Sensitive and simultaneous determination of 5-methylcytosine and its oxidation products in genomic DNA by chemical derivatization coupled with liquid chromatography-tandem mass spectrometry analysis.
作者： Yang Tang , Shu-Jian Zheng , Chu-Bo Qi , Yu-Qi Feng , Bi-Feng Yuan
DOI: 10.1021/AC504786R
关键词:
摘要: Cytosine methylation (5-methylcytosine, 5-mC) in genomic DNA is an important epigenetic mark that has regulatory roles diverse biological processes. 5-mC can be oxidized stepwise by the ten–eleven translocation (TET) proteins to form 5-hydroxymethylcytosine (5-hmC), 5-formylcytosine (5-foC), and 5-carboxylcytosine (5-caC), which constitutes active demethylation pathway mammals. Owing extremely limited contents of endogenous oxidation products, no reported method directly determine all these cytosine modifications simultaneously. In current study, we developed selective derivatization moieties with 2-bromo-1-(4-dimethylamino-phenyl)-ethanone (BDAPE) coupled liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) for simultaneous determination DNA. The chemical notably improved chromatography separation dramatically increased detection sensitivities of...
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