High performance liquid chromatographic analysis of cephalexin in serum and urine

A rapid, highly sensitive high performance liquid chromatographic method has been developed for the determination of cephalexin in serum and urine. Serum protein was precipitated with 1% zinc sulphate solution containing cephradine as the internal standard. The drugs were eluted from a 5 micron, C-18 reversed-phase column at ambient temperature with a mobile phase consisting of acetonitrile-methanol-acetate buffer of pH 4.2 (10:10:80%), at a flow rate of 1.4 ml/min with ultraviolet detection at 254 nm. Each analysis lasted 9 min. Quantification was achieved by the measurement of the peak-height ratio and the relative and absolute recoveries varied from 98 to 103%. Detection limits for cephalexin were 1 microgram/ml in serum and 5 micrograms/ml in urine. Within-run coefficient of variation ranged from 0.73 to 5.63% at three different concentrations for the serum and urine assay.