基于多酶氧化还原系统抑制作用的硫普罗宁荧光测定法。

Fluorometric assay of tiopronin based on inhibition of multienzyme redox system.

作者信息

Xu Jing, Cai Ruxiu, Wang Jun, Liu Zhihong, Wu Xinguo

机构信息

Department of Chemistry, Wuhan University, China.

出版信息

J Pharm Biomed Anal. 2005 Sep 1;39(1-2):334-8. doi: 10.1016/j.jpba.2005.03.004.

DOI:10.1016/j.jpba.2005.03.004

PMID:16085148

Abstract

In this paper, a simple and sensitive fluorimetric method for the determination of tiopronin (N-(2-mercaptopropionyl)-glycine) is proposed. The method is based on the strong inhibitory effect of tiopronin on the multienzyme redox system of hemoglobin, nicotinamide adenine dinucleotide (NADH) and H(2)O(2), in which the intrinsic fluorescence of NADH was employed as the detection signal. The calibration graph is linear in the range 6.13 x 10(-7) to 6.13 x 10(-6) M with a detection limit of 1.65 x 10(-7) M and the relative standard deviation of 2.02%. Kinetics in the pseudo-first-order conditions was investigated by stopped-flow spectrofluorometry and the inhibition mechanism of tiopronin was verified of the competitive type.

摘要

本文提出了一种简单灵敏的荧光分析法用于测定硫普罗宁（N-(2-巯基丙酰基)-甘氨酸）。该方法基于硫普罗宁对血红蛋白、烟酰胺腺嘌呤二核苷酸（NADH）和H₂O₂的多酶氧化还原系统具有强烈抑制作用，其中利用NADH的固有荧光作为检测信号。校准曲线在6.13×10⁻⁷至6.13×10⁻⁶ M范围内呈线性，检测限为1.65×10⁻⁷ M，相对标准偏差为2.02%。采用停流荧光光谱法研究了伪一级条件下的动力学，并验证了硫普罗宁的抑制机制为竞争型。