Simple, Rapid, and Sensitive Determination of Thiols by Liquid Chromatography with Fluorescence Detection
文献类型: 外文期刊
第一作者: Wang, Ya
作者: Wang, Ya;Yu, Xiangyang;Wang, Ya;Yu, Xiangyang;Wang, Ya;Zheng, Yanheng;Ge, Ying;Zhang, Chunhua
作者机构:
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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