An investigation was made of the suitability of administering nicotine to experimental animals by inclusion in the drinking water. It was found that, after an initial accommodation period of several weeks, nicotine could be administered up to a concentration of 100 micrograms/ml with no decrease in fluid intake or weight gain compared to control. An analysis of the steady-state plasma levels and distribution of nicotine was made in mice which had received nicotine in the drinking water at a concentration of 60 micrograms/ml. The average daily dose of nicotine received by these animals was 17.2 mg/kg. The steady-state plasma level of nicotine was 34.4 ng/ml, representing 6% of the total compound present at steady-state as determined by thin-layer chromatography. The distribution of nicotine or metabolite in mice which had received [methyl-14C]-nicotine orally was determined. Whole-body autoradiography, as well as direct tissue counting, demonstrated that nicotine accumulates in a number of areas, particularly the salivary gland, nasal epithelium, uterus, and liver. There was relatively little material in the blood or brain. This investigation indicates that ad libitum oral administration is an acceptable method for maintaining experimental animals on nicotine for long periods of time.