Glutathione measurement in human plasma Evaluation of sample collection, storage and derivatization conditions for analysis of dansyl derivatives by HPLC
Glutathione measurement in human plasma Evaluation of sample collection, storage and derivatization conditions for analysis of dansyl derivatives by HPLC
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DOI：
10.1016/s0009-8981(98)00089-8
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发表时间：
1998-07-28
影响因子：
5
通讯作者：
Brown, LAS
中科院分区：
文献类型：
--
作者：
Jones, DP;Carlson, JL;Brown, LAS
尽管使用了明显有效的GSH测量分析方法，但人血浆GSH的文献值差异超过10倍。本研究的目的是开发一种程序，以尽量减少可能导致此类差异的样本采集、处理和储存错误。使用具有丹磺酰基衍生物的荧光检测的HPLC进行定量。结果表明，使用蝶形针和注射器采集血液可减少由于有限溶血而导致的高估，并且使用旨在抑制自氧化和酶降解的保存溶液可定量回收GSH和GSSG。稳定性试验表明，非衍生化的样品在-80度下至少稳定2个月，而丹磺酰衍生物在0-4度下在黑暗中稳定12个月。来自59名健康个体（20-43岁）的结果提供了2.09 +/- 1.14微摩尔的平均（+/-1 SD）GSH值。(C)1998 Elsevier Science B. V.保留所有权利。
Literature values for human plasma GSH vary over 10-fold despite the use of apparently valid analytical procedures for GSH measurement. The purpose of this study was to develop a procedure to minimize error in sample collection, processing and storage that could contribute to such differences. HPLC with fluorescence detection of dansyl derivatives was used for quantification. The results show that collection of blood with a butterfly needle and syringe reduces overestimation due to limited hemolysis and that use of a preservation solution designed to inhibit autooxidation and enzymatic degradation allows quantitative recovery of both GSH and GSSG. Stability tests showed that non-derivatized samples were stable for at least 2 months at - 80 degrees while dansyl derivatives were stable in the dark at 0-4 degrees for 12 months. Results from 59 healthy individuals (20-43 years) provided a mean (+/-1 SD) GSH value of 2.09 +/- 1.14 micromolar. (C) 1998 Elsevier Science B.V. All rights reserved.