Determination of 5-hydroxymethyl-2-deoxycytidine in Rice by High-performance Liquid Chromatography-Tandem Mass Spectrometry with Isotope Dilution

文献类型： 外文期刊

第一作者： Wang, Xiaoli

作者： Wang, Xiaoli;Wang, Shanshan;Chen, Xiangfeng;Chen, Yue;Yuan, Jinpeng;Zhao, Rusong;Guo, Tao

作者机构：

关键词： 5-hydroxymethyl-2 '-deoxycytidine; high-performance liquid chromatography - tandem mass spectrometry (HPLC - tandem MS); HPLC-MS; rice

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： 2017 年 50 卷 15 期

页码：

收录情况： SCI

摘要： 5-hydroxymethyl-2-deoxycytidine (5-hmdC), which plays important roles in the epigenetic remodeling in mammals, has been recently determined in plant genomes at trace concentrations. In this study, a method for the sensitive, specific, and rapid assessment of 5-hmdC in rice is reported using high-performance liquid chromatography-tandem mass spectrometry (HPLC-tandem MS) with isotope dilution. The recovery of this method was from 75.6% to 81.3% for 5-hmdC and the intra- and inter-day precision values were within 5.81%. The quantification limit for 5-hmdC was 5.0pM. Genomic DNA from rice was extracted and enzymatically digested for nucleotides for the HPLC-tandem MS assay. The 5-hmdC levels were 1.32 +/- 0.08, 1.64 +/- 0.12, and 1.98 +/- 0.07 modifications per million nucleosides in the Shengdao-18, Shengxiang-145, and Jindao-263 rice cultivars. The application of HPLC-tandem MS for 5-hmdC demonstrated its presence in the genomic DNA of rice and is useful in further studying the role of modified cytosine in the epigenetic regulation of rice.

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