LC-MS/MS法同时测定大鼠血浆中多奈哌齐和美金刚的含量

Simultaneous determination of donepezil and memantine in rat plasma by LC-MS/MS

目的建立一种简单、快速的LC-MS/MS法,同时测定大鼠血浆中多奈哌齐和美金刚的含量,研究多奈哌齐和美金刚在大鼠体内药动学的相互影响。方法大鼠血浆样品用正己烷-乙醚(体积比为1∶1)提取,Agilent-TC-C_(18)柱分离;流动相为甲醇-水(含体积分数为0.05%的甲酸溶液)(体积比为90∶10),流速为0.7m L·min^(-1),分流比为1∶1;质谱:采用电喷雾电离源,正离子方式检测,多反应监测扫描。结果多奈哌齐与美金刚线性范围分别为0.5~50.0μg·L^(-1)和2.0~200μg·L^(-1);定量下限分别为0.5μg·L^(-1)和2.0μg·L^(-1);准确度(RE)均小于15%,日内日间精密度(RSD)均不大于13.3%。结论该法简单、灵敏、选择性强,适用于同时测定大鼠血浆中多奈哌齐和美金刚的含量,并成功应用于多奈哌齐和美金刚在大鼠体内药动学相互影响的研究。

Objective To develop a simple and rapid LC-MS/MS method for simultaneous determination of donepezil and memantine in rat plasma. The interaction on pharmacokinetics in rat plasma of donepezil and memantine was investigated. Methods The plasma samples were extracted with hexane-diethyl ether （ V： V = 1： 1 ）, and the separation was performed on a reversed-phase Agilitent TC-C18 column. The mobile phase contained methanol and water with 0. 05% （φ） formic acid （ V： V = 90： 10）. The flow rate was 0.7 mL. min-1. Mass spectrometric detection was carried out by an electrospray ionization source, and the positive ion mode followed by multiple reaction monitoring （MRM）. Results The linear calibration curve of donepezil and memantine was obtained respectively in the range of 0. 5 - 50. 0 μg L-1 and 2. 0 - 200 μg. L-1. The lower limit of quantitation was 0. 5 μg L- 1 and 2.0 μg. L- 1, respectively. The accuracy （relative error ）was less than 15 %, and the intra-day and inter-day precision（RSD） was within 13.3%, and the recovery rate was more than 70%. Conclusions The method is simple, sensitive and selective. It is suitable for simultaneous determination of the concentrations of donepezil and memantine in rat plasma, and successfully applied to the interaction study of donepezil and memantine in rats.