Biological assay of insulin 1

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Biological assay of insulin 1

  1. 1. BIOLOGICAL ASSAY OF INSULIN IFSHA AKHLAQ
  2. 2. DEFINATION  Human insulin is a 2 chain pepitide having the structure of the antidiabetic harmone produced by human pancreas.
  3. 3. UNITS  0.0347mg of human insulin is equivalent to 1 IU of insulin.
  4. 4. PRODUCTION  By enzymatic modification  By suitable purification of insulin
  5. 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. 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. 7. HYPOGLYCEMIC SEIZURES IN MICE  SAMPLE SOLUTION PREPARATION Insuline standard Diluting with 0.9% nacl 30 and 60mili unitz/ml
  8. 8. REFERNCE SOLUTION:- Insuline standard Diluting with 0.9% nacl 30 and 60mili unitz/ml
  9. 9. PROCEDURE 96 mice weighing 20 5g Randomly distributed into 4 groups Deprived of food 2-20hr Inject solution subcutaneously 0.5ml/20g
  10. 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. 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. 12. 1 unit/ml 2unit/ml Make dilutions from 40units/ml solution Dilution no 1 Dilution no 2
  13. 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. 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. 15.  Take sample nd perform quantatively test  120-150mg/dl
  16. 16. 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

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