UPLC-MS/MS法测定人血浆中的阿哌沙班  

Determination of Apixaban in human plasma by UPLC-MS/MS

作　　者：唐维英 杨姗姗 魏伯平 吴强 袁军 朱恒怡 TANG Weiying;YANG Shanshan;WEI Boping;WU Qiang;YUAN Jun;ZHU Hengyi(NMPA Key Laboratory for Technical Research on Drug Products in vitro and in vivo Correlation,Sichuan Institute for Drug Control,Chengdu,Sichuan,611731 P.R.China)

机构地区：[1]四川省药品检验研究院,国家药监局药物制剂体内外相关性技术研究重点实验室,四川成都611731

出　　处：《华西药学杂志》2024年第4期411-415,共5页West China Journal of Pharmaceutical Sciences

摘　　要：目的采用超高效液相色谱-串联质谱法测定人血浆中的阿哌沙班。方法采用乙腈沉淀蛋白预处理血浆样品,WatersXbridgeC_(18)色谱柱(50mm×2.1mm,3.5μm)进行分离,以5%乙腈(A)和95%乙腈(B)的含0.1%甲酸水溶液作为流动相,梯度洗脱,流速0.4mL·min^(-1),柱温40℃,进样量2μL。采用电喷雾离子源,正离子多反应监测模式扫描,用于定量分析的离子对分别为m/z460.3→443.3(阿哌沙班)、m/z464.4→447.3(^(13)C阿哌沙班-d_(3)内标)。结果0.5~400ng·mL^(-1)阿哌沙班与峰面积的线性关系良好(r=0.9995),定量下限为0.5ng·mL^(-1),提取回收率≥99.22%,质控样品的批内、批间RSD均≤4.53%;全血样品和血浆样品的稳定性符合生物样品分析的要求。结论所用方法简便快速、准确度高、灵敏度好,适用于测定人血浆中阿哌沙班的浓度,可用于其在健康人体中的药动学及生物等效性研究。OBJECTIVE To determine Apixaban in human plasma by UPLC-MS/MS.METHODS The samples were treated by protein precipitation with acetonitrile.Apixaban was separated on a Waters Xbridge C_(18)(50 mm×2.1 mm,3.5μm)chromatographic column using gradient elution with the mobile phase consisting of 5%acetonitrile(A)and 95%acetonitrile(B)solution(containing 0.1%formic acid).The flow rate was set at 0.4 mL·min^(-1),the column temperature was set at 40℃,and the injection volume was 2μL.The detection of the analytes was performed on electrospray ionization in positive mode by multiple monitoring with the transition of m/z 460.3→443.3(Apixaban)and m/z 464.4→447.3(^(13)C Apixaban-d_(3),IS).RESULTS A good linearity was observed for the Apixaban in the range of 0.5-400 ng·mL^(-1)(r=0.9995),the lower limit of quantification was 0.5 ng·mL^(-1),the recovery of Apixaban was≥99.22%,and the intra-day and inter-day RSD of quality-control samples were≤4.53%.The stability of whole blood and plasma samples met the requirements of biological sample analysis.CONCLUSION The method is easy,rapid,accurate and sensitive.It is suitable for the determination of Apixaban concentration in human plasma,and can be used to study the pharmacokinetics and bioequivalence of Apixaban tablets in healthy human subjects.

关 键 词：阿哌沙班 抗凝血药 超高效液相色谱-串联质谱法 血浆浓度 蛋白沉淀法 药动学 生物等效性 方法验证 

分 类 号：R917[医药卫生—药物分析学]