HPLC法同时测定人血清中卡马西平、环氧化卡马西平和苯妥英

HPLC Simultaneous Determination of Carbamazepine, Carbamazepine -10, 11 -epoxide, Phenytoin in Human Serum

目的:建立同时测定人血清中苯妥英、卡马西平及其代谢物10,11-环氧化卡马西平浓度的HPLC法。方法:用含内标氯唑沙宗的乙腈沉淀样品中蛋白,取上清液进样。分析柱为YWG-C_(18)(4.6mm×200mm,10μm),流动相为乙腈-甲醇-水(13:25:62),检测波长为214 nm。结果:卡马西平、10,11-环氧化卡马西平和苯妥英的标准曲线范围分别为0.4～16,0.2～8,1～40μg·mL^(-1),最低检测浓度分别为0.2,0.1,0.5μg·mL^(-1)。三者的方法回收率近100％,日内和日间精密度均小于6％,内源性物质和常用药物均不干扰测定。结论:该方法操作简便,灵敏、准确,适用于治疗药物监测。

Objective: To establish a HPLC method for the simultaneous determination of phenytoin ( PHT) , car-bamazepine (CBZ) and its metabolite (carbamazepine - 10, 11 - epoxide, CBZE) in human serum. Method: Acetonitrile, containing internal standard chlorzoxazone, was added to serum samples to precipitate the serum proteins. After centrifugation, the acetonitrile supernatant was injected for analysis by HPLC. Separation was a-chieved on a YWG - C18 analytical column (4.6 mm ×200 mm, 10μm) with a mobile phase of 13: 25: 62 acetonitrile - methanol -water. Detection was at 214 nm using an ultraviolet detector. Results: The calibration curve was linear within the range of 0. 4 - 16 μg·mL-1 of CBZ, 0. 2- 8 μg· mL-1 of CBZE and 1 -40 μg · mL-1 of PHT. The limit concentration of detection in CBZ, CBZE and PHT were 0.2, 0. 1 , 0. 5μg · mL-1 respectively. The average recovery of three drugs is nearly 100% and the RSD for the inter - and intra - day were less than 6% . Endogenous matrix and commonly used drugs did not interfere with the analysis. Conclusion: The method was convenient, sensitive and accurate. It was suitable for therapeutic monitoring of CBZ and/or PHT.