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Examination of sgpt

Alanine aminotransferase (ALT) is an enzyme found primarily in the liver and kidney that was originally referred to as serum glutamic pyruvic transaminase (SGPT). ALT levels are normally low in the serum but increase with liver damage and are used to screen for and monitor liver disease. The ALT test works by measuring the conversion of L-alanine and α-ketoglutarate to pyruvate and glutamate by SGPT, and then measuring the intensity of the brown colored complex formed between pyruvate and 2,4-dinitrophenyl hydrazine. Elevated ALT can indicate liver damage from viral hepatitis, toxins, drugs, tumors, or chronic liver

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 AlanineAminotransferase (ALT) is an enzyme found primarily in the liver and kidney. It was originally referred to as Serum Glutamic PyruvicTransaminase (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.
Principle  SGPT 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-Glutamate pH 7.4 • Pyruvate + 2,4,DNPH Alkaline 2,4,Dinitrophenyl Hydrazone Medium (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.
 There are 3 basic interpreting conditions:  High Levels of ALT.  Moderately high levels of ALT.  Slightly high levels of ALT.
1. 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.
 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
 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!
 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