The validity of urinary nicotine and its metabolites as a biomarker of exposure to environmental tobacco smoke (ETS) has been investigated. After exposure to ETS, urine samples were collected from 10 subjects for the analyses of nicotine, cotinine and 3'-hydroxycotinine. The former two chemicals were detected in the urine of all subjects, and 3'-hydroxycotinine was detected in the urine of 9 subjects out of 10, indicating these three chemicals can be used as a biomarker of ETS exposure. 3'-Hydroxycotinine was not detected in the urine of one subject, suggesting that this subject may be a poor metabolizer of nicotine. In 9 subjects with 3'-hydroxycotinine excreted, the amounts of nicotine and cotinine started to increase after exposure, reached the peak at the end of the second exposure and decreased gradually. 3'-Hydroxycotinine started to be excreted into urine from 3 hours after exposure and kept the same level until 72 hours after exposure. In the urine of 72 hours after exposure, the amount of 3'-hydroxycotinine was the highest among these three chemicals.