BIOLOGICAL ASSAY OF INSULIN

IFSHA AKHLAQ
DEFINATION
 Human insulin is a 2 chain pepitide having the
structure of the antidiabetic harmone produced by
human pancreas.
UNITS
 0.0347mg of human insulin is equivalent to 1 IU of
insulin.
PRODUCTION
 By enzymatic modification
 By suitable purification of insulin
CHARACTERS
 Appearance
white powder
 Solubility

insoluble in ethanol, methanol and aqueous

solution at pH 5.1
 Insulin contain

51 amino acid
α chain 21
β chain 30
IDENTIFICATION TEST
 CHEMICAL ANALYSIS
To measure insulin potency quantitatively
liquid chromatography performed
Results
principal peak in the chromatogram
by comparing test solution and referanc
solution
HYPOGLYCEMIC SEIZURES IN MICE

 SAMPLE SOLUTION PREPARATION

Diluting
Insuline with 0.9%
30 and 60mili
standard unitz/ml
nacl
REFERNCE SOLUTION:-

Diluting
Insuline with 0.9% 30 and 60mili
standard unitz/ml
nacl
PROCEDURE

96 mice weighing 20 5g

Randomly distributed into 4 groups

Deprived of food 2-20hr

Inject solution subcutaneously 0.5ml/20g

 Kept at uniform temperature between 29-35°

Take mice in transparent cntainer within an air incubator

Observed for 1.5 hr and no of mice that are dead

RABBIT SUGAR METHOD
 Standard stock solution:-

Dissolve Make a
Specific solution Make 2
quantity of 40units/ml dilutions
insulin USP
Rs (conc.soln)
Make dilutions from 40units/ml solution

Dilution no 1 Dilution no 2

1 unit/ml 2unit/ml
SAMPLE SOLUTION
 Same dilutions are pre prepared
Animal:-
24 rabbit
Rabbit weight 1.8kg-3.0kg
Make 4 groups each group contain 6 rabbit
 Rabbit in fasting condition (18 hr)
 Perform pretest on rabbit

 1unit of insulin(S/C)

 Wait for 5min

 If any convulsion occur then discard the rabbit

 Take sample nd perform quantatively test
 120-150mg/dl
REFERANCES
 Drug Discovery and Evalution pharmacological
Assays 3rd edition edited by Hans Gerhard Vogal
page no 1355
 British pharmacopoeia 2011 volume I and II

monographs medicinal and pharmaceutical

substances