LC-MS/MS测定人血浆中阿帕替尼的浓度及临床应用

Determination of apatinib in human plasma by LC-MS/MS and its clinical application

目的用LC-MS/MS建立快速检测人血浆中阿帕替尼浓度的方法,并应用于临床血药浓度监测。方法血浆样本用甲醇沉淀蛋白,以马来酸氯苯那敏为内标,色谱柱:ZORBAX SB-C18柱(100.0 mm×2.1 mm,3.5μm);流动相:甲醇(含0.1%甲酸)-0.1%的甲酸溶液,梯度洗脱,流速:0.4 mL·min^-1;用电喷雾离子源,正离子模式检测。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、稳定性。结果阿帕替尼的线性范围为25~2000 ng·mL^-1(r=0.9991),定量下限25 ng·mL^-1,日内RSD均<10.10%,日间RSD均<10.01%,绝对回收率>94.0%。50,500和1000 ng·mL^-1阿帕替尼的基质效应分别为(96.76±1.83)%,(95.00±3.22)%和(97.34±1.58)%,内标的基质效应为(97.71±4.12)%,样品在采集、储存、预处理及待检测过程中均能保持稳定。结论该方法操作简便,结果准确可靠,适用于人血浆中阿帕替尼的临床血药浓度监测。

Objective To establish a rapid method for the determination of apatinib in human plasma by LC-MS/MS and apply it to clinical plasma concentration monitoring.Methods Chlorpheniramine maleate was used as the internal standard.The analyte and internal standard were extracted from plasma by protein precipitation with methanol and separated on ZORBAX SB-C18 column(100.0 mm×2.1 mm,3.5μm).The mobile phase consisted of methanol(containing 0.1%formic acid)and 0.1%formic acid solution,gradient elution was used and the flow rate was 0.4 mL·min^-1.Electrospray ionization source was applied and operated in positive ion mode.The specificity,standard curve and quantitative lower limit,precision and recovery,matrix effect and stability of the method were investigated.Results The linear range of apatinib detection was 25-2000 ng·mL^-1(r=0.9991),the lower limit of quantification was 25 ng·mL^-1,the intra-day RSD was less than 10.10%,the inter-day RSD was less than 10.01%,and the absolute recovery was more than 94.0%.The matrix effects of the three concentrations of apatinib(50,500 and 1000 ng·mL^-1)were(96.76±1.83)%,(95.00±3.22)%and(97.34±1.58)%,and the matrix effect of internal standard was(97.71±4.12)%.The sample can be kept stable in the process of collection,storage,pretreatment and testing.Conclusion The method is simple,accurate and reliable,suitable for the clinical monitoring of apatinib in human plasma.