Household smoking is one of the main sources of environmental tobacco smoke (ETS) exposure for children, a population considered to be at high risk for associated negative health outcomes. Several studies evidenced the occurrence of early effects related to ETS exposure, including the development of the oxidative stress process. The aim of this study was to evaluate the correlation between urinary levels of 8-oxo-7,8-dihydro-2-deoxyguanosine (8oxodGuo), a nucleic acid oxidation biomarker, and socio-demographic features and lifestyle factors in school children (aged 5-11 years). A cross-sectional study was conducted among 154 healthy children, residing in rural zones of central Italy. For each participant, one urine sample was analyzed by the HPLC-MS/MS technique to simultaneously quantify 8oxodGuo and cotinine (a biomarker of ETS exposure), while information on the children was collected using a questionnaire filled out by the parents. Urinary levels of 8oxodGuo was found to be significantly higher in children exposed to ETS compared to those not exposed (5.53 vs. 4.78 μg/L; p = 0.019). This result was confirmed by the significant association observed between urinary levels of cotinine and 8oxodGuo (r = 0.364, p < 0.0001). Additionally, children exposed to ETS with no smoking ban at home showed a further increased difference than those not exposed (6.35 μg/L vs. 4.78 μg/L; p = 0.008). Considering the great number of adverse effects on human health due to exposure to passive smoking, especially if this exposure begins early in life, it is essential to implement health promotion interventions in this area.