1. BIOLOGICAL ASSAY OF INSULIN
IFSHA AKHLAQ

2. DEFINATION
 Human insulin is a 2 chain pepitide having the
structure of the antidiabetic harmone produced by
human pancreas.

3. UNITS
 0.0347mg of human insulin is equivalent to 1 IU of
insulin.

4. PRODUCTION
 By enzymatic modification
 By suitable purification of insulin

5. 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

6. 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

7. HYPOGLYCEMIC SEIZURES IN MICE
 SAMPLE SOLUTION PREPARATION
Insuline
standard
Diluting
with 0.9%
nacl
30 and 60mili
unitz/ml

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

9. PROCEDURE
96 mice weighing 20 5g
Randomly distributed into 4 groups
Deprived of food 2-20hr
Inject solution subcutaneously 0.5ml/20g

10. 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

11. RABBIT SUGAR METHOD
 Standard stock solution:-
Dissolve
Specific
quantity of
insulin USP
Rs
Make a
solution
40units/ml
(conc.soln)
Make 2
dilutions

12. 1 unit/ml 2unit/ml
Make dilutions from 40units/ml solution
Dilution no 1 Dilution no 2

13. SAMPLE SOLUTION
 Same dilutions are pre prepared
Animal:-
24 rabbit
Rabbit weight 1.8kg-3.0kg
Make 4 groups each group contain 6 rabbit

14.  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

16.  Take sample nd perform quantatively test
 120-150mg/dl

17. 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