Nagarajan Janaki Sankarachari Krishnan, Vimal Chandran Sathish, George Renjith, Dubala Anil
Department of Pharmaceutical Analysis, J.S.S. College of Pharmacy, Udhagamandalam, Tamilnadu, India.
J Pharm Anal. 2013 Aug;3(4):285-291. doi: 10.1016/j.jpha.2013.02.003. Epub 2013 Feb 26.
A simple, rapid and selective liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry (LC-APCI-MS) assay method has been developed and fully validated for the simultaneous quantification of cefadroxil (CF) and clavulanic acid (CA) in human plasma. Analytes and internal standard (IS) were extracted from human plasma by solid-phase extraction (SPE) technique using Sam prep (3 mL, 100 mg) extraction cartridge. The extracted samples were chromatographed on a reverse phase C column using a mixture of methanol: acetonitrile: 2 mM ammonium acetate (pH 3.5) (25:25:50, v/v/v) as the mobile phase at a flow rate of 0.8 mL/min. Quantification of the analytes and IS were carried out using single quadrupole LC-APCI-MS through selected-ion monitoring (SIM) at 362 and 198, for CF and CA, respectively. Method validation was performed as per the FDA guidelines and the results met the acceptance criteria. Plasma concentration of CF and CA followed by the oral administration of CF/CA (500/125 mg) pill to healthy male volunteers (=12) was measured. Area under plasma concentration-time curve from 0 to 12 h (AUC ) and 0 h extrapolated to infinity (AUC) were calculated. The ratio of AUC /AUC was found to be >85% for all the subjects, as recommended by the FDA guidelines.
已开发出一种简单、快速且选择性的液相色谱 - 大气压化学电离 - 质谱（LC - APCI - MS）分析方法，并对其进行了全面验证，用于同时定量测定人血浆中的头孢羟氨苄（CF）和克拉维酸（CA）。使用Sam prep（3 mL，100 mg）萃取柱，通过固相萃取（SPE）技术从人血浆中提取分析物和内标（IS）。提取的样品在反相C柱上进行色谱分析，以甲醇：乙腈：2 mM醋酸铵（pH 3.5）（25:25:50，v/v/v）的混合物作为流动相，流速为0.8 mL/min。通过单四极杆LC - APCI - MS在m/z 362和198处分别对CF和CA进行选择离子监测（SIM）来定量分析物和IS。按照FDA指南进行方法验证，结果符合验收标准。对12名健康男性志愿者口服CF/CA（500/125 mg）药丸后，测定了CF和CA的血浆浓度。计算了0至12小时的血浆浓度 - 时间曲线下面积（AUC）以及外推至无穷大的0小时的AUC（AUC）。如FDA指南所建议，所有受试者的AUC/AUC比值均大于85%。
