Simultaneous determination of 8-hydroxy-2′-deoxyguanosine and 5-methyl-2′-deoxycytidine in DNA sample by high performance liquid chromatography/positive electrospray ionization tandem mass spectrometry
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- 作者：Junjie Hu ; Wenbing Zhang ; Huimin Ma ; Yunmei Cai ; Guoying Sheng ; Jiamo Fu
- 关键词：8-Hydroxy-2′ ; -deoxyguanosine (8-OHdG) ; 5-Methyl-2′ ; -deoxycytidine (5-mdC) ; Electrospray tandem mass spectrometry ; Oxidative DNA damage ; DNA methylation
- 刊名：Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences
- 出版年：2010
- 出版时间：15 October 2010
- 年：2010
- 卷：878
- 期：28
- 页码：2765-2769
- 全文大小：397 K
文摘
8-Hydroxy-2′-deoxyguanosine (8-OHdG) and 5-methyl-2′-deoxycytidine (5-mdC) are utilized as useful biomarkers not only for early diagnosis but also for the detection and assessment of high-risk individuals. In the present study, a sensitive and specific method was developed for simultaneous determination of 8-OHdG and 5-mdC in DNA by high performance liquid chromatography/positive electrospray ionization tandem mass spectrometry. The limits of quantification for 8-OHdG and 5-mdC were 80 and 40 pg/ml, respectively. The calibration curves of 8-OHdG and 5-mdC were linear over the concentration range of 0.02–100 ng/ml and the correlation coefficients were higher than 0.9990. The intra-day and inter-day relative standard derivative values were in the range of 0.70–7.47 % for 8-OHdG and 1.07–7.06 % for 5-mdC, respectively. The recoveries were 93.4–108.5 % for 8-OHdG and 87.4–104.9 % for 5-mdC, respectively. This method was validated by determination of the background levels of 8-OHdG and 5-mdC in calf thymus DNA, and satisfactory results were obtained.