title: [Analysis of 8-oxo-7,8-dihydro-2'-deoxyguanosine using ultra high performance liquid chromatography-tandem mass spectrometry].
abstract: An ultra high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the analysis of biomarker 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-OHdG) in deoxyribonucleic acid (DNA) was developed. The artificial oxidation of 2'-deoxyguanosine (dG) at numerous stages in the sample preparation bring a challenge to the accurate measurement of 8-OHdG in DNA. To avoid the artificial oxidation during the enzymatic digestion, desferrioxamine mesylate as a protectant was added into the mixtures. By utilizing YM-3 Centricon membrane (3 000 of relative molecular mass cut off), excess proteins in enzymatic solutions were effectively removed, allowing direct UPLC-MS/MS analysis. The UPLCMS/MS analysis showed a linear relationship between the peak areas and the amounts of 8-OHdG in the range of 17.6-1400 fmol, and the correlation coefficient was 0.9998. By using the developed method, the content of 8-OHdG in calf thymus DNA (CT DNA) was estimated about 12.9 +/- 2.35 (calculated as 8-OHdG/10(6) dG), which was consistent with the previous work. This method can also be applicable for the detection of 8-OHdG in DNA under various oxidative stresses.
authors: State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China.
authors: Yuling
authors: Y
authors: Song
authors: Hailin
authors: H
authors: Wang
mesh_terms: D000080242:8-Hydroxy-2'-Deoxyguanosine; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004247:DNA / Q000378:metabolism; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000032:analysis; D018384:Oxidative Stress; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013950:Thymus Gland / Q000378:metabolism
chemical_list: D015415:Biomarkers; D000080242:8-Hydroxy-2'-Deoxyguanosine; D004247:DNA; D003849:Deoxyguanosine