高效液相色谱串联质谱法测定非小细胞肺癌患者血浆中阿法替尼浓度

Determination of afatinib in plasma of non-small cell lung cancer patients by liquid chromatography tandem mass spectrometry

目的建立测定人血浆中阿法替尼浓度的LC-MS/MS方法,并用于非小细胞肺癌患者血药浓度测定。方法以阿法替尼-d6为内标,血浆样品经乙腈沉淀蛋白,色谱柱:Eclipse XDB-C18(2.1 mm×100.0 mm,3.5μm),流动相:乙腈含0.1%氨水-10 mmol·L^(-1)醋酸铵含0.1%氨水,梯度洗脱,流速:0.30mL·min^(-1),柱温:25℃,进样量:20μL。用电喷雾离子化源(ESI),正离子方式,多反应监测模式。考察该方法的专属性、标准曲线与定量下限、精密度与准确度、基质效应。结果阿法替尼在0.50~100.00μg·L^(-1)内,线性关系良好(r=0.996 7),定量下限为0.50μg·L^(-1);批内和批间精密度良好,RSD均小于15%,相对误差在-5.52%~6.28%内;阿法替尼和内标的提取回收率> 80%,且不存在明显的基质效应。结论本方法特异性强,灵敏度高,可用于人血浆中阿法替尼浓度的分析。

Objective To devolope a liquid chromatography-tandem mass spectrometry(LC-MS/MS) method for the determination of afatinib in human plasma and apply it to the patients with non-small cell lung cancer.Methods Afatinib-d6 was employed as the internal standard.The plasma samples were precipitated by acetonitrile and chromatographed on Agilent Eclipse XDB-C18 column(2.1 mm×100.0 mm,3.5 μm).Mobile phase consisted of acetonitrile-10 mmol·L^(-1)ammonium acetate(both of containing 0.1% ammonia) at the flow rate of0.3 mL·min^(-1),and the column temperature was 25 ℃.The whole analytical time was 3 min.Electrospray ionization(ESI) source was applied and operated in the positive multiple reaction monitoring mode.The selectivity standard curve,precision and accuracy,extraction recoveries,matrix effect and stability were investigated.Results The standard curve was demonstrated to be liner in the range of 0.50-100.00 μg·L^(-1)(r=0.996 7),with the lower limit of quantitation at0.50 μg·L^(-1).The RSDs of inter-day and intra-day were less than15%,and accuracy were within-5.52%-6.28%.The average recoveries were between 81.75% and 90.52%.There was no obvious matrix effect.Conclusion The method is specific and sensitive,suitable for the determination of afatinib in NSCLC patients plasma efficiently.