Upload
ifsha-akhlaq
View
404
Download
1
Embed Size (px)
Citation preview
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