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
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
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
15.
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