Veterinary Clinical Biochemistry and Laboratory Diagnosis

Estimation of Aspartate amino transferase

ESTIMATION OF AST (SGOT)

Principle

• Kinetic determination of AST activity based on the following reaction.

• AST --- Aspatate Aminotransferase
• MDH --- Malate dehydogenase

Reagent composition

• AST - (R1)
  • Tris Buffer (pH 7.8) - 88mmol/L
  • L-Aspartate - 260mmmol/L
  • LDH > 1500U/L
  • MDH > 900 U/L
• AST - (R2)
  • a Ketogluterate - 12mmol/L
  • NADH - 0.24 mmol/L

Method : (IFCC Procedure)

1. Mix four volumes of Reagent (R1) with one volume of Reagent (R2). The working reagent is stable for 30 days at 2-8º c
2. Mix 1000 µl of working reagent with 100 µl of serum and incubate for one minute at 37 ° C.
3. Read the changes in absorbance per 20 sec, during three minute using absorbance spectrophotometer

Calculation

• AST activity (U/L) = ------- (changes in OD /min) x 1745