使用离心式快速分析仪酶法测定血清或血浆中的乙酸盐。

Enzymatic determination of acetate in serum or plasma using a centrifugal fast analyser.

作者信息

Trivin C, Lenoir F, Bretaudière J P, Sachs C

出版信息

Clin Chim Acta. 1982 May 6;121(1):43-50. doi: 10.1016/0009-8981(82)90209-1.

Abstract

A simple enzymatic spectrophotometric micromethod is described for direct kinetic assay of acetate in serum or plasma using the Eni-Gemsaec centrifugal fast analyser. The method is based on the transformation of acetate and ATP into acetylphosphate and ADP by acetate kinase (EC 2.7.2.1). ADP is further measured by two coupling reactions involving pyruvate kinase (EC 2.7.1.40) and lactate dehydrogenase (EC 1.1.1.27) with measurement of NADH consumption at 340 nm. The method involves a reagent blank for compensation of reagent deterioration, a preincubation of 3 min without acetate kinase to eliminate any interference due to endogenous pyruvate, and a two-point kinetic protocol with measurements of absorbance at 95s and 395 s. The analytical performances of the proposed method were investigated using an evaluation scheme proposed by the French Society of Clinical Biology.

摘要

描述了一种使用Eni-Gemsaec离心快速分析仪直接动力学测定血清或血浆中乙酸盐的简单酶促分光光度微量法。该方法基于乙酸激酶（EC 2.7.2.1）将乙酸盐和ATP转化为乙酰磷酸和ADP。通过涉及丙酮酸激酶（EC 2.7.1.40）和乳酸脱氢酶（EC 1.1.1.27）的两个偶联反应进一步测量ADP，并在340nm处测量NADH的消耗。该方法包括用于补偿试剂变质的试剂空白、在无乙酸激酶的情况下预孵育3分钟以消除内源性丙酮酸的任何干扰，以及在95秒和395秒测量吸光度的两点动力学方案。使用法国临床生物学协会提出的评估方案研究了该方法的分析性能。