SGPT
INTENDED USE:
This reagent kit is intended for in-vitro quantitative determination of SGPT (ALT) activity in serum.
PRINCIPLE:
SGPT (ALT) catalyzes the transfer of amino group between L-alanine and α-ketoglutarate to form pyruvate and glutamate. The pyruvate formed reacts with NADH in the presence of Lactate Dehydrogenase (LDH) to form lactate and NAD. The rate of oxidation of NADH to NAD is measured as a decrease in absorbance which is proportional to the SGPT (ALT) activity in the sample.
REACTION:
Alanine aminotransferase
L-Alanine + α-Ketoglutarate → Pyruvate + L-Glutamate
Lactate Dehydrogenase
Pyruvate + NADH + H⁺ → Lactate + NAD⁺
CONTENTS:
Reagent 1: SGPT Enzyme Reagent
Reagent 2: SGPT Substrate Reagent
MATERIALS REQUIRED BUT NOT PROVIDED:
• Clean & Dry Glassware
• Micropipettes & Tips
• Bio-Chemistry Analyzer
SAMPLES:
Serum free of hemolysis. SGPT (ALT) is reported to be stable in serum for 3 days at 2–8°C.
PROCEDURE:
Pipette into clean dry test tubes as (T):
| Addition sequence | Blank | Test |
|---|---|---|
| Working Reagent | 1000 µl | 1000 µl |
| Sample | — | 100 µl |
Mix well and read the initial absorbance after 1 min and repeat the absorbance reading after every 1 min. Calculate the mean absorbance change per minute (ΔA/min).
CALCULATION:
SGPT activity (U/L) = ΔA/min × 1746