用HPLC法快速测定多种抗癫癎药物的血药浓度

A rapid HPLC method for simultaneous determination of multiple antiepileptic drugs in human serum

目的：建立用HPLC法同时快速测定多种抗癫痫药物血药浓度的方法。方法：血样处理采用硫酸锌沉淀法，色谱分析采用Diamonsil C18色谱柱（250mm×4．6mm，5μm），Waters2695型液相色谱分析系统，流动相为甲醇-水（65：35），紫外检测波长225nm。实验数据用Waters Empower液相色谱工作站处理，以阿普唑仑为内标，通过峰面积进行定量。结果：苯巴比妥在2．5～80μg／mL（r=0．9995）、苯妥英钠在1．25～40μg／mL（r=0．9985）、卡马西平在0．625～20μg／mL（r=0．9998）、氯硝西泮在0．1～3．2μg／mL（r=0．9997）、艾司唑仑在0．125～4μg／mL（r=0．9994）、地西泮在0．125～4μg／mL（r=0．9987）间线性关系良好，各药的平均回收率及血浆最低检测浓度（S／N≥3）分别为苯巴比妥101．07％和0．250μg／mL。苯妥英钠100．80％和0．312μg／mL，卡马西平100．32％和0．025μg／mL，氯硝西泮99．28％和0．040μg／mL。艾司唑仑99．25％和0．031μg／mL，地西泮101．05％和0．062μg／mL。各药物低、中、高3种浓度的日内、日间RSD均≤10％。结论：此法采血量少、预处理快速简便、方法灵敏度和准确性高，适合临床常规血药浓度监测。

Objective： To establish a rapid HPLC method for simultaneous determination of plasma concentrations of multiple antiepileptic drugs. Methods： The blood samples were pretreated by simple sedimentation with 5% ZnSO4 , and then analysed by Waters 2965 HPI,C system on a Diamonsil C18 column（250 mm×4.6 mm, 5 μm） using methanol ： H2O （65 ： 35） as mobile phase. Quantification was achieved with UV detection at 225 nm based on peak area with alprazolam used as internal standard. Results： The method was found to be linear over the range of 2.5-80 μg/mL （r=0. 999 5） for phenobarbital, 1.25- 40 μg/mL （r= 0. 998 5） for phenytoin sodium, 0. 625-20 μg/mL （r= 0, 999 8） for carbamazepine, 0. 1-3. 2 μg/mL （r= 0. 999 7） for clonazepam, 0. 125-4 μg/mL（r=0. 999 4） for estazolam and 0. 125-4μg/mL （r= 0. 998 7） for diazepam. The mean recovery and minimal detectable concentrations （S/N≥3） were 101.07 % and 0. 250 μg/mL for phenobarbital, 100.80 % and 0. 312 μg/mL for phenytoin sodium, 100.32% and 0. 025 μg/mL for carbamazepine, 99.28% and 0. 040μg/mL for clonazepam, 99.25% and 0. 031 μg/mL for estazolam, 101.05% and 0. 062 μg/mL for diazepam, respectively. At three different concentrations, the intra- and inter-day precisions （RSD） were all below 10 %. Conclusion： The method appeared to be a rapid, convenient and accurate method for determining the plasma concentrations of antiepileptic drugs.