Experiments with volunteers in Singapore have demonstrated that coffee drinking increases urinary hydrogen peroxide levels (Long, Halliwell, Free Rad. Res., 32, 463-467 (2000)). We re-examined the effect of coffee drinking of healthy Japanese subjects on urinary hydrogen peroxide levels. A cup of brewed or canned coffee commercially available in Japan generated 120-420 m mmol hydrogen peroxide in incubation in a neutral medium at 37°C for 6 h. The increased levels were higher than those obtained from a cup of green tea extract or a glass of red wine. After the subject drank a cup of coffee, apparent hydrogen peroxide levels (m mmol/g creatinine) in urine collected 1-2 h after coffee drinking increased 3-10-fold compared to the levels before coffee drinking. The increased urinary hydrogen peroxide levels are likely derived mainly from 1,2,4-benzenetriol excreted in urine, because the major component that generates hydrogen peroxide is found to be 1,2,4-benzenetriol, and storing urine collected after coffee drinking increased hydrogen peroxide levels in a time-dependent fashion. Total hydrogen peroxide equivalent levels excreted in 3 h-urine after coffee drinking were estimated to be 0.5-10% that of coffee consumed. A residual amount of hydrogen peroxide may be retained or consumed in human bodies.