Principle
α-ketoglutarate reacts with L-alanine in presence of GPT (ALT) to form pyruvate and Glutamate. The increase in pyruvate is determined in an indicator reaction catalyzed by lactate dehydrogenase. The conversion of NADH to NAD at 340 nm. is proportional to the activity of GPT (ALT) in Serum Plasma and is determined kinetically as rate of decrease in absorbance.
Procedure
Reaction type …………..UV – Kinetic
Reaction direction ………. Decreasing
Wavelength ……………… 340 nm.
Flow Cell temperature….. 37°C
Zero setting with …….. Distilled water
Delay time ……………….. 60 seconds
No. of readings …..4
Interval…… 30 seconds
Blank absorbance limit ……………. 20.900 Abs.
Sample volume ……… 0.05 ml (50 ml)
Working solution volume (4 R, : 1 R.) ……….. 1.0 ml (1000 ul)
Factor ……… 3339
Linearity …………….. 600 IU/I
Quality Control
To ensure adequate quality control, it is recommended that each batch should include normal and an abnormal commercial reference control serum. It should be realized that the use of quality control material checks both instrument and reagent functions together. Factors which might affect the performance of this test include proper instrument function, temperature control, cleanliness of glassware and accuracy of pipetting.
Other Details
The reagent kit should be stored at 2 – 8°C and is stable till the expiry date indicated on the label. R, and P, reagents are stable till expiry at 2- 8°C. The working solution (4 R, +1 R,) is stable for 30 days at 2- 8°C.
Modini Sharma –
We are using accurex products last 2 year ,
Good service and good reagents