A new and versatile method for determination of thiolamines of biological importance
A new and versatile method for determination of thiolamines of biological importance
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DOI：
10.1016/s0378-4347(01)00298-5
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发表时间：
2001-09-15
影响因子：
3
通讯作者：
Roberts, JC
中科院分区：
文献类型：
--
作者：
Dominick, PK;Cassidy, PB;Roberts, JC
本文介绍了一种分离和定量测定几种重要生物硫醇胺的方法。该方法采用C-18反相HPLC系统分离还原型和氧化型谷胱甘肽和半胱氨酸的二硝基苯基衍生物，并依赖于内标物N-α-甲基赖氨酸，以尽量减少实验误差。使用几个标准在三种基质（水、HepG 2细胞裂解物和小鼠肝匀浆）中对该方法进行了验证。在10至50 nmol/ml的浓度范围内，谷胱甘肽、谷胱甘肽二硫化物、半胱氨酸和胱氨酸的二硝基苯基衍生物的检测器响应呈线性。测定了日间和日内变异、生物基质中的回收率百分比以及检测限和定量限。为了获得最准确的测定结果，必须每天制作标准曲线，并使用与分析相同的基质。(C)2001 Elsevier Science BM版权所有。
A method for the separation and quantitation of several important biological thiolamines is described. The procedure employs a C-18 reversed-phase HPLC system to separate the dinitrophenyl derivatives of reduced and oxidized glutathione and cysteine and relies on an internal standard, N-epsilon-methyllysine, to minimize experimental error. The method was validated in three matrices (water, HepG2 cell lysates, and mouse liver homogenates) using several criteria. The detector response was linear for the dinitrophenyl derivatives of glutathione, glutathione disulfide, cysteine, and cystine in the concentrations ranging from 10 to 50 nmol/ml. Inter- and intra-day variation, percent recovery in the biological matrices, and limits of detection and quantitation were determined. For the most accurate determination, it is essential that standard curves be produced daily and in the same matrix as that being analyzed. (C) 2001 Elsevier Science BM All rights reserved.