The effect of prenatal exposure to polycyclic aromatic hydrocarbons (PAHs) at the environmental level on birth size was examined. The concentration of 1-hydroxypyrene (1-OHP) in spot urine samples collected from 50 pregnant women in Tokyo was measured and was related to birth weight, length, and head circumference of the newborns the subjects delivered. Analysis of inter- and intra-individual variation in 1-OHP concentration in spot urine samples from non-smoking subjects revealed that single spot urine can represent the subject's exposure to PAHs over several weeks and utility of spot urine for assessing PAHs exposure level of non-smokers was indicated. Geometric mean urinary concentration of 1-OHP of the subjects was 0.056 μmol/mol creatinine, and smoking mothers (n = 7) excreted more 1-OHP (0.200 μmol/mol-cre) than non-smoking mothers (0.046 μmol/mol-cre). Multiple regression analysis using birth outcomes as the dependent variable and urinary 1-OHP and other covariates as independent variables revealed that urinary 1-OHP was not a significant variable to explain birth outcomes among non-smoking mothers.