Determination of BMS-284756, a new quinolone, in mouse serum by high-performance liquid chromatography with fluorescence detection

A sensitive, simple, and accurate method for determination of BMS-284756, a novel des-F(6)-quinolone antimicrobial agent in mouse serum was developed by HPLC with fluorescence detection. Sample preparations were carried out by protein precipitation with the addition of acetonitrile, followed by evaporation of the acetonitrile to dryness. The resultant residual was then reconstituted in 0.01 M HCl and injected onto a Nucleosil 100 10 μm, C₁₈ 25 cm×4.6 mm analytical column. The mobile phase consisted of acetonitrile–0.01 M NaH₂PO₄ (20:80, v/v) with 0.01 M tetrabutylammonium hydrogen sulfate. The fluorescence of the column effluent was monitored at an excitation wavelength of 290 nm and an emission wavelength of 418 nm. The assay was shown to be linear from 0.2 to 10.0 μg/ml (R²=0.998). Mean recovery was determined as 95.1%. Inter- and intra-assay precisions were <6% RSD. The HPLC method developed has been applied to determine the pharmacokinetics of BMS-284756 in a murine bacterial infection model.