VLD 421: Veterinary Clinical Biochemistry and Laboratory Diagnosis-II (0+1)

Estimation of Gamma Glutamyl Transferase

METHOD OF ESTIMATION OF GAMMA GLUTAMYL TRANSFERASE

Sample: Serum ( free of haemolysis)

Method of Estimation : Szasz Method (Agappe Diagnostics )

Principle

• Kinetic determination of Gamma GT according to the following reaction.

GLUPA-C +Glycyglycine      ^{Gamma GT}    L- Gamma- Glutamyl Glycyglycine       + 5-Amino -2 -nitrobenzoic acid. 

GLUPA-C : L- Gamma-Glutamyl - 3 - Carboxy-p - nitroanilide

Reagent composition

• Gamma GT- R1
  • Tris Buffer, (pH 8.25)   -133 mmol/L
  • Glycyglycine                  -   138 mmol/L
• GammaGT -  R2
  • Glupa -C                         - 23 mmol/L10000 U/L

Laboratory procedure

Working reagent : 1000μL

Sample : 100μL

Mix and incubate for 1 min. at 37 ° C. Read the change in absorbance per minute ( OD/min) during 3 minutes..

Calculation

       Gamma GT activity (IU/L) = (Change in OD / min) x 1158