Biological assay of insulin 1

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

Insulin

e standard

Diluting with 0.9%

nacl

30 and 60mili

unitz/ml

REFERNCE SOLUTION:-

Insuline standard

Diluting

with

0.9%

nacl

30 and 60mili

unitz/ml

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 Specific

quantity of insulin USP

Rs

Make a solution

40units/ml(conc.soln)

Make 2 dilutions

SAMPLE SOLUTION Same dilutions are pre prepared

Animal:-24 rabbitRabbit weight 1.8kg-3.0kgMake 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