DEFINITION:- Estimation of concentration or

potency of a substance by measurement of

biological response it produces.
i.e. Observation of pharmacological effects on
[1] living tissues, or cells
[2] microorganisms
[3] animals

Also known as BioStandardization.


Chemical assay is not available.
Quantity of sample too small.
Estimate concentration of active principle in
tissue extract e.g..insulin.
Estimate pharmacological activity of unidentified
substance,
Measure drug toxicity,
Dose of drug required to produce therapeutic
effect.

The basic principle of bioassay is to compare
the test substance with the International
Standard preparation of the same.
Find out how much test substance is required
to produce the same biological effect, as
produced by the standard.
The standards are internationally accepted
samples of drugs maintained and
recommended by the Expert Committee of
the Biological Standardization of W.H.O.
Representative of substance serves as basis
for comparative measurement of activity.

In India maintained & distributed by:

Central Drug Research Laboratory , Kolkata.

Central Research Institute , Kasauli.

Central Drug Research Institute Lucknow

[1] Quantal Assays [ Direct end point ]
Elicits an All or None response in different
animals
E.g..
Digitalis induced cardiac arrest in guinea pigs & Cats
hypoglycaemic convulsions in mice.
Calculation of LD50 in mice or rats
[2] Graded Response Assays [mostly on
tissues]
Graded responses to varying doses
Unknown dose response measured on same
tissue



END POINT METHOD
The threshold dose producing a positive effect is
measured (one receiving standard and other the
test)
e.g. bioassay of digitalis in cats.
The cat is anaesthetized,and the drug is slowly
infused into the animal and the moment the heart
stops beating and blood pressure falls to zero,the
volume of fluid infused is noted down.
Conc. Of Unknown= Threshold dose of Std. Conc. of Std.
Threshold dose of Test
Here proportionate increase in responses due to
increase in dose or concentration.

E.g. contraction of smooth muscle for histamine
assay.
I t can be done by following methods..
MATCHING METHOD
BRACKETTING METHOD
GRAPHICAL METHOD
MULTIPLE POINT BIOASSAY
Firstly responses of test is taken & is matched
with response of standard drugs.
Done till a closed matching is observed.
Log-dose-response curve is plotted and the
dose of standard producing the same
response as produced by the test sample is
directly read from the graph.
Three point bioassay:- [2+1 dose assay] two
standard & one test responses are taken down.
Fast & convenient
Procedure [E.g. Ach bioassay]
Log dose response [LDR] curve plotted with varying conc
of std Ach solutions and given test solution
Select two std doses s1& s2 [ in 1:2 dose ratio] from linear
part of LDR [ Let the corresponding response be S1, S2]
Choose a test dose t with a response T between S1 & S2
Record 3 sets data [Latin square: Randomisation reduces
error] as follows
s1 s2 t
t s1 s2
s2 t s1




The mean responses are calculated and plotted
against log-dose and amount of standard
producing the same response as produced by
the test is determined graphically as well as
mathematically:
Conc. Of Unknown=
n1/t antilog { T-S1 log n2/n1} Cs
S2-S1
n1= lower std dose
n2= higher std dose
t = Test dose
S1 & S2 = Response of n1 & n2 respectively.
T = Response of t
Cs = Conc. Of std.
FOUR POINT BIOASSAY
two standards & two test responses are
recorded.
Record 4 sets of data using latin square for s1 s2 t1 &t2
Conc. Of Unknown=



SIX PONT BIOASSAY
Three standards & three test responses are
recorded


ANY QUESTIONS..