UPLC-MS/MS法测定Beagle犬血浆中多奈哌齐的质量浓度及其药动学研究

Determination of donepezil in Beagle dog plasma by UPLC-MS/MS and its application to pharmacokinetic study

目的建立测定Beagle犬血浆中多奈哌齐的UPLC-MS/MS方法,并将其应用于盐酸多奈哌齐片在Beagle犬体内的药物动力学研究。方法采用沉淀蛋白法进行样品预处理,色谱柱为Acquity UPLC BEH C_(18)(50 mm×2.1 mm,1.7μm)柱,流动相采用乙腈(含体积分数0.3%甲酸)-水(含体积分数0.3%甲酸),梯度洗脱,以正离子扫描多反应监测(MRM)模式进行检测。结果血浆中多奈哌齐在质量浓度0.10~100.80μg·L^(-1)内线性关系良好,日内和日间精密度RSD均不超过10%,提取回收率为83.9%~88.1%,基质效应为95.5%~100.1%,多奈哌齐血浆样品在所考察的储存条件下均可保持稳定。结论该方法适用于盐酸多奈哌齐片在Beagle犬体内的药物动力学研究,国内研制的盐酸多奈哌齐片和国外参比制剂在Beagle犬体内的药物动力学行为具有一致性。

Objective To establish an UPLC-MS/MS method for the determination of donepezil in dog plasma, and apply the method to the pharmacokinetic study of donepezil hydrochloride tablets in beagle dogs. Methods The plasma samples was pretreated by protein precipitation with acetonitrile, and chromatographic separation was performed on an Acquity UPLC BEH C18 column using a gradient elution with mobile phase consisting of acetonitrile （containing 0. 3 % formic acid）and 0. 3 % formic acid in water. The detection was carried out by multiple reactions monitoring in positive ion mode. Results The method was linear over the range of 0. 10 - 100. 80 μg· L ^-1 ,and the intra-and inter-day precision were below 10%. The extraction recovery ranged from 83.9% to 88.1%. The matrix effect was between 95.5% and 100. 1%. Donepezil plasma samples were found to be stable in the different conditions investigated in this article. Conclusions The method was successfully applied to the pharmacokinetic study of donepezil hydrochloride tablets in Beagle dogs, and the test formulations are bioequivalent to the reference formulations.