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BIOASSAY OF VASOPRESSIN



Basis:


  • Vasopressin anti diuretic hormone of the posterior pituitary gland.

  • It has a potent vasoconstriction + antidiuretic activity.

  • It isused for upper esophageal bleeding (20units/ 15 mints).

  • And also used for diabetes insipidus (5-20units/4hrs)


Desmopressin


  • An analogue of vasopressin

  • Has anti diuretic activity

  • Has no vasoconstrictor activity

  • Used as nasal spray

  • For maintenance therapy in diabetes insipidus


Type of Assay:


2x2 randomized graded response assay.


Reference Standard:


Similar to oxytocin,

Freeze / acetone dried powder in ampoule containing 0.05mg of standard preparation = 1 unit.

1 ampoule contains 8.2 units


Methods of Bioassay:


Official:


  • Rat blood pressure method

  • Antidiuretic activity in ethanol rats


Unofficial:


  • RIA


Method Using Blood Pressure Of Anesthetized Rats:


  • Rats weighing 300gm are anesthetized with 1.75ml/kg of 25% urethane (I.P).

  • It is treated with ganglion blocker like hexamethonium (1mg) to produce a steady base line.

  • External jugular vein cannulated for injection of drug.

  • Internal carotid artery cannulated for recording blood pressure.

  • Doses of references standard + test drug injected in a randomized 2x2 assay.

  • Volume of injection 0.1-0.5ml.

  • Initial dose of reference standard may be tried in the range of 6-10 micro units.

  • Relative potency is calculated.

  • This method can also be used for the assay of Epinephrine & Norepinephrine.


Method Using Anti Diuretic Activity Highly Anesthetized Rats:


  • Rat weighing 250-300 grams are hydrated and anesthetized with oral dose of 5ml/100g of 2.5% ethanol.

  • Urinary bladder cannulated for the collection of urine.

  • Tail vein is cannulated for injection of drugs.

  • Urine output is replaced by oral administration of 0.5% ethanol.

  • Decrease in urine output is taken as a parameter anti diuretic activity.

  • The

  • of standard & test drug are injected for a 2x2 assay & relative potency is calculated.


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