摘要 |
A method and unitized test composition is described for the estimation of an anilide in which the enzymatic hydrolysis of the anilide and colorimetric quantitation of aniline or aniline derivative can be done simultaneously. The hydrolysis of the anilide is catalyzed by a known enzyme, arylacylamidase, E. C. 3.5.1.13. Stabilization of the enzyme is provided by the addition of controlled amounts of a compound containing alcoholic and/or aromatic groups such as ortho-cresol, isopropanol or benzoate. Basically, the unitized test composition comprises (i) arylacylamidase, (ii) a controlled amount of an organic compound containing alcoholic and/or aromatic groups which acts as both a stabilizer for the arylacylamidase and forms a colored product with aniline, and (iii) a novel oxidant/catalytic agent for accelerating color development. The benefit of such methodology results in a one step addition of the complete reagent to the sample, serum or matrix containing the anilide as opposed to a several step addition. The consequence of this is that only one reagent channel is required to perform the test in an automated analyzer as opposed to the usual requirement that several channels be utilized. This method also makes possible the development of a solid-phase test device such as a dip and read test strip containing all the reagents necessary for the test of an anilide. In addition, a method for the stabilization of arylacylamidase enzyme is described.
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