Determination of the Active Metabolite of Prulifloxacin in Human Plasma by HPLC with Fluorescence Detection

A cheap, simple and rapid sample preparation method has been developed for quantification of ulifloxacin, the active metabolite of prulifloxacin in human plasma, by HPLC with fluorescence detection using lemefloxacin as the internal standard. One-step protein precipitation with 10% perchloric acid (2:1, v/v) on a 200 μL sample was used. The separation was performed at 30 °C on a C18 column using an eluent of acetonitrile-0.5% triethylamine buffer. The compounds were monitored at λex of 280 nm, λem of 425 nm. The calibration curve for ulifloxacin in human plasma was linear over the range 0.01–1.00 μg mL−1. The lower limit of quantification is 0.01 μg mL−1. The intra- and inter-day precision ranged from 3.0 to 6.7%, respectively. The method had been used for clinical pharmacokinetic studies of prulifloxacin formulation product after oral administration to healthy volunteers.