QUANTITATIVE DETERMENATION OF GAMMA-GLUTAMYL TRANSFERASE ( ϒ-GT)

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Presentation transcript:

QUANTITATIVE DETERMENATION OF GAMMA-GLUTAMYL TRANSFERASE ( ϒ-GT) CLS 431

Principle of the method ϒ-Glutamyl-p-nitroanilide + Glycylglycine ϒ-GT ϒ-Glutamyl-glycylglycine + p-nitroaniline Enzyme activity is proportional to the increase in absorbance of nitroaniline at 405 nm.

Clinical significance Found in the kidney, pancreas, liver, brain and prostate. Used in the diagnosis and treatment of hepatobiliary diseases such biliary obstruction, cirrhosis or liver tumours. Acute and chronic pancreatitis with biliary tract involvement. Malignant neoplasm of liver and pancreas alcoholism .

Procedure Wavelength ......................... 405 nm. Cuvette ................................ 1cm. light path constant tempreture .................. 25C Pipette into a cuvette Mix, wait for 30 seconds. Read the absorbance against D.W. and start the stopwatch and read absorbance at 1 minute intervals thereafter for 2 minutes. Calculate the difference between absorbance and the average absorbance differences per minute (Δ A/min) WR(ml) 1.0 Sample (µL) 50

Calculations ΔA/min x 2121 = U/L of ϒ-GT Reference values: Men 6-28 U/L Women 4-18 U/L