I prepare these notes for the help of students in study.

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 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.

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 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

7.  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

8. 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.

9.  There are 3 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 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

12.  Fatty change in the liver
 Alcohol abuse
 Cirrhosis
 Mononucleosis
 Drugs (i.e. statins, aspirin, barbiturates, HIV
medication, herbs).

13.  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!

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