数字农科院2.0

A novel method for the simultaneous analysis of seven biothiols in rice (Oryza sativa L.) using hydrophilic interaction chromatography coupled with electrospray tandem mass spectrometry

文献类型： 外文期刊

作者： Cao, Zhao-Yun;Mou, Ren-Xiang;Zhou, Rong;Zhu, Zhi-Wei;Chen, Ming-Xue;Cao, Zhao-Yun;Mou, Ren-Xiang;Zhou, Rong;Zhu, Zhi-Wei;Chen, Ming-Xue;Sun, Li-Hua

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关键词： Biothiol quantification;Rice;Cd stress;Amide-HILIC;ESI-MS/MS

期刊名称： JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES

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收录情况： SCI

摘要： Analysis of biothiols is still problematic, due to their high polarity, oxidation sensitivity and time-consuming sample preparation. In this paper, a direct, rapid and sensitive method was developed for simultaneous quantification of unbound cysteine (Cys), glutathione (GSH) and phytochelatins (PCs) in rice leaf, stem and root samples by hydrophilic interaction chromatography coupled with electrospray tandem mass spectrometry (HILIC-MS/MS). Homogenized samples were extracted with water containing 50 mM dithiothreitol, without derivatization and further clean-up, and the extracts were injected directly onto an Xbridge Amide-HILIC column (3.5 mu m, 150 mm x 2.1 mm i.d.). The best chromatographic separation and MS sensitivity was achieved using a linear gradient elution with 10 mM aqueous ammonium formate and acetonitrile as the mobile phase. In MS/MS mode the detection limit (S/N >= 3) of seven biothiols was 3-105 nM. Good linearities were observed (r > 0.995) with linear dynamic range at least over three orders of magnitude. Recoveries for most analytes were within the range of 77-128%, with relative standard deviations less than 18.2%. The intra-day precision (n = 7) was 6.1-11.7%, and the inter-day precision over 15 d (n = 15) was 8.5-16.3% for all biothiols. The optimized HILIC-MS/MS method was applied to study the influence of different cadmium (Cd) concentrations (0, 1 and 50 mu M) on contents of Cys, GSH and PC2-6 in rice tissue. With increasing Cd concentrations in nutrient solutions, contents of PC2-4 in rice roots increased but contents of Cys and GSH decreased. Contents of PC2-4 in both rice leafs and stems increased markedly at high dose Cd (50 mu M) treatment compared with controls, compared with low Cd concentrations (1 mu M). However, both PC5 and PC6 were not detected throughout the stress experiment. (C) 2014 Elsevier B.V. All rights reserved.

分类号： Q503

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