用LC-MS/MS法测定人血浆中乌帕替尼的浓度  

Determination of upatinib honcentration in human plasma by LC-MS/MS method

作　　者：高瑶瑶 田晓彤 王茉莉 闫凯 王柳 李姿 高燕霞 GAO Yao-yao;TIAN Xiao-tong;WANG Mo-li;YAN Kai;WANG Liu;LI Zi;GAO Yan-xia(College of Chemistry and Materials Science,Hebei University,Baoding 071002,Hebei Province,China;NMPA Key Laboratory for Quality Control and Evaluation of Generic Drug,Hebei Institute of Pharmaceutical and Medical Device Inspection,Shijiazhuang 050000,Hebei Province,China)

机构地区：[1]河北大学化学与材料科学学院,河北保定071002 [2]河北省药品医疗器械检验研究院,国家药品监督管理局仿制药质量控制与评价重点实验室,河北石家庄050000

出　　处：《中国临床药理学杂志》2025年第3期393-396,共4页The Chinese Journal of Clinical Pharmacology

基　　金：中央引导地方科技发展资金资助项目(226Z2402G)。

摘　　要：目的建立一种液相色谱-质谱联用(LC-MS/MS)法定量测定人血浆中乌帕替尼血药浓度的方法。方法以乌帕替尼-15N-d2为内标,用乙腈沉淀蛋白预处理。以Waters BEH C_(18)(2.4 mm×50.0 mm,1.7μm)色谱柱进行分离;以乙腈(含0.1%甲酸)和0.1%甲酸水溶液为流动相,梯度洗脱,流速为0.3 mL·min^(-1),柱温40℃。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/z 388.90→255.90(乌帕替尼),m/z 384.10→256.20(乌帕替尼-15N-d2,内标)。考察该方法的专属性、标准曲线、定量下限、精密度、回收率、稀释可靠性、基质效应和稳定性。结果血浆中的内源性成分、内标与乌帕替尼之间均无相互干扰。质量浓度0.75~60.00 ng·mL^(-1)乌帕替尼与峰面积的线性关系良好(r=0.9986),标准曲线方程为y=8.24×10^(-2)x+0.61×10^(-2),定量下限为0.75 ng·mL^(-1),质控样品批内、批间精密度变异系数均≤5.3%;5倍稀释后的样品浓度准确度为1.1%,变异系数为4.6%。低、高质量浓度血浆质控样品的基质效应因子分别为1.1和1.0。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乌帕替尼的浓度,可用于其在健康人体中的药代动力学及生物等效性研究。Objective To establish liquid chromatography-mass spectrometry(LC-MS/MS)quantitative method for determining the blood concentration of Upatinib in human plasma.Methods Upatinib15N-d2 was used as the internal standard,and acetonitrile was used for protein precipitation pretreatment.Separate using a Waters BEH C_(18)(2.4 mm×50.0 mm,1.7μm)chromatographic column;acetonitrile(containing 0.1%formic acid)and 0.1%formic acid aqueous solution were used as mobile phases,with gradient elution at a flow rate of0.3 mL·min^(-1)and column temperature of 40℃.The ion pair used for quantitative analysis was m/z 388.90→255.90(Upatinib)and m/z384.10→256.20(Upatinib-15N-d2,internal standard)respectively by using an electrospray ion source and positive ion multi reaction monitoring mode scanning.The specificity,standard curve,quantitative cutoff,precision,recovery rate,dilution reliability,matrix effect and stability of the method were examined.Results There is no mutual interference between endogenous components,internal standards and Upatinib in plasma.The linear relationship between the mass concentration of 0.75-60.00 ng·mL^(-1)Upatinib and peak area is good(r=0.9986),the standard curve equation was y=8.24×10^(-2)x+0.61×10^(-2),with a lower limit of quantification of0.75 ng·mL^(-1).The intra-day and inter-day batch precision coefficients of variation of the quality control samples are both≤5.3%.The accuracy of the sample concentration after 5-fold dilution is 1.1%,and the coefficient of variation is 4.6%.The matrix effect factors of low and high-quality plasma quality control samples are 1.1 and 1.0,respectively.The stability of whole blood samples and plasma samples meets the requirements of biological sample analysis.Conclusion The method used is simple,rapid,highly accurate,and sensitive,and is suitable for determining the concentration of Upatinib in human plasma.It can be used for pharmacokinetic and bioequivalence studies of Upatinib in healthy individuals.

关 键 词：乌帕替尼 液相色谱-质谱联用 血药浓度 

分 类 号：R971.1[医药卫生—药品]