Principle
Amylase test involves use of a chromogenic substrate Gal G, – a CNP (2-chloro-4-nitrophenyl linked with galactosyl maltoside). The direct action of amylase with this substrate results in the release of more than 90% of 2-chloro-4-nitrophenol, which can be monitored by kinetic assay at 405 nm. The increase in absorbance is directly proportional to the amylase activity in the sample.
Procedure
• Reaction type ………………. Kinetic
• Reaction direction ………………. Increasing
• Wavelength ………………. 405 nm
• Flow Cell temperature ………………. 37° C (+ 0.2°C)
• Zero setting with ………………. Distilled water
• Delay time ………………. 60 seconds
• No. of readings ………………. 4
• Interval ………………. 30 seconds
• Blank absorbance limit ………………. < 0.500 Abs
•Sample volume ………………. 0.02 ml (20)
• Linearity ………………. 2000 IU/
Quality Control
To ensure adequate quality control, it is recommended that each batch should include a normal and 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
Mix thoroughly and transfer the assay mixture immediately to the thermostated cuvette and start the stopwatch simultaneously. Record the first reading at 60 second and subsequently three more readings with 30 seconds interval at 405 nm.
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