Substrate soln. for carboxylic ester hydrolase determn. - comprising vinyl ester, buffers, salts and nonionic detergent

摘要

A reagent for analysis of carboxylic acid ester hydrolases comprises a water insoluble vinyl ester of a carboxylic acid, a non-ionic detergent of HLB 13-18, a buffer, and neutral salts. The HLB of the detergent and the ionic strength of the soln. are chosen so as to optimise the specific activity of the enzyme; these quantities, and the amts. of the components, are also chosen so that, the mixt. of substrate with enzyme is clear and photometrically stable. The compsns. can be made specific to a particular enzyme in the presence of others, which is useful in analysis e.g. of blood serum, which may contain lipases from several sources. The compsns. are shelf stable and sensitive to low enzyme concns. in short times, and give linear correlation of activity with enzyme concentration. A typical compsn. comprises 5-15 mM vinyl omega-phenyl-octanoate, 5-150 mM polyoxyethylated tert. octyl phenol, 0.05-0.5 pyrophosphate buffer, and 0-25 mM deoxycholate and 0-15 mMcetyldimethylbenzyl ammonium chloride as enzyme activators.