Biological assay of insulin 1

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

8. Refernce solution:-

9. Procedure

10. Rabbit Sugar method Standard stock solution:-

11. Sample solution Same dilutions are pre prepared Animal:- 24 rabbit Rabbit weight 1.8kg-3.0kg Make 4 groups each group contain 6 rabbit

12. 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

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

14. Referances Drug Discovery and Evalution pharmacological Assays 3 rd edition edited by Hans Gerhard Vogal page no 1355 British pharmacopoeia 2011 volume I and II monographs medicinal and pharmaceutical substances