Development of a targeted method for DNA adductome and its application as sensitive biomarkers of ambient air pollution exposure

DNA adducts are widely recognized as biomarkers of exposure to environmental carcinogens and associated health effects in toxicological and epidemiological studies. This study presents a targeted and sensitive method for comprehensive DNA adductome analysis using ultra-high-performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (UHPLC-QqQ-MS/MS). The method was developed using calf thymus DNA, with careful optimization of mass spectrometric parameters, chromatographic separation conditions, and pretreatment methods. Ultimately, a targeted method was established for 41 DNA adducts, which showed good linearity (R2≥0.992), recovery (80.1%–119.4%), accuracy (81.3%–117.8%), and precision (relative standard deviation <14.2%). The established method was employed to analyze DNA adducts in peripheral blood cells from pregnant women in Shanxi and Beijing. Up to 23 DNA adducts were successfully detected in samples of varying sizes. From 2 μg of maternal DNA samples, seven specific adducts were identified: 5-methyl-2'-deoxycytidine (5-MedC), 5-hydroxymethyl-2'-deoxycytidine (5-HmdC), N6-methyl-2'-deoxyadenosine (N6-MedA), 8-hydroxy-2'-deoxyguanosine (8-OHdG), 5-hydroxy-2'-deoxycytidine (5-OHdC), 1,N6-etheno-2'-deoxyadenosine (1,N6-εdA), and N2-methyl-2'-deoxyguanosine (N2-MedG). This study reveals that exposure to higher concentrations of ambient air pollutants may elevate the levels of DNA methylation and oxidative damage at different base sites, highlighting the application potential of DNA adducts as sensitive biomarkers of air pollution exposure. DNA adducts generated from environmental exposure can induce mutations, potentially resulting in adverse health outcomes. Quantitative analysis of DNA adducts is instrumental in evaluating the genotoxicity and potential pathogenicity of environmental pollutants. The development of a targeted DNA adductome method contributes significantly to revealing different types of DNA damage induced by environmental exposure and represents a novel approach to assessing environmental health risks. This method has been successfully applied to pregnant women, demonstrating potential clinical applicability by accurately identifying patterns of DNA damage and serving as sensitive biomarkers of ambient air pollution exposure.'