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Chem Res Toxicol. 2012 Feb 20;25(2):491-9. doi: 10.1021/tx200517h. Epub 2012 Jan 31.

Fast and simultaneous determination of urinary 8-hydroxy-2'-deoxyguanosine and ten monohydroxylated polycyclic aromatic hydrocarbons by liquid chromatography/tandem mass spectrometry.

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Key Laboratory of Ecology and Environmental Science in Guangdong Higher Education, College of Life Science, South China Normal University , Guangzhou, China 510631.


8-Hydroxy-2'-deoxyguanosine (8-OHdG), a biomarker of oxidative DNA damage, has been extensively studied to assess human exposure to carcinogenic compounds. Previous studies have associated levels of human urinary hydroxylated polycyclic aromatic hydrocarbons (OH-PAHs) with those of 8-OHdG. However, measurements of OH-PAHs and 8-OHdG in urine are often conducted with two different analytical methods, which is both costly and time-consuming. In this study, a novel method is described to quickly and simultaneously quantify ten urinary OH-PAHs and 8-OHdG through high pressure liquid chromatography/tandem mass spectrometry (HPLC/MS/MS). Urine samples undergo solid phase extraction and concentration and then are analyzed by an optimized HPLC/MS/MS method operated in the negative electrospray ionization (ESI) and multiple reaction monitoring (MRM) mode. Deuterated, (15)N and (13)C- labeled analogues are used as internal standards. Simultaneous analysis of urinary 8-OHdG and OH-PAHs are completed within 16 min. Calibration curves of all target analytes show favorable linearity within the concentration range of 0.3-10.0 μg/L for 8-OHdG and 0.05-15 μg/L for different OH-PAHs. The method detection limits (MDLs) in pooled urine range from 0.023 μg/L to 0.625 μg/L. The method shows satisfactory accuracy and precision when we analyzed varied levels spiked in pooled urine. Recoveries for 8 of the 10 OH-PAHs were in the range of 100 ± 15% with a variation coefficient of less than 20%. Thirty-four real urine samples were analyzed for all target analytes. Except 3-OHF, most compounds could be quantified.

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