Consumption of alcoholic beverages may suppress circulating melatonin levels at night, possibly resulting in an increase in circulating estrogen. An increased estrogen burden could increase the risk of breast cancer. This study was designed to investigate whether alcohol consumption is associated with a decrease in nighttime melatonin levels in a group of healthy women. A total of 203 randomly selected healthy women between the ages of 20 and 74 years were recruited for a broader study of the effects of exposure to power-frequency magnetic fields on nocturnal levels of urinary 6-sulfatoxymelatonin. For the purposes of this analysis, data collection consisted of the following during two seasons of the year: (1) an in-person interview, (2) a daily activity diary, and (3) nocturnal urine collection for each of 3 consecutive nights. We found that the nocturnal urinary concentration of the primary metabolite of melatonin (6-sulfatoxymelatonin) decreased in a dose-dependent manner with increasing consumption of alcoholic beverages in the preceding 24-hour period, after taking into account the independent effects on melatonin of age, hours of darkness, use of medications that affect melatonin levels, and body mass index. A categorical analysis revealed no effect of one drink, but a 9% reduction with two drinks, a 15% reduction with three drinks, and a 17% reduction with four or more drinks. It remains unknown whether such a change could affect estrogen levels or breast cancer risk.