摘要：Aim To develop a sensitive and accurate HPLC method for the determination of fleroxacin in human plasma, and study its pharmacokinetics in healthy subjects. Methods The analytes were isolated fi＇om plasma by simple protein precipitation with methanol, separated on a Diamonsil C18 column by isocratic elution with the mobile phase consisted of 1% triethylamine at pH 4.8 （adjusted with phosphoric acid） and acetonitrile （80/20, V/V） at a flow rate of 1.0 ***^-1 and analyzed by fluorescence detector with an excitation at 290 nm and emission 458 nm. The pharmacokinetic study of fleroxacin was performed according to a double period crossover design. Results The weighted （1/x） calibration curve was linear over the plasma concentration range of 0.025 - 8.00 μ***^-1 The inter- and intra-day precisions （RSD/%） were no more than 5.16%, and the method accuracies and extraction recoveries at three concentrations ranged firom 99.1% to 100.9%, and 86.7% to 92.0%, respectively. Following oral administration at a dose of 400 mg fleroxacin, the main pharmacokinetic parameters for test and reference capsules were Cmax5.08 ± 0.78 and 5.38 ± 1.40 μ***^-1, tmax 1.72 ±0.79 and 1.82 ± 0.78 h, t1/2 11.68 ± 1.27 and 11.38 ± 1.51 h^-1, AUC0-∞ 78.44 ± 11.44 and 76.53 ± 13.24 μ***^-1.h, respectively. Conclusion The method is sensitive and accurate, and suitable for human pharmacokinetic study of fleroxacin.