在线固相萃取LC-MS/MS法测定人全血中依维莫司浓度的不确定度评定

目的:评定在线固相萃取LC-MS/MS法测定人全血中依维莫司浓度的不确定度。方法:对全血依维莫司浓度测定过程中各影响因素,包括测定精密度、称量、对照品溶液的配制、标准含药全血样品的配制、在线固相萃取、仪器、标准曲线拟合等进行分析评定,用A类评定程序评价了分析过程中随机效应引起的不确定度,用B类评定程序评价了分析过程的其他因素引起的不确定度,最后根据各分量计算出合成不确定度并进行了扩展计算各变量的不确定度和合成不确定度,最终计算扩展不确定度。结果:人全血中低(0.520 ng·mL^-1),中(4.99 ng·mL^-1),高(84.99 ng·mL^-1)质量浓度依维莫司的扩展不确定度分别为0.056、0.40、7.16 ng·mL^-1(P=95%,k=2)。结论:在线固相萃取LC-MS/MS法测定人全血中依维莫司浓度的不确定度主要由提取回收率(尤其是低浓度)、对照品溶液配制、仪器允差及测定精密度引入。

Objective:To evaluate the uncertainty of everolimus in human blood by online solid-phase extraction LC-MS/MS.Method:The uncertainty caused by various factors in the whole process of determination, including repeatability, weighing, reference solution preparation, blood sample with reference substance preparation, online solid-phase extraction process, the apparatus and calibration fitting was estimated.The uncertainty caused by random effects was evaluated with type A and others were with type B.The combined uncertainty was calculated with all the components.Results:The expanded uncertainty for low(0.520 ng·mL^-1),medium(4.99 ng·mL^-1)and high(84.99 ng·mL^-1)level of everolimus was 0.056 ng·mL^-1,0.40 ng·mL^-1,and 7.16 ng·mL^-1, respectively(P=95%,k=2).Conclusion:The uncertainty of this method was mainly caused by extraction recovery(especially low level of sample),reference solution preparation,LC-MS/MS error and repeatability.