Previous studies suggested that certain constituents of ambient PM2.5 can induce or increase oxidative stress in biological systems. The present study is designed to examine whether exposure to traffic generated particles increases the burden of oxidative stress in humans and to identify specific PM2.5 constituents responsible for pollution-induced oxidative stress. We recruited two nonsmoking security guards who worked at a university campus gate by a heavily trafficked road. Pre- and post-workshift spot urines were collected on each of the 29 days of measurement. Concentrations of PM2.5 mass and 126 chemical species were measured at the worksite and a campus background site simultaneously. Urine samples were analyzed for 8-hydroxy-2'-deoxyguanosine (8-OHdG). Factor analysis and linear mixed-effects regression models were used in statistical analyses. Three clusters of PM2.5 species were identified, including PAHs, metals, and polar organic compounds. Urinary concentrations of 8-OHdG increased by > 3 times following an eight-hour workshift in participants. Pre-workshift urinary concentrations of 8-OHdG were associated with PM2.5 concentrations at the background site. Post-workshift 8-OHdG concentrations were significantly and positively associated with PM2.5 mass, PAHs, and metals, but not polar organic species, measured at the worksite. Our findings provide direct evidence in humans that PM compositions are important in increasing oxidative stress burdens. Our results support that PAHs and metals are biologically active constituents of PM2.5 with regards to the induction of oxidative DNA damages in the human body.