超高效液相色谱-串联质谱法测定人血浆中非索非那定的浓度及其应用  

Determination of fexofenadine in human plasma by UHPLC-MS/MS and its application

作　　者：周玄 张燕 罗廷顺 曹昌娥 杨强丽 赖泳 ZHOU Xuan;ZHANG Yan;LUO Ting-shun;CAO Chang-e;YANG Qiang-li;LAI Yong(Col-lege of Pharmacy and Chemistry,Dali University,Yunnan Dali 671000,China;Department of Pharmacy,The First Affiliated Hospital,Dali University,Yunnan Dali 671000,China;Dali State Comprehensive Technical Inspection Center,Yunnan Dali 671000,China;Dali Institute for Food and Drug Control,Yunnan Dali 671000,China)

机构地区：[1]大理大学药学与化学学院,云南大理671000 [2]大理大学第一附属医院药剂科,云南大理671000 [3]大理州质量技术监督综合检测中心,云南大理671000 [4]大理州食品药品检验所,云南大理671000

出　　处：《中国医院药学杂志》2018年第16期1707-1711,共5页Chinese Journal of Hospital Pharmacy

基　　金：国家自然科学基金项目资助项目(编号:81360511)

摘　　要：目的:建立超高效液相色谱-串联质谱法(UHPLC-MS/MS)测定人血浆中非索非那定的浓度,并用于灯盏花素对人P-糖蛋白体内活性影响的研究。方法:200μL血浆经500μL乙腈沉淀蛋白后,采用Waters ACQUITY UPLC?BEH C18柱(50 mm×2.1 mm,1.7μm)为色谱柱,以乙腈(A)-0.1%甲酸水(B)为流动相梯度洗脱,流速为0.3 mL·min-1;质谱采用电喷雾正离子模式离子化(ESI+)和多反应监测模式(MRM)扫描,非索非那定和苯海拉明(内标)的定量离子对分别为m/z502.2→466.5和m/z256.1→167.1。结果:非索非那定和苯海拉明的保留时间分别为2.19 min和1.89 min。人血浆中非索非那定的提取回收率、方法过程效率和基质效应均值分别为84.3%~89.6%、86.3%~90.8%和101.2%~104.2%;线性范围为1.00~1 000 ng·mL-1(r=0.999 5);日内、日间精密度RSD≤10.9%,准确度RE%为-7.6%~5.0%;稳定性RSD和RE均在±15%内。18名志愿者给予灯盏花素或安慰剂后,非索非那定的达峰浓度Cmax为(699±321)vs(710±331)ng·mL-1、0到24 h的血浆浓度-时间曲线下面积AUC0-24为(2 687.3±1 188.8)vs(3 015.0±1 550.2)ng·h·mL-1、表观口服清除率CL/F为(51.3±23.8)vs(49.6±26.3)L·h-1。结论:建立的基于UHPLC-MS/MS测定人血浆中P-糖蛋白底物非索非那定浓度的分析方法灵敏、简单,并成功应用于灯盏花素对人P-糖蛋白体内活性影响的研究。OBJECTIVE To develop a simple,sensitive and specific UHPLC-MS/MS method for the determination of the Pglycoprotein substrate fexofenadine in human plasma,and to study the effect of breviscapine on the activity of human P-glycoprotein.METHODS Sample processing was accomplished through a simple protein precipitation with 500μL of acetonitrile to200μL plasma sample.The analytes were separated on a Waters ACQUITY UPLC？BEH C18 column（50 mm×2.1 mm,1.7μm）using a mobile phase consisting of acetonitrile（A）-0.1%formic acid in water（B）by gradient elution.The flow rate was0.3 mL·min-1.The ionization was performed using electrospray ionization in positive ion（ESI＋）by multiple reaction monitoring（MRM）mode for fexofenadine and diphenhydramine.The quantitative ion pairs of fexofenadine and diphenhydramine（internal standard）were m/z 502.2→466.5 and m/z 256.1→167.1,respectively.RESULTS The retention times of fexofenadine and diphenhydramine were 2.19 min and 1.89 min,respectively.The mean extraction recovery,mean process efficiency and mean matrix effect of fexofenadine were 84.3%-89.6%,86.3%-90.8% and 101.2%-104.2%,respectively.The calibration curve was linear in the range of 1.0-1 000 ng·mL-1（r=0.999 5）.Intra-day and inter-day RSD% of precision were≤10.9%,RE% of accuracy were in the range of-7.6%-5.0%,RSD% and RE% of stability were within 15%.After 18 volunteers were administrated with breviscapine or placebo,the peak concentration（Cmax）,area under the plasma concentration time curve from0 to 24 h（AUC0-24）and apparent oral clearance（CL/F）of fexofenadine were（699±321）vs.（710±331）ng·mL-1,（2 687.3±1 188.8）vs.（3 015.0±1 550.2）ng h·mL-1 and（51.3±23.8）vs.（49.6±26.3）L·h-1,respectively.CONCLUSION The method based on UHPLC-MS/MS for the determination of fexofenadine in human plasma is sensitive and simple,and is successfully applied to the study of the effect of breviscapine on the activity of human P-glycoprotein.

关 键 词：非索非那定 超高效液相色谱-串联质谱法 药动学 

分 类 号：R969[医药卫生—药理学]