Examination of sgpt

Examination of sgpt

• 2.
   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.
• 4.
  Principle  SGPT convertsL-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.
• 5.
  • L-alanine +a-Ketoglutarate SGPT Pyruvate + L-Glutamate pH 7.4 • Pyruvate + 2,4,DNPH Alkaline 2,4,Dinitrophenyl Hydrazone Medium (Brown Coloured Complex)
• 6.
  Procedure  One ReagentProcedure:  Mix and after 1min incubation, measure the change of Optical Density per min( OD/min) during 3 minutes. Working Reagent 1ml Sample 0.1ml
• 7.
   Two ReagentProcedure:  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
• 8.
  Calculations & NormalRange  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.
• 9.
   There are3 basic interpreting conditions:  High Levels of ALT.  Moderately high levels of ALT.  Slightly high levels of ALT.
• 10.
  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.
• 11.
   Chronic liverdisease  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
• 12.
   Fatty changein the liver  Alcohol abuse  Cirrhosis  Mononucleosis  Drugs (i.e. statins, aspirin, barbiturates, HIV medication, herbs).
• 13.
   Serum, Heparinizedor 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!
• 14.
   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