PRINCIPLE
Amylase test involves use of a chromogenic substrate Gal G2 - CNP (2-chloro-4-nitrophenyl linked with galactosylmaltoside). The direct action of amylase with this susbtrate 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 sample.
PREPARATION OF WORKING SOLUTION
DO NOT FREEZE THE REAGENT. Contamination of the reagent should be strictly avoided.
COMPONENTS & CONCENTRATION OF WORKING SOLUTION
Component Concentration
Component | Concentration |
Buffer, pH 6.0 | 50 mmol/| |
Gal G, -CNP | 2.60 mmol/l |
KSCN | 140 mmol/| |
Activators & Stabilizers |
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SPECIMEN COLLECTION & PRESERVATION
Blood should be collected in a clean dry container. Avoid the use of plastic or siliconized container as it may prolong clotting time. Although serum is preferred, plasma with heparin can be used. EDTA, Oxalate or Citrate inhibit the amylase activity and hence cannot be used. Amylase activity is stable in serum for 20 days at 2-8 Degree C.
PROCEDURE
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