CHAPTER -3
BIOAVAILABILITY STUDIES
BIOAVAILABILITY
• The differences in the bioavailability was observed with different
digoxin formulations in the year 1971.
• Prior to 1971, digoxin was assumed to have consistent effects
across different formulations. However, clinical observations and
pharmacokinetic studies revealed:
• Tablet forms had lower and more variable bioavailability (often
60–80%).
• Elixirs and soft gelatin capsules provided higher and more
predictable absorption (around 90–100%).
Key Impacts of the 1971 Bioavailability Findings:
Therapeutic Inconsistency: Patients experienced unexpected toxicity or lack of
efficacy when switched between formulations.
FDA Involvement: The U.S. FDA began to require strictly followed the
bioequivalence standards for digoxin products, pushing for more uniformity in
performance.
Clinical Practice Shift: Physicians began to choose formulations more carefully
and monitor serum digoxin levels more routinely.
Development of Therapeutic Drug Monitoring (TDM): Digoxin was among the
first drugs for which routine blood level monitoring became standard to
optimize dosing and prevent toxicity.
BIOAVAILABILITY STUDIES
• In approving a drug product for marketing, the FDA ensures
that the drug product is safe and effective for its labeled
indications for use. Moreover, the drug product must meet all
applicable standards of identity, strength, quality, and purity.
To ensure that these standards are met, the FDA requires
bioavailability /pharmacokinetic studies and, where
necessary, bioequivalence studies for all drug products (FDA
Guidance for Industry).
A primary concern in
biopharmaceutics is the
bioavailability of drugs.
Bioavailability
Refers to the measurement of the rate
and extent of active drug that reaches the
systemic circulation.
Means access to the bloodstream
OBJECTIVES
1. Determine the rate and extent of drug absorption.
2. Assess the influence of formulation, route, or other factors (e.g., food).
3. Establish pharmacokinetic parameters like:
o Cmax (maximum plasma concentration)
o Tmax (time to reach Cmax)
o AUC (area under the concentration–time curve)
o t½ (elimination half-life)
Bioavailability
Bioavailability:
It is a measurement of the rate and extent of a therapeutically active drug
that reaches the systemic circulation and is available at the site of action.
Absolute bioavailability:
Absolute bioavailability compares the bioavailability (estimated as area
under the curve, or AUC) of the active drug in systemic circulation following
non-intravenous administration (i.e., after oral, rectal, transdermal,
subcutaneous administration), with the bioavailability of the same drug
following intravenous administration.
ABSOLUTE BIOAVAILABILITY
• The systemic availability of a drug administered orally is determined in
comparison to its iv administration.
• Characterization of a drug's absorption properties from the e.v. site.
F = AUCev / dose
AUCiv / dose
Absolute availability using urinary drug excretion data can be determined
by the following:
Where [D u] ∞ is the total amount of drug excreted in the urine.
Cont…
It is the fraction of the drug absorbed through non-intravenous administration
compared with the corresponding intravenous administration of the same
drug.
RELATIVE BIOAVAILABILITY
• The availability of a drug product as compared to another dosage form or
product of the same drug given in the same dose.
• Characterization of absorption of a drug from its formulation.
• In case of urinary drug excretion data can be determined by the following:
Relative Bioavailability:
This measures the bioavailability (estimated as area under the curve, or
AUC) of a certain drug when compared with another formulation of the
same drug, usually an established standard, or through administration via
a different route.
Guidelines for Bioavailability (BA) Studies
Bioavailability studies are governed by detailed regulatory guidelines to ensure the
quality, safety, and efficacy of pharmaceutical products. These are primarily set by
agencies such as:
1. FDA (U.S. Food and Drug Administration)
2. EMA (European Medicines Agency)
3. WHO (World Health Organization)
4. ICH (International Council for Harmonization)
DESIGN AND CONDUCT OF STUDIES
Reference: FDA Guidance for Industry – Bioavailability and Bioequivalence
Studies Submitted in NDAs or INDs – General Considerations
▪ A bioequivalence study is basically a comparative bioavailability study designed to establish equivalence
between test and reference products.
1. Design:
∙ Typically a single-dose, crossover study in healthy volunteers.
∙ Randomized, two-period, two-treatment, two-sequence.
∙ Fasting and fed conditions may both be required.
2. Subjects:
• Usually healthy adults (18–55 years).
• Number of subjects should be statistically powered (often ≥ 12).
A. Selection of subjects:
Aim to minimize variability and permit detection of differences between pharmaceutical
products.
The studies should normally be performed with healthy volunteers.
They should be screened for suitability by means of clinical laboratory tests, review of medical
history, and medical examination.
B. Standardization of the study:
The test conditions should be standardized in order to minimize the variability of all factors
involved.
Standardization of the diet, fluid intake and exercise is recommended.
The subjects should not take other medicines during a suitable period before and during the
study.
C. Inclusion of patients:
If the investigated active substance is known to have adverse effects and the pharmacological
effects or risks are considered unacceptable for healthy volunteers it may be necessary to use
patients instead, under suitable precautions and supervision.
3. Characteristics to be investigated:
In most cases evaluation of bioavailability and bioequivalence will be based upon the measured
concentrations of the parent compound.
In some situations, measurements of an active or inactive metabolite is carried out.
The plasma concentration versus time curves are mostly used to assess extent and rate of
absorption.
The use of urine excretion data may be advantageous in determining the extent of drug input in
case of products predominately excreted renally.
Specificity, accuracy and reproducibility of the methods should be sufficient.
[Link] analysis:
It is conducted according to the applicable principles of Good Laboratory Practice (GLP).
Determination of the active moiety and/or its metabolite’s in plasma, urine or any other
suitable matrix must be well characterized, fully validated and documented to yield reliable
results that can be reasonably interpreted.
5 Reference and test product:
The choice of reference product should be justified by the applicant and agreed upon by the
regulatory authority.
The test products used in the bio-study must be prepared in accordance with
GMP-regulations.
[Link] analysis:
To quantify the difference in bioavailability between the reference and test products and to
demonstrate that any clinically important difference.
[Link] analysis:
The statistical analysis (e.g. ANOVA) should take into account sources of variation that can be
reasonably assumed to have an effect on the response variable.
Pharmacokinetic parameters derived from measures of concentration, e.g. AUC, Cmax should
be analyzed using ANOVA.
90% Confidence Interval for AUC and Cmax must fall within: 80–125% (for most drugs)
ICH and EMA Guidelines
∙ Similar principles to FDA, but with regional differences.
∙ EMA requires both bioavailability and bioequivalence data when applicable.
∙ ICH M9 guideline addresses biopharmaceutics classification system (BCS)-based biowaivers,
which may exempt some drugs from in vivo BA studies
Methods to Assess Bioavailability:
I. Dissolution at administration or absorption site:
Method of evaluation: Dissolution rate
Example: In vitro method, (water, buffer, artificial gastric fluid, artificial intestinal
fluid, artificial saliva, artificial rectal fluid).
II. Free drug in systemic circulation:
Method of evaluation: [Link] level time profile
[Link] blood level
[Link] to reach peak
[Link] under blood level time curve
Example: In vivo: whole blood, plasma, serum
• The key parameters for determining bioavailability
1. AUC: The AUC is proportional to the total amount of drug reaching the
systemic circulation, and thus characterizes the extent of absorption.
2. Cmax: Gives indication whether drug is sufficiently absorbed systemically
to provide a therapeutic response.
3. Tmax: The Tmax reflects the rate of drug absorption, and decreases as the
absorption rate increases.
III. Quantification of Pharmacologic effect:
Method of evaluation:
[Link] of effect
[Link] of effect
[Link] of effect
Is based on the assumption that a given intensity of response is associated with a
particular drug concentration at the site of action.
The draw backs are difficult to monitoring of pharmacologic data, precision and
reproducibility are also difficult to establish.
Example: There are only a limited number of pharmacologic effects (e.g. heart rate,
body temperature, blood sugar levels and blood pressure, ) that are applicable to this
method.
IV. Clinical response:
Method of evaluation: [Link] clinical blind or double- blind study
[Link] clinical success or failure
Example: In vivo, evaluation of clinical responses
V. Urinary Excretion Data:
Method of evaluation: 1. Cumulative amount of drug excreted in the urine (D u)
2. Rate of drug excretion in the urine (dD u/dt)
3. Time for maximum urinary excretion (t) Peak time of excretion
• This method is base on the evidence that urinary excretion of the
unchanged drug is directly proportional to the plasma concentration of
total drug.
• This technique of studying bioavailability is most useful for those drugs
that are not extensively metabolized prior to urinary elimination.
• The three major parameters examined in urinary excretion data are as
follow:
1.(dXu/dt)max : It gives the rate of appearance of drug in the urine is
proportional to its concentration in systemic circulation. Its value
increases as the rate and extent of absorption increases
2. (tu)max : It is analogous to the of plasma level data, its value decreases
as the absorption rate increases.
3. Xu : It is related to the AUC of plasma level data and increases as the
extent of absorption increases.
Example: In vivo: urine sampling analysis. Used for certain thiazide
diuretics and sulfonamides and for drugs that have urine as the site of
action-for example, urinary antiseptics.
Question #1
• Estimate absolute bioavailability and relative bioavailability
of formulation A & B from both plasma and urine data. What
are the assumptions made in your calculation?
Rout Dose (mg) AUC (mg-hr/L) Amount excreted
(mg)
Iv 500 13.1 332
Oral Drug A 1000 20.9 586
Oral Drug B 1000 19.9 554
Question # 2
• The bioavailability of a new investigational drug was studied in 12
healthy volunteers. Each volunteer received either a single oral tablet
containing 500 mg of the drug (AUC 23.7 mg-hr/L), 5 mL of a pure
aqueous solution containing 500 mg of the drug (AUC 27.8 mg-hr/L)
and a single IV bolus injection containing 200 mg of the drug (AUC 9.7
mg-hr/L). Plasma samples were obtained periodically up to 48 hours
after the dose and assayed for drug concentration. Calculate (a) the
relative bioavailability of the drug from the tablet compared to the
oral solution and (b) the absolute bioavailability of the drug from the
tablet.
