正相HPLC-MS/MS测定人血浆中多奈哌齐对映体浓度

目的建立正相高效液相色谱-串联质谱(HPLC-MS/MS)法测定人血浆中多奈哌齐(DN)对映体的方法,并进行方法学验证。方法采用正己烷∶异丙醇(98∶2,V/V)作为萃取剂进行样品浓集,选用CHIRALCEL OJ-H手性柱(250mm×4.6mm,5μm)作分析柱,流动相为正己烷∶正丙醇∶二乙胺=60∶40∶0.1(V∶V∶V),流速为0.5mL/min,柱温为38℃。以多反应监测(MRM)方式进行定量检测,利多卡因为内标,按内标法定量。结果左旋DN(S-DN)、右旋DN(R-DN)对映体的保留时间分别为15.56和18.41min。S-DN的线性范围为0.051～7.596ng/mL,R-DN的线性范围为0.049～7.404ng/mL,线性良好(r>0.99)。S-DN和R-DN的方法回收率分别为95.10%～103.70%、93.58%～98.00%;预处理回收率分别为58.42%～61.08%、53.24%～61.87%;日内精密度(RSD)分别为8.35%～11.28%、6.78%～11.58%;日间RSD分别为5.82%～9.02%、6.87%～9.19%。结论建立的正相HPLC-MS/MS测定人血浆多奈哌齐对映体血药浓度具有快速、简便、灵敏、准确等优点,适用于多奈哌齐对映体的人体药动学研究。

Determination of Donepezil Enantiomer in Human Plasma by Normal-HPLC-MS/MS

Objective To develop and validate a normal phase HPLC-MS/MS method for the determination of donepezil enantiomer in human plasma.Methods Donepezil was extracted from plasma by n-hexane∶isopropanol(98∶2,V/V) with lidocaine serving as an internal standard.The analytes went through the column of CHIRALCEL OJ-H(250 mm×4.6 mm,5 μm) with mobile phase n-hexane∶n-propanol∶diethylamine(60∶40∶0.1,V/V/V).Donepezil enantiomer was determined by API 3000 in MRM mode.Results The retention time of S-DN and R-DN were 15.56 min and 18.41 min,respectively.The calibration curves were linear in a range from 0.051 to 7.596 ng/mL for S-DN,and from 0.049 to 7.404 ng/mL for R-DN,respectively,both with more than 0.99 correlation coefficients.The relative recovery were 95.10%-103.70% for S-DN and 93.58%-98.00% for R-DN,respectively;the pretreatment recovery were 58.42%-61.08% for S-DN and 53.24%-61.87% for R-DN,respectively;the within-day RSD ranged from 8.35% to 11.28% for S-DN and from 6.78% to 11.58% for R-DN,respectively;the between-day RSD ranged from 5.82% to 9.02% for S-DN and from 6.87% to 9.19% for R-DN,respectively.Conclusion This normal phase HPLC-MS/MS method is simple,rapid,sensitive and accurate for the determination of donepezil enantiomer in human plasma and is suitable for pharmacokinetic studies of donepezil enantiomers.