ESTIMATION OF SGOT

For the Determination of SGOT (AST) Activity in Serum

1. TOPIC :- ESTIMATION OF SGOT (AST) KIT. Method For the Determination of SGOT (AST) Activity in Serum .

2. Test tube Test tube stand Micropipette Micro Tips Blood sample SGOT kit Syringe Vacutainer tube Tourniquet REQUIRMENTS:-

3. SGOT (AST) catalyzes the transfer of amino group between L- Asparate 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(AST) activity in the sample. PRINCIPLE :

4. L- Asparate + α- ketoglutarate L- Glutamate + oxaloacetate Oxaloacetate + NADH +H+ L- malate + NAD+

5. WORKING REAGENT:- mix 4 parts of R1 + 1 part of R2 Pipette into clean dry test tubes labeled as Standard (S), and Test (T): T Working Reagent:- 1000 μ l (1ml) Specimen :- 100 μ l (0.10m) # Mix well and read the initial absorbance A after 1 min. and repeat 0 the absorbance reading after 1, 2 & 3 minutes at 37oC Procedure:

6. A/min X factor= AST activity( U/L) Factor= 1746 NORMAL VALUES: (Serum) Males : ≤37 U/L Females: ≤ 31 U/L CLINICAL SIGNIFICANCE: INCREASES : Increased levels are found in hepatobiliary disease. Calculation:

7. THANK YOU