Abstract

8-Oxoguanosine (8-oxoGuo), 8-oxo-2′-deoxyguanosine (8-oxo-dG) and 8-iso-prostaglandin F2α (8-iso-PGF2α) are widely used noninvasive and effective biomarkers of oxidative damage to RNA, DNA and lipids. These three compounds are usually analyzed with different methods because of their different polarity and MS detection modes. In this paper, a sensitive and rapid UHPLC–MS/MS method to determine 8-oxoGuo, 8-oxo-dG and 8-iso-PGF2α in human urine was developed in order to evaluate the oxidative damage of DNA, RNA and lipid peroxidation. With this method, 8-oxoGuo, 8-oxo-dG and 8-iso-PGF2α could be conveniently and separately eluted with different elution solvent on the same Agilent Bond C18 solid-phase column; the two obtained eluates are detected through two injections using the same HPLC column and mobile phase for the determination of 8-oxoGuo, 8-oxo-dG and 8-iso-PGF2α in positive and negative ESI mode, respectively. The limits of detection and quantitation of the three biomarkers approached to 0.04–0.17 ng mL−1 and 0.13–0.57 ng mL−1, respectively. The intraday and interday precisions of all compounds were at most 10%. The method was applied to analyze urine samples from 40 smokers and 20 nonsmokers to evaluate the effect of cigarette smoking on oxidative stress. The results showed that concentrations of the three compounds in the urine of smokers are higher than those in nonsmokers, indicating the increased oxidative stress effect of smoking.

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