Centro Cardiologico Monzino, IRCCS, Via Parea 4, 20138 Milan, Italy.
J Pharm Biomed Anal. 2012 Dec;71:111-8. doi: 10.1016/j.jpba.2012.08.013. Epub 2012 Aug 23.
Glutathione plays a central role in the defence against oxidative damage and in signaling pathways. Upon oxidation the reduced glutathione (GSH) is transformed to glutathione disulfide (GSSG). The concentration of GSH and GSSG in whole blood samples and their ratios is useful indicator of the oxidative stress status and/or disease risk. Here we describe a liquid-chromatographic method coupled with tandem mass spectrometry (LC-MS/MS) and we present the results of its comparison with a high-performance liquid-chromatographic method with electrochemical detection (HPLC-ECD). The method performed well in terms of validation parameters, i.e. linear range (0.01-50μM for both GSH and GSSG), precision (intra- and inter-day coefficients of variation were less than 10% for both GSH and GSSG), accuracy (bias% varied between -2.1 and 7.9% for both analytes), quantification limits (LLOQs were 0.5μM and 0.0625μM for GSH and GSSG respectively). Furthermore the LC-MS/MS method showed a good agreement with the HPLC-ECD assay. However, major benefits of LC-MS/MS are the improved selectivity, precision and accuracy, the higher sensitivity and the unaltered capacity of detection with time in contrast to ECD.
谷胱甘肽在抗氧化损伤和信号通路中起着核心作用。还原型谷胱甘肽 (GSH) 氧化后转化为谷胱甘肽二硫化物 (GSSG)。全血样本中 GSH 和 GSSG 的浓度及其比值是氧化应激状态和/或疾病风险的有用指标。在这里，我们描述了一种与串联质谱 (LC-MS/MS) 相结合的液相色谱方法，并介绍了与高效液相色谱-电化学检测 (HPLC-ECD) 方法进行比较的结果。该方法在验证参数方面表现良好，即线性范围 (GSH 和 GSSG 的线性范围均为 0.01-50μM)、精密度 (GSH 和 GSSG 的日内和日间变异系数均小于 10%)、准确度 (两种分析物的偏差%在-2.1%和 7.9%之间变化)、定量限 (GSH 和 GSSG 的定量限分别为 0.5μM 和 0.0625μM)。此外，LC-MS/MS 方法与 HPLC-ECD 测定法具有良好的一致性。然而，与 ECD 相比，LC-MS/MS 的主要优势在于提高了选择性、精密度和准确度、更高的灵敏度以及随时间推移检测能力不变。
