UHPLC-MS/MS测定人血浆中依匹哌唑浓度及其生物等效性研究  

Determination of Brexpiprazole Concentration in Human Plasma by UHPLC-MS/MS and Its Bioequivalence Study

作　　者：李莎 宋佩颖 李鑫鑫 吴强 魏伯平 袁军 曾实 LI Sha;SONG Peiying;LI Xinxin;WU Qiang;WEI Boping;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国家药监局重点实验室,成都611731

出　　处：《中国现代应用药学》2023年第4期494-499,共6页Chinese Journal of Modern Applied Pharmacy

摘　　要：目的建立简便灵敏的超高效液相色谱-串联质谱法(ultra performance liquid chromatography tandem mass spectrometry,UHPLC-MS/MS)测定人血浆中依匹哌唑浓度,并应用于2种片剂的生物等效性研究。方法采用Waters Acquity UPLC BEH C18色谱柱(2.1 mm×50 mm,1.7μm),以0.1%甲酸水溶液(A)-乙腈-甲醇(50∶50,含0.1%甲酸)(B)梯度洗脱,流速0.4 mL·min^(-1),进样量为2μL,柱温40℃,以正离子MRM模式测定依匹哌唑(m/z 434.2→273.2)的浓度,依匹哌唑-d8(m/z 442.4→281.3)作为内标,离子源为ESI源。血浆样本加入内标,加入甲醇后进行蛋白沉淀,取上清液稀释后进样检测。结果依匹哌唑在0.2~50 ng·mL^(-1)呈线性关系,定量下限为0.2 ng·mL^(-1),质控样品批内、批间精密度CV≤5.0%,准确度相对偏差在标示值–1.7%~5.5%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2 mg依匹哌唑口崩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19.62和20.83 ng·mL^(-1),受试制剂空腹及餐后给药状态下平均Cmax分别为21.68和19.74 ng·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。受试制剂依匹哌唑口崩片与其参比制剂相比,具有生物等效性。OBJECTIVE To establish a fast and sensitive UHPLC-MS/MS method for determination of brexpiprazole in human plasma and to investigate the bioequivalence between 2 formulations of tablets.METHODS Waters Acquity UPLC BEH C18(2.1 mm×50 mm,1.7μm)column was used with the mobile phase consisting of 0.1%formic acid water(A)and acetonitrile-methanol(50∶50,containing 0.1%formic acid)(B)in gradient elution.The flow rate was controlled at 0.4 mL·min^(-1)with the injection volume of 2μL and the column temperature was set at 40℃.A mass spectrometer equipped with electrospray ionization source was used and brexpiprazole(m/z 434.2→273.2)was monitored in positive ion MRM mode,with brexpiprazole-d8(m/z 442.4→281.3)as internal standard,ion source was ESI source.After addition of internal standard,plasma protein was precipitated by methanol,and supernatants were detected after dilution.RESULTS Brexpiprazole was linear in the range of0.2-50 ng·mL^(-1)and the lower limit of quantification was 0.2 ng·mL^(-1).The intra-day and inter-day precision CV of quality-control samples was≤5.0%,and the accuracy was in the range of–1.7%-5.5%in terms of relative error.Recovery rate,specificity,matrix effect and stability met the guiding principles and criteria of NMPA.The method was successfully applied to a bioequivalence study of brexpiprazole orally disintegrating tablets containing 2 mg in healthy volunteers.The average Cmaxunder fasting and fed condition of the reference tablet were 19.62 and 20.83 ng·mL^(-1).The average Cmaxunder fasting and fed condition of the test tablet were 21.68 and 19.74 ng·mL^(-1).CONCLUSION The method is sensitive and simple in process,and can produce well chromatographic performance.The test brexpiprazole orally disintegrating tablet is bioequivalent to the reference tablets.

关 键 词：依匹哌唑 抗精神病药 血浆浓度 生物等效性 超高效液相色谱-串联质谱法 

分 类 号：R917[医药卫生—药物分析学]