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Robust method for the analysis of phytochelatins in rice by high‐performance liquid chromatography coupled with electrospray tandem mass spectrometry based on polymeric column materials
Author(s) -
Yu Shasha,
Bian Yingfang,
Zhou Rong,
Mou Renxiang,
Chen Mingxue,
Cao Zhaoyun
Publication year - 2015
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201500557
Subject(s) - chemistry , chromatography , formic acid , tandem mass spectrometry , detection limit , phytochelatin , cadmium , mass spectrometry , electrospray , solid phase extraction , liquid chromatography–mass spectrometry , extraction (chemistry) , sample preparation , analytical chemistry (journal) , glutathione , biochemistry , organic chemistry , enzyme
A sensitive and robust high‐performance liquid chromatography coupled with electrospray tandem mass spectrometry method for the identification and quantification of glutathione and phytochelatins from rice was developed. Homogenized samples were extracted with water containing 100 mM dithiothreitol, and solid‐phase extraction using polymer anion exchange resin was employed for sample purification. Chromatography was performed on a polymeric column with acetonitrile and water containing 0.1% formic acid as the mobile phase at the flow rate of 300 μL/min. The limit of quantitation was 6–100 nM. This assay showed excellent linearity for both glutathione and phytochelatins over physiological normal ranges, with correlation coefficients ( r ) > 0.9976. Recoveries for four biothiols were within the range of 76–118%, within relative standard deviations less than 15%. The intraday precision ( n = 7) was 2.1–13.3%, and the interday precision over 15 days was 4.3–15.2%. The optimized method was applied to analyze tissue samples from rice grown using nutrient solutions with three different cadmium concentrations (0, 50, and 100 μM). With increasing cadmium concentrations, the content of phytochelatin 2 and phytochelatin 3 in rice roots increased, in contrast to most phytochelatins, and the content of glutathione in rice stems and roots decreased significantly.