Simultaneous determination of cefepime and grepafloxacin in human urine by high-performance liquid chromatography

A liquid chromatographic method with UV detection for simultaneous determination of cefepime and grepafloxacin has been developed. The method uses a C 18 column, equipped with a pre-column of the same material, and acetonitrile–0.1 M phosphoric acid/sodium hydroxide buffer (pH 3.0)–0.01 M n-octylamine (pH 3.0) as mobile phase in gradient mode. Mobile flow rate and sample volume injected were 1.3 mL min −1 and 20 μL, respectively. Detection wavelengths were 259 nm for cefepime and 278 nm for grepafloxacin. The retention times were 4.03 min for cefepime and 8.85 min for grepafloxacin, with detection limits of 1.0 and 1.1 μg mL −1, respectively. The method was applied to the determination of both antibiotics in spiked samples of human urine.