Examination OF SGPT

Prepared By:Ayesha Mudassar

Discipline:Medical Laboratory Technology

Introduction

Alanine Aminotransferase (ALT) is an enzyme found primarily in the liver and kidney. It was originally referred to as Serum Glutamic Pyruvic Transaminase (SGPT). 

 Normally, a low level of ALT exists in the serum.

ALT is increased with liver damage and is used to screen or monitor liver disease.

PrincipleSGPT converts L-Alanine and a-Ketoglutarate

to Pyruvate and Glutamate. The Pyruvate formed reacts with 2,4,Dinitrophenyl hydrazine to produce a hydrazone derivative, which in an alkaline medium produces a brown coloured complex whose intensity is measured.

• L-alanine + a-Ketoglutarate SGPT Pyruvate + L-GlutamatepH 7.4

• Pyruvate + 2,4,DNPH Alkaline 2,4,Dinitrophenyl HydrazoneMedium (Brown Coloured Complex)

Procedure One Reagent Procedure:

Mix and after 1min incubation, measure the change of Optical Density per min( OD/min) during 3 minutes.

Working Reagent 1ml

Sample 0.1ml

Two Reagent Procedure:

Mix and after 1min incubation, measure the change of Optical Density per min ( OD/min) during 3 minutes.

Working Reagent 1ml

Sample 0.125ml

Mix, wait 1min and add

Reagent 0.250ml

Calculations & Normal Range

At 340nm with one-reagent procedure and the two-reagent procedure: Activity (U/L) = OD/min 1746.

The reference range for ALT is 20-60 IU/L.

INTERPRETATION There are 3 basic interpreting

conditions: High Levels of ALT. Moderately high levels of ALT. Slightly high levels of ALT.

High levels Of ALT1. Liver damage (acute viral hepatitis,

toxins/drugs including acetaminophen overdose, acute fulminant hepatitis).

2. Peak levels do not necessarily correlate with prognosis.

3. Tumor necrosis.

Moderately High Levels Of ALT Chronic liver disease Alcohol abuse Cholestasis Heart damage (heart attack, heart failure) Kidney damage Muscle injury Hemolysis Heatstroke (level dependent on extent of

tissue damage) High consumption of vitamin A

Slightly High Levels Of ALT Fatty change in the liver Alcohol abuse Cirrhosis Mononucleosis Drugs (i.e. statins, aspirin, barbiturates,

HIV medication, herbs).

Serum, Heparinized or EDTA plasma free from hemolysis.

Hemolysis should be avoided, since ALAT activity in erythrocytes is 3 to 5 times higher than in normal serum.

ALAT activity remains stable in specimen up to 3 days at 15-25ºC or up to 7 days at 2-8ºC.

Although samples frozen at -20ºC can be stored longer than 7 days, it is recommended to perform the assay with freshly collected samples!

Variations

Wave Length : 340nm Incubation Temp : 370 C Incubation Time : 1 min. Read Time : 3 min No. of Readings : 4 Interval Time : 1 min. Sample Volume : 0.1 ml Reagent Volume : 1 ml Unit : U/L Factor : 1746

Directions For Use On Analysers