BIOPHARMACEUTICS

Assessment of Biopharmaceutical properties

(Bioavailability and bioequivalence)
Lecture 7

Dr Omar A Hamid

Department of pharmaceutics

Assessment of biopharmaceutical properties

• Biopharmaceutics is involved with factors that influence the rate and

extent of drug absorption.

• As discussed earlier, these factors may affect :

• the release of a drug from its dosage form,

• its dissolution into physiological fluids,
• its stability within those fluids,
• its permeability across the relevant biological membranes and,
• its presystemic metabolism.
 The key biopharmaceutical properties that can be quantified and
therefore give an insight into the absorption of a drug are its:

• Release from its dosage form into solution at the absorption site;

• Stability in physiological fluids;

• Permeability;

• Susceptibility to presystemic clearance.

• Assessment of bioavailability.

ASSESSMENT OF BIOAVAILABILITY

The measurement of bioavailability gives the net result of the effect of the
release of drug into solution in the physiological fluids, its stability in those
physiological fluids, its permeability and its presystemic metabolism on the
rate and extent of drug absorption by following the concentration time
profile of drug in a suitable physiological fluid.

The most commonly used method of assessing the bioavailability of a drug

involves the construction of a blood plasma concentration-time curve, but
urine drug concentrations can also be used.
Plasm concentration-time curve
 Several parameters based on the plasma concentration-time curve which are
important in bioavailability studies

• Minimum effective (or therapeutic) plasma concentration It is assumed

that some minimum concentration of drug must be reached in the plasma
before the desired therapeutic or pharmacological effect is achieved.

• Maximum safe concentration The concentration of drug in the plasma

above which side-effects or toxic effects occur is known as the maximum
safe concentration.

• Therapeutic range or window A range of plasma drug concentrations is

also assumed to exist, over which the desired response is obtained yet toxic
effects are avoided.

Onset : the time required to achieve the minimum effective plasma concentration
following administration of the dosage form.

Duration: is the period during which the concentration of drug in the plasma
exceeds the minimum effective plasma concentration.

Peak concentration This represents the highest concentration of the drug achieved
in the plasma, which is often referred to as the C Pmax.

Time of peak concentration: This is the period of time required to achieve the
peak plasma concentration of drug after the administration of a single dose. This
parameter is related to the rate of absorption of the drug and can be used to assess
that rate. It is often referred to as the Tmax.
Area under the plasma concentration—time curve This is related to the total
amount of drug absorbed into the systemic circulation following the
administration of a single dose, and is often known as the AUC.

However, changes in the area under the plasma concentration-time curve need
not necessarily reflect changes in the total amount of drug absorbed, but can
reflect modifications in the kinetics of distribution, metabolism and
excretion.

The use of plasma concentration-time curves in bioavailability studies

In order to illustrate the usefulness of plasma concentration-time curves in

bioavailability studies to assess the rate and extent of absorption, the
administration of single equal doses of three different formulations, A, B and
C, of the same drug to the same healthy individual by the same route of
administration on three separate occasions can be considered.

The plasma concentration time profiles for the three formulations are shown
in the following figure:
The differences between the three curves are attributed solely to differences in
the rate and/or extent of absorption of the drug from each formulation. The
plasma profiles show:

1- Each of the three formulations (A, B and C) results in a different peak

plasma concentration. However, the areas under the curves for formulation A
and B are similar, and this indicates that the drug is absorbed to a similar extent
from these two formulations.

2- The times at which the peak plasma concentrations occur for formulations A
and B show that the drug is absorbed faster from A than from B, meaning that
formulation A shows a fast onset of therapeutic action, but as its peak plasma
concentration exceeds the maximum safe concentration it is likely that this
formulation will result in toxic side effects.
3- Formulation B, which gives a slower rate of absorption than A, shows a
slower therapeutic onset than A, but its peak plasma concentration lies within
the therapeutic range. In addition, the duration of action of the therapeutic effect
obtained with formulation B is longer than that obtained with A. Hence
formulation B appears to be superior to formulation A from a clinical viewpoint,
in that its peak plasma concentration lies within the therapeutic range of the
drug and the duration of the therapeutic effect is longer.

4- Formulation C gives a much smaller area under the plasma concentration-

time curve, indicating that a lower proportion of the dose has been absorbed.
Together with the slow rate of drug absorption from formulation C, results in the
peak plasma concentration not reaching the minimum effective concentration,
i.e. formulation C does not produce a therapeutic effect and consequently is
clinically ineffective as a single dose

It is important to realize that the study of bioavailability based on drug

concentration measurements in the plasma (or urine) is complicated by the fact
that such concentration-time curves are affected by factors other than the
biopharmaceutical factors of the drug product itself. Other factors can
complicate the interpretation of the bioavailability studies, such as

• body weight,
• sex and age of the test subjects,
• disease states,
• genetic differences in drug metabolism, excretion and distribution, food and
water intake,
• concomitant administration of other drugs,
• stress
• time of administration of the drug.
Absolute and relative bioavailability

Absolute bioavailability
The absolute bioavailability of a given drug from a dosage form is the fraction
(or percentage) of the administered dose which is absorbed intact into the
systemic circulation.

Absolute bioavailability may be calculated by comparing the total amount of

intact drug that reaches the systemic circulation after the administration of a
known dose of the dosage form via a route of administration, with the total
amount that reaches the systemic circulation after the administration of an
equivalent dose of the drug in the form of an intravenous bolus injection.
.

An intravenous bolus injection is used as a reference to compare the systemic

availability of the drug administered via different routes, because it has no
absorption barriers to cross and is therefore considered to be totally
bioavailable.
The absolute bioavailability of a given drug using plasma data may be
calculated by comparing the total areas under the plasma concentration-time
curves obtained following the administration of equivalent doses of the drug
via an absorption site (AUCT)abs and via the intravenous route (AUCT)iv in the
same subject on different occasions.

𝐴𝑈𝐶𝑇 𝑎𝑏𝑠
Absolute bioavailability =
𝐴𝑈𝐶𝑇 𝑖𝑣
If different doses of the drug are administered by both routes, a correction for the
sizes of the doses can be made as follows:
𝐴𝑈𝐶𝑇 𝑎𝑏𝑠/𝐷𝑎𝑏𝑠
Absolute bioavailability =
𝐴𝑈𝐶𝑇 𝑖𝑣 /𝐷𝑖𝑣
where Dabs is the size of the single dose of drug administered via the absorption
site, and Div is the size of the dose of the drug administered as an intravenous
bolus injection.
 Measurements of absolute bioavailability obtained by administering a given drug
in the form of a simple aqueous solution by both the oral and the intravenous
routes provide an insight into the effects that factors associated with the oral
route may have on bioavailability, e.g. presystemic metabolism by the intestine or
liver, the formation of complexes between the drug and endogenous substances (e.g.
mucin) at the site of absorption and drug stability in the gastrointestinal fluids.

Relative bioavailability
In the case of drugs that cannot be administered by intravenous bolus injection,
the relative (or comparative) bioavailability is determined rather than the
absolute bioavailability.

In this case the bioavailability of a given drug from a 'test' dosage form is
compared to that of the same drug administered in a 'standard' dosage form,
which is either an orally administered solution (from which the drug is
known to be well absorbed) or an established commercial preparation of
proven clinical effectiveness.

Hence relative bioavailability is a measure of the fraction (or percentage) of a

given drug that is absorbed intact into the systemic circulation from a dosage
form relative to a recognized (i.e. clinically proven) standard dosage form of
that drug.
The relative bioavailability of a given drug administered as equal doses of a test
dosage form and a recognized standard dosage form, respectively, by the same
route of administration to the same subject on different occasions, may be
calculated from the corresponding plasma concentration-time curves as follows:

𝐴𝑈𝐶𝑇 𝑡𝑒𝑠𝑡
Relative bioavailability =
𝐴𝑈𝐶𝑇 𝑠𝑡𝑎𝑛𝑑𝑎𝑟𝑑

When different doses of the test and standard dosage forms are administered, a
correction for the size of dose is made as follows:

𝐴𝑈𝐶𝑇 𝑡𝑒𝑠𝑡/𝐷𝑡𝑒𝑠𝑡
Relative bioavailability =
𝐴𝑈𝐶𝑇 𝑠𝑡𝑎𝑛𝑑𝑎𝑟𝑑 /𝐷𝑠𝑡𝑎𝑛𝑑𝑎𝑟𝑑

Relative bioavailability measurements are often used to determine the effects

of dosage form differences on the systemic bioavailability of a given drug.

Numerous dosage form factors can influence the bioavailability of a drug.

These include the type of dosage form, differences in the formulation of a
particular type of dosage form, and manufacturing variables employed in
the production of a particular type of dosage form.
 Bioequivalence

An extension of the concept of relative bioavailability, which essentially

involves comparing the total amounts of a particular drug that are absorbed
intact into the systemic circulation from a test and a recognized standard
dosage form,

Bioequivalence is the determining whether test and standard dosage

forms containing equal doses of the same drug are equivalent or not in
terms of their rates and extents of absorption (i.e. systemic
availabilities). This is called bioequivalence.

Two or more chemically equivalent products (i.e. products containing equal

doses of the same therapeutically active ingredient (s) in identical types of
dosage form are said to be bioequivalent if they do not differ significantly in
their bioavailability characteristics when administered in the same dose
under similar experimental conditions.

Two or more chemically equivalent drug products may be considered

bioequivalent if there is no significant difference between any of the following
parameters:

• maximum plasma concentrations (Cmax),

• time to peak height concentration (Tmax) and
• areas under the plasma concentration-time curves (AUC).
• Bioequivalence studies are therefore important in determining whether
chemically equivalent drug products manufactured by different companies
are therapeutically equivalent, i.e. produce identical therapeutic responses in
patients.

The problem is how much of a difference can be allowed between two

chemically equivalent drug products and still permit them to be considered
bioequivalent. Should this be 10%, 20%, 30% or more? The magnitude of the
difference that could be permitted will depend on the significance of such a
difference on the safety and therapeutic efficacy of the particular drug. This
will depend on such factors as the toxicity, the therapeutic range and the
therapeutic use of the drug.

In the case of a drug with a wide therapeutic range, the toxic effects of which
occur only at relatively high plasma concentrations, chemically equivalent
products giving quite different plasma concentration-time curves (the following
figure) may still be considered satisfactory from a therapeutic point of view,
although they are not strictly bioequivalent.