Pitfalls in the analysis of the physiological antioxidant glutathione (GSH) and its disulfide (GSSG) in biological samples: An elephant in the room.
Pitfalls in the analysis of the physiological antioxidant glutathione (GSH) and its disulfide (GSSG) in biological samples: An elephant in the room.
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DOI：
10.1016/j.jchromb.2016.02.015
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发表时间：
2016-04-15
期刊：
影响因子：
--
通讯作者：
Rossi R
中科院分区：
文献类型：
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作者：
Giustarini D;Tsikas D;Colombo G;Milzani A;Dalle-Donne I;Fanti P;Rossi R
谷胱甘肽（GSH）是细胞内最丰富的低分子量硫醇，也是体液中主要的抗氧化化合物之一。在促氧化条件下，两个 GSH 分子各提供一个电子并转化为二硫化谷胱甘肽 (GSSG)。 GSH/GSSG 摩尔比被认为是氧化应激和疾病风险的有力指标。尽管人们对 GSH/GSSG 滴定作为硫醇氧化还原平衡的测量方法很感兴趣，但对于生物样品中这些分子的定量的最佳预分析和分析方法尚未达成广泛一致。因此，不同实验室测得的 GSH 和 GSSG 浓度以及计算出的 GSH/GSSG 摩尔比差异很大。在这里，我们详细描述了样品操作过程中发生的与 GSH 巯基 (SH) 基团人工氧化相关的主要分析和预分析问题。我们强调，自五十多年前首次描述以来，这一方面如何长期被忽视。最后，讨论了测量生物样品中 GSH 和 GSSG 的可靠程序和方法。
Glutathione (GSH) is the most abundant low-molecular-mass thiol within cells and one of the major antioxidant compounds in body fluids. Under pro-oxidant conditions, two GSH molecules donate one electron each and are converted into glutathione disulfide (GSSG). The GSH/GSSG molar ratio is considered a powerful index of oxidative stress and disease risk. Despite high interest in GSH/GSSG titration as measures of thiol redox balance, no broad agreement has yet been reached as to the best pre-analytical and analytical methods for the quantitation of these molecules in biological samples. Consequently, measured concentrations of GSH and GSSG and calculated GSH/GSSG molar ratios vary widely among laboratories. Here, we describe in detail the main analytical and pre-analytical problems related to the artificial oxidation of the sulfhydryl (SH) group of GSH that occur during sample manipulation. We underline how this aspect has been neglected for long time after its first description more than fifty years ago. Finally, selected reliable procedures and methods to measure GSH and GSSG in biological samples are discussed.