高效液相色谱质谱法测定人血浆中索拉菲尼的浓度

Determination of sorafenib in human plasma by liquid chromatography tandem mass spectrometry

目的建立人血浆中索拉菲尼浓度测定的HPLC-MS/MS方法。方法人血浆样品用乙腈-沉淀蛋白后,色谱柱:Eclipse Plus C18(2.1 mm×100 mm,3.5μm),流动相:乙腈∶10 mmol·L^(-1)醋酸铵溶液(均含0.05%甲酸)=80∶20,流速:0.25 m L·min^(-1)。用电喷雾离子化源,正离子方式,多反应监测扫描方式进行监测。考察该方法的专属性、标准曲线和定量下限、精密度与回收率、基质效应以及稳定性。结果血浆中索拉菲尼的线性范围为0.02~10.00μg·m L^(-1)(r=0.997 1),定量下限为0.02μg·m L^(-1);质控样品的准确度在95.01%~104.86%;日内、日间RSD均小于15%;提取回收率为82.08%~89.72%,无明显基质效应。质控样品室温放置8 h、处理后自动进样器4℃放置12 h,反复冻融3次,于-80℃环境中放置20 d后准确度在91.78%~106.19%,RSD在4.31%~9.73%。结论本方法快速、灵敏、准确,专属性强,重复性好,适用于人血浆中索拉菲尼浓度的测定。

Objective To develop a HPLC-MS/MS method for determination of sorafenib in human plasma.Methods The plasma samples were precipitated by acetonitrile,then the supernatant were separated on Eclipse Plus C18column（2.1 mm × 100 mm,3.5 μm） and eluted with acetonitrile-10 mmol · L-1ammonium acetate（80 ∶ 20,containing0.05% formic acid）.Detection of the analyte was achieved using positive ion electrospray ionization（ESI） in the multiple reaction monitoring（MRM） mode.The validation parameters included selectivity,linearity,accuracy and precision,recovery,matrix effect and stability.Results The linear calibration curve of sorafenib was obtained in the concentration range of 0.02-10.00 μg·m L^（-1）（r = 0.997 1）,with the lower limit of quantitation of 0.02 μg·m L^（-1）.Accuracy was 95.01%-104.86%,intra-day and inter-day relative standard deviations were both below15%.The absolute recovery in plasma was 82.08%-89.72%,and no significant matrix effect was found.The QC plasma samples were stable in autosampler（24 h） at 4 ℃,on bench-top（8 h） at room temperature,through repeated three freeze/thaw cycles and under the frozen condition at-80 ℃ for 20 days.Conclusion The established method is rapid,sensitive,accurate,specific and reliable,and suitable for the determination of sorafenib in human plasma and pharmacokinetic study.