Preparation of a New Fluorogenic Substrate of α-Amylases and a Simple α-Amylase Assay by HPLC

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Abstract
A new substrate of α-amylases, O -6-deoxy-6-[(2pyridyl)amino-α-D-glucopyranosyl-(1→4)- O -α-D-glucopyranosl-(1→4)- O -α-D-glucopyranosyl-(1→4)- O -α-D-glucopyranose, was prepared using dextrin as a starting material. Compared with other substrates so far reported, the fluorogenic substrate is unique in that it is resistant to exo-α-glucosidases due to the blocking group introduced into the non-reducing end glucose residue. The product of α-amylase digestion was rapidly separated from the substrate and was detected very sensitively by HPLC and a fluorescence detector. This method for α-amylase assay was also applied for determination of α-amylase in human serum.
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