UPLC-MS/MS法测定人血浆中普拉克索浓度及2种缓释片剂的生物等效性研究  

Determination of pramipexole in human plasma by UPLC-MS/MS and the bioequivalence study of two sustained release tables

作　　者：魏伯平 李莎 张敬尧 文琪 李鑫鑫 袁军 曾实 WEI Bo-ping;LI Sha;ZHANG Jing-yao;WEN Qi;LI Xin-xin;YUAN Jun;ZENG Shi(Bioanalytical Service Center of Sichuan Institute for Drug Control,NMPA Key Laboratory for Technical Research on Drug Products in Vitro and in Vivo Correlation,Chengdu 611731,China)

机构地区：[1]四川省药品检验研究院生物样本检测中心,药物制剂体内外相关性技术研究重点实验室,成都611731

出　　处：《药物分析杂志》2021年第4期619-626,共8页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:建立简便灵敏的液相色谱-串联质谱(LC-MS/MS)法测定人血浆中普拉克索浓度,并应用于2种缓释制剂生物等效性研究。方法:血浆样本加入内标,加入氢氧化钠后用乙酸乙酯进行液液萃取,取上清液氮气吹干,复溶离心后进样检测。采用Waters ACQUITY UPLC®HSS T3色谱柱(2.1 mm×50 mm,1.8μm),柱温40℃,以20 mmol·L^(-1)甲酸铵水溶液(A)-乙腈(B)为流动相,梯度洗脱,流速0.3 mL·min^(-1),以正离子MRM模式测定普拉克索(m/z 212.2→153.0)的浓度,普拉克索-d5二盐酸盐(m/z 217.0→153.0)作为内标,离子源为ESI源。结果:建立并验证了简便灵敏的UPLC-MS/MS法测定人血浆中普拉克索浓度,并应用于2种缓释制剂生物等效性研究。方法线性范围为5~1000 pg·mL^(-1),定量下限为5 pg·mL^(-1),质控样品批内、批间精密度(RSD)<8.5%,准确度相对偏差在标示值-7.67%~7.35%范围内,提取回收率、专属性、基质效应、稳定性等各项指标均符合国家药品监督管理局的技术指导原则。本法被成功应用于健康受试者单剂口服0.375 mg盐酸普拉克索缓释片的生物等效性研究,参比制剂空腹及餐后给药状态下平均Cmax分别为396.50和502.76 pg·mL^(-1),受试制剂空腹及餐后给药状态下平均Cmax分别为412.56和472.36 pg·mL^(-1)。结论:该方法具有前处理过程简单,灵敏度高,色谱峰形好的优点。盐酸普拉克索缓释片与其参比制剂相比,具有生物等效性。Objective:To establish a fast and sensitive LC-MS/MS method for determination of pramipexole in human plasma and to investigate the bioequivalence between 2 formulations of sustained release tablets.Methods:After addition of internal standard and sodium hydroxide,plasma was extracted by ethyl acetate.The supernatant was transferred and evaporated to dryness under a stream of nitrogen.The residue was re-dissolved,and centrifuged before sample injection.UPLC-MS/MS was performed on an Waters ACQUITY UPLC®HSS T3(2.1 mm×50 mm,1.8μm)column with the mobile phase consisting of 20 mmol·L^(-1)ammonium formate-water(A)and acetonitrile(B)in gradient elution.The flow rate was controlled at 0.3 mL·min^(-1)and the column temperature was set at 40℃.A mass spectrometer equipped with electrospray ionization source was used and pramipexole(m/z 212.2→153.0)was monitored in positive ion MRM mode,with pramipexole-d5 dihydrochloride(m/z 217.0→153.0)as internal standard.Results:A fast and sensitive UPLC-MS/MS method for determination of pramipexole in human plasma was established and validated and was applied to bioequivalence study of 2 kinds of sustained release tablets.The standard curves of pramipoxole were linear from 5 to 1000 pg·mL^(-1)and the lower limit of quantification was 5 pg·mL^(-1).The intra-day and inter-day relative standard deviation of qunlity-control samples was less than 8.5%,and the accuracy was in the range of-7.67%and 7.35%in terms of relative error.Recovery,specificity,matrix effect and stability met the guiding principles and criteria of NMPA.The method was successfully applied to a bioequivalence study of pramipexole dihydrochloride sustained release tablets containing 0.375 mg in healthy volunteers.The average Cmax under fasting and fed condition of the reference tablet were 396.50 and 502.76 pg·mL^(-1).The average Cmax under fasting and fed condition of the test tablet were 412.56 and 472.36 pg·mL^(-1),separately.Conclusion:The method is sensitive and simple in process,producing well chromatog

关 键 词：普拉克索 非麦角多巴胺激动剂 帕金森病 血浆浓度 生物等效性 液质联用 

分 类 号：R917[医药卫生—药物分析学]