HPLC-MS/MS法测定人血浆中多奈哌齐及其代谢物6-O-去甲基多奈哌齐浓度

Determination of donepoezil and its active metabolite 6-O-desmethyl donepezil in human plasma by HPLC-MS/MS method

目的:建立HPLC-MS/MS法测定血浆中多奈哌齐及其代谢物6-O-去甲基多奈哌齐浓度。方法:采用HPLC-MS/MS法,加入内标后,样品用二氯甲烷为萃取剂提取后进样,以Shisheido C18(100 mm×4.6mm,5μm)为色谱柱,甲醇-5 mmol·L-1醋酸铵-甲酸(80∶20∶0.05)为流动相,流速为0.2 mL·min-1,柱温为室温。离子化方式为电喷雾离子化,多反应离子监测。结果:多奈哌齐及6-O-去甲基多奈哌齐标准曲线线性范围分别为0.05～15 ng·mL-1和0.008～2.4 ng·mL-1,定量下限分别为0.05和0.008 ng·mL-1,二者日内RSD及日间RSD均<9%,方法回收率在96%～103%之间。稳定性考察试验,二者血药浓度变化率均在±15%内。结论:该方法简单,快速,可满足多奈哌齐及6-O-去甲基多奈哌齐血药浓度的测定及多奈哌齐药代动力学研究。

Objective ：To develop a sensitive HPLC-MS/MS method for analyzing donepoezil and its active metabolite 6-O-desmethyl donepezil in human plasma, nethods：Donepoezil and 6-O-desmethyl donepezil in plas- ma samples were extracted with dichloromethane after addition of internal standard. The analytes went through Shis- heido C18 column （100 mm ×4.6 mm,5 μm） with mobile phase methanol-（5 mmol.L^-1） ammonium acetate-formic acid （80： 20：0.05 ） ;the flow rate was 0.2 mL.min^-1 ;the column temperature was ambient temperature ; ESI ＋ was performed with MRM mode. Results：The calibration curves of donepoezil and 6-O-desmethyl donepezil were linear in the ranges of 0.05 - 15 ng-mL^-1 and 0. 008 -2.4 ng.mL^-1. The LLOQs of donepoezil and 6-O-desmethyl done- pezil in plasma were 0.05 and 0. 008 ng. mL^-1, respectively. Both the intra- and inter-day RSDs were less than 9% and the relative recovery was 96% - 103%. The concentration change of the samples in stability assay was within ± 15%. Conclusion：The method is simple and sensitive with good recovery and can be used in analyzing donepoezil and 6-O-desmethyl donepezil in plasma samples.