Micromethod for determination of ceftriaxone in plasma and urine by high-performance liquid chromatography

A sensitive and rapid high-performance liquid chromatographic method for the determination of ceftriaxone in human plasma and urine is described. A C18 reversed phase column is used; the mobile phase comprises water-methanol-triethylamine (750:250:4v/v/v) adjusted to pH 3 with orthophosphoric acid. Quantitation is performed at 270 nm with cefazolin as the internal standard. This method involves precipitation of proteins from fluids with acetonitrile followed by extraction of endogenous compounds with chloroform and injection of the upper aqueous phase on to the chromatograph. Relative standard deviations for between-day and within-day assays are 6.2%. The detection limit is 0.5 microg(-1) in plasma and urine. Studies of drug stability during sample storage, sample pretreatment and chromatography showed no degradation of ceftriaxone or of the internal standard. The method is convenient for clinical monitoring and for pharmacokinetic studies.