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3003191 
Journal Article 
Ultrasensitive UPLC-MS/MS method for analysis of etheno-DNA adducts in human white blood cells 
Li, H; Cui, S; Wang, S; Jiang, X; Zhang, S; Zhang, R; Fu, PP; Sun, X 
2015 
Yes 
Free Radical Research
ISSN: 1071-5762
EISSN: 1029-2470 
49 
9 
1049-1054 
English 
25968941 
WOS:000360714300001 
Etheno-DNA adducts are generated by interaction of cellular DNA with exogenous environmental carcinogens and end products of lipid peroxidation. It has been determined that 1,N-6-etheno-2'-deoxyadenosine (epsilon dA) and 3,N-4-etheno-2'-deoxycytidine (epsilon dC) adducts formed in human white blood cells can be used to serve as biomarkers of genetic damage mediated by oxidative stress. In this study, we developed an ultrasensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method used to detect and quantify epsilon dA and epsilon dC adducts in human white blood cells. The percent recoveries of epsilon dA and epsilon dC adducts were found to be 88.9% + 2.8 and 95.7% +/- 3.7, respectively. The detection limits were similar to 1.45 fmol for epsilon dA and similar to 1.27 fmol for epsilon dC in 20 mu g of human white blood cell DNA samples, both epsilon dA and epsilon dC adducts could be detected using only similar to 5 mu g of DNA per sample. For validation of the method, 34 human blood cell DNA samples were assayed and the results revealed a significant difference (P < 0.01) between levels (fmol/mu g DNA) of 0.82 +/- 0.83 (standard deviation [SD]) (range: 0.15-3.11) for epsilon dA, 3.28 +/- 3.15 (SD) (range: 0.05-9.6) for epsilon dC in benzene-exposed workers; and 0.04 +/- 0.08 (SD) (range: 0.0-0.27) for epsilon dA and 0.77 +/- 1.02 (SD) (range: 0.10-4.11) for epsilon dC in non-benzene-exposed workers. Our method shows a high sensitivity and specificity when applied to small amounts of human white blood cell DNA samples; background levels of epsilon dA and epsilon dC could be reproducibly detected. The ultrasensitive and simple detection method is thus suitable for applications in human biomonitoring and molecular epidemiology studies. 
UPLC-MS/MS; 1, N-6-etheno-2'-deoxyadenosine; 3, N-4-etheno-2'-deoxycytidine; lipid peroxidation; human biomonitoring 
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