Estimation of ALT

ESTIMATION OF ALT(SGPT)

Specimen

  • Serum / plasma

Method of estimation: (IFCC procedure)

Principle

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

Estimation

      • ALT - Alanine Aminotransferase
      • LDH - Lactate dehydogenase

Reagent composition

  • ALT-  (R1)
    • Tris Buffer (pH 7.5) 110mmol/L
    • L-Alanine 600mmmol/L
    • LDH >1500U/L
  • ALT - (R2)
    • a Ketogluterate 16mmol/L
    • NADH 0.24 mmol/L

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 1000ul of working reagent with 100ul 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

  • ALT activity (U/L) = (changes in OD /min) x 1745
Last modified: Wednesday, 30 November 2011, 7:25 AM