LC-MS/MS法测定人血浆中苦参碱浓度的不确定度评定

Uncertainty Evaluation for Matrine Determination in Human Plasma by LC- MS / MS

目的:评定高效液相色谱-串联质谱法(LC-MS/MS)法测定人血浆中苦参碱浓度的不确定度。方法:对苦参碱浓度测定过程中各影响因素,包括测定精密度、称量、溶液的配制、含药血浆的配制、血浆蛋白沉淀、仪器、标准曲线拟合等进行分析评定,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:人血浆中3.0 ng·m L-1(2.938 ng·m L-1)和30.0 ng·m L-1浓度苦参碱的扩展不确定度分别为(2.938±1.091)ng·m L-1、(31.18±2.712)ng·m L-1(P=95%,k=2)。结论:LC-MS/MS法测定人血浆中苦参碱浓度的不确定度主要由标准曲线(尤其是低浓度)、提取回收率和血浆样品制备处理及仪器允差引入。

Objective： To evaluate the uncertainty of matrine determination in human plasma by LC- MS / MS. Methods： The uncertainty caused by various factors in the whole process of determination,including repeatability,weighing,solution preparation,protein precipitation process,the apparatus and calibration fitting,was estimated. The expanded uncertainty was obtained by calculating each component of uncertainty and combined uncertainty. Results： The expanded uncertainty for 3. 0 ng·m L^- 1 and 30. 0 ng·m L^-1 of matrine was（ 2. 938 ± 1. 091） ng·m L^- 1,（ 31. 18 ± 2. 712） ng·m L^- 1（ P = 95%,k = 2）. Conclusion： The uncertainty of this method was mainly caused by calibration curve（ especially low level of sample）,extraction recovery and plasma preparation,sample pretreatment and LC- MS / MS error.