Examination OF SGOT

Prepared By: Ayesha Mudassar

Roll No: 20902

Discipline: Medical Laboratory Technology

Serum Glutamic Oxaloacetic Transaminase, an enzyme that is normally present in liver and heart cells.

SGOT is released into blood when the liver or heart is damaged.

The blood SGOT levels are elevated with liver damage viral hepatitis

With heart damage heart attackSome medications can also raise SGOT levels.SGOT is also called Aspartate aminoTransferase

(AST).

Introduction

Principle• SGOT (AST) catalyzes the transfer of amino group

between L-Aspartate and a-Ketoglutarate to form Oxaloacetate and Glutamate. The Oxaloacetate formed reacts with NADH in the presence of Malate Dehydrogenase to form NAD. The rate of oxidation of NADH to NAD is measured as a decrease in absorbance which is proportional to the SGOT (ASAT) activity in the sample.

L-Aspartate a-Ketoglutarate SGOT Oxaloacetate L-Glutamate

Oxaloacetate NADH H+ MDH Malate NAD+

*Procedure

*Wavelength / filter : 340 nm/ filter *Temperature : 37°C / 30°C / 25°C *Light path : 1 cm

Substrate Start Assay Pipette into a clean dry test tube labeled as Test (T):

Mix well and read the initial absorbance A & repeat the absorbance reading after every 1, 2, & 3 minutes.

Calculate the 0 mean absorbance change per minute (D A/min.).

Additional Sequence Temp.250C/300C

Temp.370C

Enzyme Reagent L1 0.8ml 0.8mlSample 0.2ml 0.1mlIncubate at the assay temp. for 1 min. & addStarter Reagent L2 0.2ml 0.2ml

Sample Start Assay Pipette into a clean dry test tube labelled as Test (T):

Mix well and read the initial absorbance A after 1 minute & repeat the absorbance reading after every 1, 2, & 3 minutes.

Calculate the 0 mean absorbance change per minute (D A/ min.).

Additional Sequence Temp.250C/300C

Temp.370C

Working Reagent 1.0ml 1.0ml

Incubate at the assay temp. for 1min. & add

Sample 0.2ml 0.1ml

Calculation & Normal RangeCALCULATIONS:Substrate/ Sample start SGOT (ASAT) Activity in U/L 25°C / 30°C = A min. x 952

37°C = A min. x 1746 NORMAL REFERENCE VALUES:Serum Males: Upto 37 U/L at 37°C Serum Females: Upto 31 U/L at 37°C

Interpretations• Elevated levels of AST I. By liver damage due to conditions like Liver cirrhosis & Hepatitis.II. By decaying tumor or necrosis. • Heart attack• Heart failure • Kidney damage • Lung damage III. Use of medications such as • Antibiotics, Statins, Narcotics, Aspirin, Chemotherapy, and HarbuturesIV. Due to high doses of vitamin AV. Cancer • The AST test is more effective when detecting liver damage caused by

over consumption of alcohol than alanine aminotransferase (ALT) test.

Variations

1. Hemolysis interferes with the test. 2.Triglycerides up to 400 mg/dl. 3.A number of drugs and substances. 4.Bilirubin up to 20 mg/dl, and

hemoglobin up to 400 mg/dl have a negligible effect on this procedure.

5.Patient with severe vitamin B6 deficiency could have a decrease in the recovery of AST presumably due to lack of pyridoxal phosphate.

Reaction: U.V.KineticWavelength: 340nmIncubation Temp. : 370CRead Time: 180 sec.Delay Time: 60 sec.No. of reading: 4Interval: 60Sample Volume: 0.10mlReagent Volume: 1.00mlFactor: 1746Reaction Slope: DecreasingUnits: U/LDirections For Use On

Analyser