高效液相色谱-质谱联用法测定人血浆中安罗替尼浓度

Determination of anlotinib in human plasma by HPLC-MS/MS

目的建立测定人血浆中安罗替尼浓度的高效液相色谱-质谱联用方法,并用于安罗替尼胶囊在中国成年肿瘤患者的药代动力学研究。方法用蛋白沉淀法处理血浆,色谱柱:C18色谱柱(2.1 mm×50.0 mm,1.7μm),流动相:0.1%的甲酸水溶液-0.1%的甲酸甲醇溶液,梯度洗脱,流速:0.3 m L·min^-1,柱温:40℃,在多反应监测模式下,用电喷雾离子化源,正离子扫描模式检测。考察该方法的专属性、标准曲线与定量下限、准确度和精密度、基质效应和稳定性。结果安罗替尼血浆浓度在0.50~100.00 ng·m L^-1内线性关系良好,标准曲线为y=4.55×10^-2x+1.63×10^-3(r=0.9980),定量下限为0.50 ng·m L^-1,低、中、高3个质量浓度的质控样品日内、日间RSD均<10%,3种质量浓度质控样品的提取回收率在90.90%~103.50%,稳定性良好。结论该方法的准确度高、灵敏度高、重现性好,可用于人体中安罗替尼血浆浓度的测定。

Objective To establish a high performance liquid chromatography tandem mass spectrometry method(HPLC-MS/MS)for the determination of anlotinib in human plasma and use it for the pharmacokinetic study of anlotinib capsules in Chinese adult cancer patients.Methods The plasma was treated with protein precipitation,chromatographic column:C18 column(2.1 mm×50.0 mm,1.7μm),mobile phase:aqueous solution containing 0.1%formic acid-methanol solution containing 0.1%formic acid,gradient elution,column temperature:40℃,electrospray ionization source was applied and operated in multiple reaction monitoring mode via positive ionization.The specificity,standard curve and lower limit of quantitation,precision and recovery rate,matrix effect as well as the stability were investigated.Results The calibration curve was linear in the range of 0.50 to 100.00 ng·mL^-1,with the lower limit of quantitation of 0.50 ng·mL^-1.The standard curve of anlotinib was y=4.55×10^-2x+1.63×10^-3(r=0.9980).Intra-day and inter-day presions of low,medium and high quality control samples were all less than 10%,the average recoveries of them were between 90.90%-103.50%,and stability was good.Conclusion The established HPLC-MS/MS method was accurate,quick and sensitive,which can be used for the clinical pharmacokinetic studies of anlotinib in human plasma.