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Clinical Chemistry 41: 519-522, 1995;
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Clinical Chemistry, Vol 41, 519-522, Copyright © 1995 by American Association for Clinical Chemistry

Automated measurement of alpha-amylase isoenzymes with 6(3)- deoxymaltotriose as selective amylase inhibitor

R Uchida, S Tokutake, Y Motoyama, K Hosoi and N Yamaji
Research and Development Division, Kikkoman Corp., Chiba, Japan.

We developed an automated method for measurement of alpha-amylase isoenzymes in serum by a single kinetic assay (SKA) and a double kinetic assay (DKA) with 2-chloro-4-nitrophenyl-6(5)-azido-6(5)-deoxy- beta-maltopentaoside as a substrate and 6(3)-deoxymaltotriose (DOG3) as a novel selective amylase inhibitor. DOG3 showed a large difference in inhibitory activity between human pancreatic alpha-amylase (HPA; 86.9% inhibition) and salivary alpha-amylase (32.1% inhibition) at 0.33 mmol/L. Constant inhibition was obtained immediately after addition of DOG3. The inhibitory effect did not change with variation in concentrations of amylase up to approximately 3000 U/L. The results obtained by SKA correlated well with those obtained by three methods: monoclonal antibody (r = 0.988), wheat germ inhibitor (r = 0.989), and DKA (r = 0.995). The within-run and between-run CVs for HPA were 0.63- 2.32% on SKA, 0.69-1.81% on DKA. No significant interferences by endogenous serum compounds were observed with the proposed methods.




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