In the study presented here, levofloxacin concentrations in serum samples and the aqueous humour (AH) of 16 patients undergoing cataract extraction were measured in order to determine the penetration characteristics of levofloxacin into the AH of the non-inflamed human eye. Cataract removal was performed at various times (from 90 to 270 min) after the end of a 30-min intravenous infusion of 500 mg of levofloxacin. Serum samples were obtained 1 h after the end of levofloxacin administration (Cmax); AH and a second serum sample were taken simultaneously during the operation, and the concentrations of levofloxacin in AH (C(AH)) and serum (C(S)) were determined using a rapid high-performance liquid chromatography assay. The mean Cmax was 6.07 microg/ml (range 3.75-9.53 microg/ml, SD 1.83). The mean C(AH) at the first hour following levofloxacin administration was 1.37 microg/ml (range 1.17-1.6 microg/ml, SD 0.22) and the mean ratio (R=C(AH)/C(S)) was 0.26 (range 0.24-0.3, SD 0.02). The mean C(AH) at 125-270 min following levofloxacin administration was 1.39 microg/ml (range 0.82-1.98 microg/ml, SD 0.33) and the mean R was 0.3 (range 0.15-0.53, SD 0.11). Of 16 patients, 15 had a C(AH) of >1 microg/ml 1 h after levofloxacin administration. In conclusion, 1 h after administration of 500 mg of levofloxacin, the levels obtained were higher than the MIC at which 90% of methicillin-susceptible Staphylococcus aureus and certain gram-negative bacteria strains are inhibited.