柱切换-高效液相色谱法快速监测癫痫患者卡马西平血药浓度

Rapid monitoring of carbamazepine blood drug level in epileptics by column- switching high performance liquid chromatography

目的:建立快速监测癫痫患者血清中卡马西平血药浓度的柱切换-高效液相色谱法。方法:通过直接进样方式,利用自制限进填料作为预处理柱,除去血清中蛋白质等大分子杂质,卡马西平保留在预处理柱上;再通过柱切换技术,使卡马西平在分析柱上得到分析检测。通过条件优化,确定切换前预处理流动相为甲醇-水(5∶95),流速为1.2 mL·min-1;切换后分析流动相为甲醇-水(70∶30),流速为1.0 mL·min-1;切换时间为3 min;柱温:25℃;检测波长:240 nm。结果:卡马西平在0.05～20μg·mL-1范围内具有良好的线性关系(r=0.9994),3个浓度水平(2、8、12μg·mL-1)的平均回收率分别为100.5%、100.8%、99.3%,日内和日间精密度均小于5%。结论:本方法快速、简便、灵敏、准确,适用于癫痫患者日常临床血药浓度监测。

Objective： To develop a high performance liquid chromatography（HPLC）method with column-switching for rapid monitoring of the blood drug level of carbamazepine in serum of epileptics. Methods： The column of restricted-access medium was used as the precolumn and an ODS was used as the analytical column.The epileptic serum samples were injected directly to remove plasma protein and endogenous plasma constituents as well as retain carbamazepine in the precolumn.Then carbamazepine was transferred by switching technology to the ODS with the mobile phase of methanol-water（70： 30,V/V）at the flow rate of 1.0 mL·min-1.The column-switching time was 3.0 min,the detection wavelength was 240 nm and the column temperature was set at 25℃. Results： The calibration curve showed excellent linear relationship（r=0.9994）in the concentration range of 0.05-20 μg·mL-1 for carbamazepine in epileptic serum.The intra-day and inter-day assay precisions and accuracies were good and the relative standard deviations（RSDS）were less than 5%.The average recoveries of carbamazepine at 3 concentrations（2,8,and 12 μg·mL-1）were 100.5%,100.8%,and 99.3%,respectively. Conclusion： The method is convenient,quick,sensitive,accurate,and suitable for the daily clinical monitoring of the blood drug level of carbamazepine in epileptics.