A simple high-performance liquid chromatographic assay for ciprofloxacin in human serum

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Abstract
A simple and selective high-performance liquid chromatographic (HPLC) method for the determination of ciprofloxacin in serum has been developed and evaluated. Serum protein was precipitated with acetonitrile. The drug and the internal standard (quinine) were eluated from a 10 μm U-Bondapack C-18 cartridge at ambient temperature with a mobile phase consisting of acetonitrile: 0.1 m sodium dihydrogen phosphate (20:80%, v/v) adjusted to pH 3.9 with phosphoric acid, and at a flow rate of 2.5 ml/min. The effluent was monitored on a fluorescence detector using an excitation and emission wavelength of 280 and 455 nm, respective. Each analysis required no longer than 6 min. Quantification was achieved by the measurement of the peak-height ratio and the limit of quantification for ciprofloxacin in serum is 25 ng/ml. The intraday coefficient of variation (CV) ranged from 0.4 to 5.8%, and interday CV from 4.6 to 8.8% at three different concentrations. Relative recovery ranged from 98 to 100.2% at three different concentrations. Preliminary stability tests show that ciprofloxacin is stable for at least 3 weeks in serum after freezing.