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.
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 40pg/ml, respectively. The calibration curves of 8-OHdG and 5-mdC were linear over the concentration range of 0.02-100ng/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.
Hu
Junjie
J
State Key Laboratory of Organic Geochemistry, Guangzhou Institute of Geochemistry, Chinese Academy of Sciences, Guangzhou 510640, China; Graduate school of the Chinese Academy of Science, Beijing, China.
Zhang
Wenbing
W
Ma
Huimin
H
Cai
Yunmei
Y
Sheng
Guoying
G
Fu
Jiamo
J
D000080242:8-Hydroxy-2'-Deoxyguanosine; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004247:DNA / Q000737:chemistry*; D004249:DNA Damage; D019175:DNA Methylation; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000032:analysis; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000032:analysis; D016014:Linear Models; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013950:Thymus Gland
D015415:Biomarkers; D003841:Deoxycytidine; D000080242:8-Hydroxy-2'-Deoxyguanosine; D004247:DNA; C016569:5-methyldeoxycytidine; D003849:Deoxyguanosine