This study was designed to measure the concentration of ofloxacin in aqueous humor after topical, oral and intravenous administration in 50 patients undergoing cataract extraction. In Group 1, ofloxacin 0.3% eyedrops were topically instilled ten times and the aqueous humor concentration was 2.73 +/- 0.88 microg/ml. In Group 2, ofloxacin 0.3% eyedrops were topically instilled six times and the aqueous humor concentration was 0.84 +/- 0.61 microg/ml. Aqueous humor concentration 12 hours after 200 mg oral dose in Group 3, was 0.38 +/- 0.12 microg/ml. In Group 4, patients were given ofloxacin as a single intravenous 200 mg dose and the aqueous humor concentration 2 hours after the end of infusion was 0.45 +/- 0.11 microg/ml. Concentrations were determined by high performance liquid chromatography (HPLC) with fluorescence detection. There was a significant difference between Group 1 and the other groups, but not between Group 2 and Groups 3, 4. It was concluded that ofloxacin penetrates the corneal and the blood-aqueous barriers and can achieve good aqueous levels when given topically and systematically. Ofloxacin can be applied topically for external bacterial infections such as conjunctivitis and keratitis. Systematically administered ofloxacin reached higher levels than the MIC for some bacteria which cause endophthalmitis.