Sitafloxacin is a new antibiotic drug belonging to the fourth generation quinolone antibiotics. The aim of this study was to evaluate the pharmacokinetics (PK), safety profiles, and bioequivalence of test and reference 50-mg sitafloxacin tablets under fasting and fed conditions. The PK parameters, which were calculated with noncompartmental model, including maximum concentration, area under the concentration-time curve (AUC) from time 0 to the last quantifiable concentration, and AUC from time 0 to infinity. The bioequivalence was assessed on the basis of whether the 90% confidence intervals of the geometric mean ratio for the test/reference drugs fell within the accepted range of 80%-125%. Adverse events were monitored to assess safety. Finally, 80 healthy Chinese participants were enrolled, of which 40 were enrolled in the fasting study and the other 40 enrolled in the fed study. There was 1 participant in the fed trial who withdrew from the study because of failure to finish the high-fat meal. The geometric mean ratio and its 90% confidence interval for the maximum concentration, AUC from time 0 to the last quantifiable concentration, AUC from time 0 to infinity between the branded and generic sitafloxacin, under both fasting and fed conditions, were compliant with the predefined bioequivalence criteria of 80%-125%. No serious adverse events were observed in this study. Therefore, the findings indicate that generic and original sitafloxacin tablets have comparable bioequivalence and safety profiles.