensitive and least reproducible

• Well controlled clinical trials in humans establish the safety and effectiveness of drug
products and may be used to assess BA

• Clinical studies have been used to establish bioequivalence for topical antifungals (e.g.
Ketoconazole) and topical acne preparations

• Used only when analytical method and pharmacodynamic methods are not available

15
Faculty of Pharmacy © Ramaiah University of Applied Sciences
 In vitro drug dissolution studies
• Monitoring batch-to-batch consistency through use of in vivo tests is costly, tedious and
time consuming

• In vitro methods are an inexpensive substitute for in vivo methods

• In vitro dissolution tests – a tool to at least quantitatively assure about biological

availability of a drug from its formulation

• In vitro dissolution models aim at mimicking the environment offered by biological

system

16
Faculty of Pharmacy © Ramaiah University of Applied Sciences
 Factors to be considered in design of a dissolution test
• Factors related to apparatus – design, size of container (mL to L), shape (round bottom
or flat), nature of agitation (stirring, rotation or oscillation), speed of rotation,
performance precision of the apparatus etc.

• Factors related to dissolution fluid – composition (water, 0.1N HCl, PB, SGF, SIF),
viscosity, volume (larger than needed to completely dissolve drug), temp. (37 °C),
maintenance of sink (drug conc. is maintained at low level), non-sink conditions (gradual
increase in drug conc)

• Process parameters – introduction of dosage form, sampling techniques, changing the

dissolution fluid etc.
17
Faculty of Pharmacy © Ramaiah University of Applied Sciences
 In vitro dissolution models
Three general categories of dissolution apparatus :

1. Closed-compartment apparatus

2. Open-compartment (continuous flow-through) apparatus

3. Dialysis systems

18
Faculty of Pharmacy © Ramaiah University of Applied Sciences
 In vitro dissolution models

19
Faculty of Pharmacy © Ramaiah University of Applied Sciences