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Simple, Rapid, and Sensitive Determination of Thiols by Liquid Chromatography with Fluorescence Detection

文献类型： 外文期刊

作者： Wang, Ya 1 ; Zhang, Chunhua 4 ; Zheng, Yanheng 3 ; Ge, Ying 3 ; Yu, Xiangyang 1 ;

作者机构： 1.Minist Sci & Technol, State Key Lab Cultivat, Jiangsu Key Lab Food Qual & Safety, Nanjing 210014, Jiangsu, Peoples R China

2.Jiangsu Acad Agr Sci, Inst Food Safety & Nutr, Nanjing, Jiangsu, Peoples R China

3.Nanjing Agr Univ, Coll Resources & Environm Sci, Jiangsu Prov Key Lab Marine Biol, Nanjing 210095, Jiangsu, Peoples R China

4.Nanjing Agr Univ, Coll Life Sci, Lab Ctr Life Sci, Demonstrat Lab Element & Life Sci Res, Nanjing, Jiangsu, Peoples R China

关键词： High-performance liquid chromatography (HPLC); monobromobimane; thiol compounds; tris (2-carboxyethyl) phosphine hydrochloride

期刊名称：ANALYTICAL LETTERS （ 影响因子：2.329； 五年影响因子：1.738 ）

ISSN： 0003-2719

年卷期： 2019 年 52 卷 9 期

页码：

收录情况： SCI

摘要： Thiol compounds are important for protecting cells from oxidative stress. One common method of quantifying thiols is liquid chromatographic separation with fluorescence detection of their derivatives. The pH and the concentration of tris (2-carboxyethyl) phosphine hydrochloride in the reaction medium were shown to have significant effects on the fluorescence intensity of five thiol compounds: cysteine, glutathione, and three phytochelatins. The optimal pH range for derivatization, as indicated by the maximum fluorescence intensities, was 7.75-8.0 for all of the evaluated thiols. The thiol derivative fluorescence increased and then decreased with the tris (2-carboxyethyl) phosphine hydrochloride concentration. In particular, the fluorescence intensities of all of the derivatives decreased by 96.5-99.9% when tris (2-carboxyethyl) phosphine hydrochloride levels were increased from 0.1 to 1 mmol L-1. We attributed these changes to preferential interactions between tris (2-carboxyethyl) phosphine hydrochloride and the thiol-specific fluorophore, monobromobimane. We describe herein a method, based on our optimized solution pH and tris (2-carboxyethyl) phosphine hydrochloride concentration, that is rapid (12 min) and boasts excellent recovery (91.3-102%), sensitivity (limit of detections, 17.8-75.2 pmol L-1) and precision (relative standard deviation values <= 1.03%) for the quantification of these thiol compounds in microalgal samples.

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