Comparison of Recent Methods for Amylase Activity Measurement Using Synthetic Substrates

Recently, several methods for measuring the amylase activity have been developed using new synthetic substrates. These methods can detect released chromophores such as 4-nitrophenol (PNP), 2-chloro-4-nitrophenol (CNP), and NADH. We compared ten such methods with the blue dye-linked starch polymer (Blue Starch) method. The intra-run precision (CV) of each method, 0.4-3.3%, was acceptable. The influence from coexisting materials was negligible in each method, and the correlation among the methods was good. There was no problem with the stability of any reagents after preparation. Comparing the reactivities of pancreatic and salivary amylases as a ratio (P/S), the differences among methods became obvious.