The metabolism and excretion of trovafloxacin was investigated in four healthy male volunteers after a single oral administration of 200 mg of [14C]trovafloxacin (118 microCi). Mean values of 23.1 and 63.3% of the administered dose were recovered in the urine and feces, respectively, after 240 hr. The Cmax of total radioactivity and unchanged trovafloxacin in serum was 3.2 micrograms-equiv/ml and 2.9 micrograms/ml, respectively, and peaked in 1.4 hr. The mean AUC0-infinity for radioactivity and trovafloxacin was 58.2 micrograms-eq.hr/ml and 32.2 micrograms.hr/ml, respectively. This implied that unchanged trovafloxacin constituted 55% of the circulating radioactivity. Urine and fecal samples were analyzed by LC/MS/MS for characterization of the metabolites, and the quantity of each metabolite in the matrices was assessed by means of a radioactivity detector. The profile of radioactivity in urine showed three main metabolites that were identified as the trovafloxacin glucuronide (M1), N-acetyltrovafloxacin glucuronide (M2), and N-acetyltrovafloxacin (M3). The major fecal metabolites were M3 and the sulfate conjugate of trovafloxacin (M4). Analysis of circulating metabolites from pooled serum extracts obtained at 1, 5, and 12 hr indicated that M1 was the major circulating metabolite (22% of circulating radioactivity), whereas M2 and M3 were detected in minor amounts. The results of the present study revealed that oxidative metabolism did not play a significant role in the elimination of trovafloxacin, and phase II conjugation was the primary route of trovafloxacin clearance in humans.