Bioassay of Insulin

• Bioassay of Insulin: Ensures potency, efficacy, and standardization of insulin preparations.
• Bioassay of Insulin: Measures hypoglycemic activity using rabbit or mouse blood glucose levels.

Principle of Bioassay of Insulin

• Insulin lowers blood glucose levels by promoting glucose uptake in peripheral tissues and inhibiting hepatic glucose output.
• The bioassay evaluates the hypoglycemic effect of a test preparation vs. a standard insulin preparation.

Methods

1. Rabbit Method (Hypoglycemic Convulsion Method)

• Procedure:
  1. Preparation: Rabbits fasted for 18–24 hours (water allowed).
  2. Dosing: Standard insulin administered intravenously or subcutaneously; test insulin given to a separate group.
  3. Observations: Monitor for hypoglycemic convulsions, coma, or measure blood glucose levels at intervals.
  4. Potency Determination: Compare ED50 values or onset time for hypoglycemic symptoms between test and standard samples using dose-response curves.
• Potency Determination: Compare the time to convulsion/coma or reduction in blood glucose for standard vs. test.

2. Mouse Blood Glucose Method:

• Procedure:
  1. Preparation: Mice fasted for 6–8 hours.
  2. Dosing: Graded doses of standard insulin and test insulin administered.
  3. Observations: Measure
  4. Data Analysis: Compare dose-response curves for test and standard insulin to determine potency.
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• Potency Determination: Plot the dose-response for blood glucose drop against the standard and test preparations.

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