Abstract: AIMTo establish a method for determineation of the concentration of ofloxacin in human fallopian tube, uterus and serum. METHODSThe separation was performed on a Spherisob C₁₈ column (Hypersil, 250 mm×4.6 mm ID, 5 μm) with a mobile phase of acetonitrile-0.01 moL·L^-1 potassium dihydrogen phosphate-0.5　mol·L^-1 tetrabutylammonium bromide (9∶91∶4, pH 2.5). The flow rate was 1.0 mL·min^-1 and detection was at 294 nm. The samples were homogenated or ground to powder after freezing with liquid nitrogen. 1%triton-100 and certain volume of ethylacetate-isopropanol (10∶1) were added, shaken and centrifuged. Then the entire organic layer was transferred to a tube and vacuum dried. The residue was reconstituted in the mobile phase for HPLC. RESULTSThere was a linear relationship between the peak area ratio and the ofloxacin concentration over the range of 0.2～8.0 μg·mL^-1. The limits of detection was 40 ng·mL^-1. Using this method to determine the ofloxacin concentrations in relevant organs as well as in the plasma of patients of the Department of Gynecology, and achieved satisfactary results. CONCLUSIONThe method can be applied to assay the ofloxacin concentration in human tissues. Ofloxacin was well distributed in woman fallopian tube, uterus and serum after single oral adminstration.