Determination of 5-hydroxymethyl-2′-deoxycytidine in Rice by High-performance Liquid Chromatography–Tandem Mass Spectrometry with Isotope Dilution. (13th October 2017)

• Record Type: Journal Article
• Authors: Wang, Xiaoli; Guo, Tao; Wang, Shanshan; Chen, Xiangfeng; Chen, Yue; Yuan, Jinpeng; Zhao, Rusong
• Is Part Of: Analytical letters. Volume 50:Number 15(2017)
• Page Start: 2351
• Page End: 2358
• Publication Date: 2017-10-13
• Subjects: 5-hydroxymethyl-2′-deoxycytidine -- high-performance liquid chromatography – tandem mass spectrometry (HPLC – tandem MS) -- HPLC-MS -- rice; Chemistry, Analytic -- Periodicals; Chemistry, Analytic -- Abstracts; 543
• DOI: 10.1080/00032719.2017.1286668
• Languages: English
• ISSNs: 0003-2719

Abstract

ABSTRACT: 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 5.0 pM. 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.