Simultaneous Determination of Cefuroxime and Sulbactam in Human Plasma and Urine by High Performance Liquid Chromatography
A HPLC-UV method was established for the simultaneous determination of Cefuroxime(CXM) and Sulbactam(SUL) in human plasma and urine. The chromatogram column was Welch Materials XB-C18(150 mm×4.6 mm, 5 μm). The mobile phase consisted of acetonitrile-0.01 mol/L potassium dihydrogen phosphate-triethylamine (15∶85∶0.04 for plasma, 10∶90∶0.04 for urine, pH 3.2), at a flow-rate of 1 mL/min. The detection wavelengths were set at 220 nm for SUL, and 274 nm for both CXM and the internal standard (caffeic acid). The acidific plasma sample was extracted twice with distilled diethyl ether and the urine sample was directly diluted with water. The calibration curves of SUL and CXM were linear range from 0.25 to 200 mg/L and 0.5 to 400 mg/L in plasma, respectively. The intra-day and inter-day RSD of SUL and CXM were both less than 9%, the method recoveries were 94.6%-113.6% and 91.9%-101.8%, the extraction recoveries were 75.6%-88.1% and 68.6%-77.4%, respectively. The calibration curves of SUL and CXM were linear in the range from 5 to 5120 mg/L and 10 to 10240 mg/L in urine, respectively. The intra-day and inter-day RSD of SUL and CXM was both than less 3%; the method recoveries were 92.9%-111.1% and 98.0%-105.3%, respectively. The method was found to be simple, rapid, sensitive and accurate for the determination of SUL and CXM in human plasma and urine. Besides, a HPLC-MS/MS was use to study the oxides of cefuroxime for first time.