高效液相色谱-质谱联用法同时测定苯巴比妥、丙戊酸钠、苯妥英钠和卡马西平的血药浓度

Determination the concentrations of phenobarbital,sodium valproate,phenytoin sodium and carbamazepine in human plasma for simultaneous quatification by HPLC-MS

目的建立高效液相色谱-质谱联用法以便同时测定人血浆中苯巴比妥、丙戊酸钠、苯妥英钠和卡马西平（均为抗癫痫药物）的血药浓度。方法以霉酚酸为内标,血浆样品经乙腈直接沉淀蛋白后,采用Zorbax Eclipse XDB-C18色谱柱（2.1 mm×150.0 mm,3.5μm）进行分离,以0.05%乙酸铵-甲醇（47∶53）为流动相,流量0.2 mL.min-1,柱温30℃;通过电喷雾离子源（ESI）,正负离子同时检测。结果苯巴比妥、丙戊酸钠、苯妥英钠和卡马西平线性范围分别为0.5~80.0,2.5~200.0,0.5~40.0和0.2~24.0μg.mL-1;最低检测限分别为0.05,0.20,0.10和0.01μg.mL-1;γ均大于0.99。高、中、低3个浓度的日内精密度小于3%,日间精密度小于15%,绝对回收率均大于70%。结论本方法简便、快速、灵敏、重现性好,能够有效地检测人血浆中4种药物的含量。

Objective To establish a liquid chromatography-mass spectrometry（HPLC-MS） method for determination the concentration of phenobarbital（PB）,sodium valproate（VPA）,phenytoin sodium（DPH） and carbamazepine（CBZ） in human plasma.Methods Mycophenolic acid was used as internal standard.After one step protein precipitation,the analytes were chromatographed on a XDB-C18（2.1 mm×150.0 mm,3.5μm）.An isocratic mobile phase consisting of 0.05% ammonium acetate in water methanol（47∶53）,flow velocity is 0.2 mL·min-1,column temperature is 30 ℃.ESI electrospray ionization source in both negative and positive modes.Results The calibration curves of each drug showed a good linearity in a range of concentration between 0.5-80.0 μg·mL-1for phenobarbital,2.5-200.0 μg·mL-1 for sodium valproate,0.5-40.0 μg·mL-1for phenytoin sodium,and 0.2-24.0 μg·mL-1 for carbamazepine,all γ0.99.Intra and inter precision（RSD%） were less than 3% and 15%,respectively.The recoveries were above 70%.Conclusion The method was suitable to determine these four drugs in human plasma.