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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

作者: Wang, Xiaoli 1 ; Guo, Tao 2 ; Wang, Shanshan 1 ; Chen, Xiangfeng 1 ; Chen, Yue 1 ; Yuan, Jinpeng 1 ; Zhao, Rusong 1 ;

作者机构: 1.Shandong Acad Sci, Anal & Test Ctr, Key Lab Appl Technol Sophisticated Analyt Instrum, Jinan 250014, Shandong, Peoples R China

2.Shandong Acad Agr Sci, Shandong Rice Res Inst, Jinan, Shandong, Peoples R China

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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