A robust ultra-performance liquid chromatography-tandem mass spectrometry method for simultaneous determination of 10 components in glutathione cycle

被引：0

作者：

Qin, Xiang-Ling ^{[1
]}

Wang, Si-Yu ^{[1
]}

Li, Qi-Long ^{[1
]}

Wang, Jun-Jie ^{[1
]}

Yao, Zhong-Wei ^{[1
]}

Zhu, Jin-Hao ^{[1
]}

Chen, Lin-Xia ^{[1
]}

Huo, Jie-Ge ^{[1
]}

Li, Song-Lin ^{[1
]}

Zhou, Jing ^{[1
]}

Zhu, He ^{[2
,3
]}

机构：

[1] Nanjing Univ Chinese Med, Affiliated Hosp Integrated Tradit Chinese & Wester, Dept Pharmaceut Anal, Nanjing 210028, Peoples R China

[2] Nanjing Med Univ, Affiliated Taizhou Peoples Hosp, Drug Clin Trial Ctr, Taizhou, Peoples R China

[3] Nanjing Univ Chinese Med, Affiliated Hosp Integrated Tradit Chinese & Wester, Nanjing, Peoples R China

来源：

JOURNAL OF SEPARATION SCIENCE | 2024年 / 47卷 / 12期

基金：

中国国家自然科学基金;

关键词：

glutathione; glutathione cycle; GSH; oxidative stress; UPLC-MS/MS; THIOL REDOX; VALIDATION;

D O I：

暂无

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

Glutathione (GSH) is an important antioxidant that is generated and degraded via the GSH cycle. Quantification of the main components in the GSH cycle is necessary to evaluate the process of GSH. In this study, a robust ultra-performance liquid chromatography-tandem mass spectrometry method for the simultaneous quantification of 10 components (GSH; gamma-glutamylcysteine; cysteinyl-glycine; n-acetylcysteine; homocysteine; cysteine; cystine; methionine; glutamate; pyroglutamic acid) in GSH cycle was developed. The approach was optimized in terms of derivative, chromatographic, and spectrometric conditions as well as sample preparation. The unstable thiol groups of GSH, gamma-glutamylcysteine, cysteinyl-glycine, n-acetylcysteine, cysteine, and homocysteine were derivatized by n-ethylmaleimide. The derivatized and underivatized analytes were separated on an amino column with gradient elution. The method was further validated in terms of selectivity (no interference), linearity (R-2 > 0.99), precision (% relative standard deviation [RSD%] range from 0.57 to 10.33), accuracy (% relative error [RE%] range from -3.42 to 10.92), stability (RSD% < 5.68, RE% range from -2.54 to 4.40), recovery (RSD% range from 1.87 to 7.87) and matrix effect (RSD% < 5.42). The validated method was applied to compare the components in the GSH cycle between normal and oxidative stress cells, which would be helpful in clarifying the effect of oxidative stress on the GSH cycle.

引用

页数：15

共 45 条

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