kinetic Assay of Chymotrypsin with an Ammonia Gas-Sensing Electode

  • TAGAMI SHOICHIRO
    Faculty of Pharmaceutical Sciences, Toyama Medical and Phrmaceutical University
  • WAKATA TATSURO
    Faculty of Pharmaceutical Sciences, Toyama Medical and Phrmaceutical University

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Description

A simple kinetic method for chymotrypsin assay is described. The based upon the hydrolysis of N-benzoly-L-tyroshinamide as a substrate, resulting in the production of ammonia. The initial rate of ammonia evolution is selectively measured potentiometrically with an ammonia gas-sensing electrode (Horiba ammonia gas-sensing electrode 5002A-06T). Optimal conditions for the assay are specificed. The initial rate of ammonia evolution is proportional to the enzyme activity, and a working range of 0.08 to 0.32 U/ml chymorypsin was obtained. The recovery of chymotrypsin was 99.2-102.5%. In order to study the applicability of the method, recovery experiments were performed by used of an ophthalmic chymotrypsin preparation.

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