HPLC-MS法同时测定5种抗癫痫药的血药浓度

癫痫是神经系统的一种常见病,不少患者需长期同时服用两种或两种以上抗癫痫药物.本研究采用高效液相色谱-电喷雾离子化质谱法同时测定5种常用抗癫痫药物卡马西平、氯硝西泮、阿普唑仑、艾司唑仑和苯妥英钠的血药浓度,色谱条件为以地西泮作为内标,Zorbax SB-C18(30mm×2.1mm,3.5μm)为色谱柱,甲醇-25mM的乙酸铵水溶液梯度洗脱,样品采用无水乙醚萃取.样品分析以离子阱质谱作为检测器,采用多重反应监测模式(MRM),选择MS2离子提高方法的专属性,5种抗癫痫药物在50～5000ng/mL浓度范围内均具有良好的线性关系,其日内、日间精密度均小于10%,方法回收率均大于90%.研究表明,本方法专属性强,灵敏度高,线性关系良好,适用于这5种抗癫痫药物的血药浓度监测、个体化给药方案的制定、药物滥用监测以及药代动力学研究.关键词高效液相色谱质谱抗癫痫药血药浓度药物分析

Simultaneous determination of five anti-epilepsy drugs in human plasma using HPLC-MS

A new liquid chromatography-mass spectrometry method for the determination of carbamazepine,clonazepam,alprazolam,estazolam and phenytoin in human plasma has been developed by using diazepam as internal standard.Chromatographic separation was performed on a 3.5 μm Zorbax SB-C18 column（30 mm × 2.1 mm） with a mobile phase consisting of methanol and aqueous 25 mM ammonium acetate using gradient elution.A diethyl ether extraction method was used for the extraction of five anti-epilepsy drugs.Final extract was injected for analysis for LC-MS with MS2 mode.The method was validated within the concentration range of 50？5000 ng/mL for five anti-epilepsy drugs.The precision of the assay（RSD%） was less than 10% at all concentration levels within the tested range.The method recoveries for all samples were more than 90%.The results clearly indicate that the method is specific,sensitive and accurate,and suitable to study the pharmacokinetics,adjust the dosage for individual administration,and monitor the drug-concentration and drug abuse of the five anti-epilepsy drugs.