液相色谱-质谱串联法测定人血浆中羟尼酮浓度的不确定度评定

Uncertainty Evaluation for the Determination of Hydronidone in Human Plasma by LC-MS/MS

目的评定液相色谱-质谱串联(LC-MS/MS)法测定人血浆中羟尼酮浓度的不确定度。方法分析LC-MS/MS测定血浆羟尼酮过程中不确定度的来源,包括对照品的称量、标准溶液的配制、含药血浆样品的配制、血浆样品的处理、标准曲线的拟合、仪器误差、基质效应、重复性等,评定各来源分量的不确定度,计算合成不确定度和扩展不确定度。结果人血浆中低(6.0 ng·mL^-1)、中(60.0 ng·mL^-1)、高(300.0 ng·mL^-1)浓度羟尼酮的扩展不确定度分别为0.63,5.46,33.40 ng·mL^-1(k=2,P=95%)。结论 LC-MS/MS法测定人血浆中羟尼酮浓度的不确定度主要由基质效应、仪器误差、血浆样品的处理(提取回收率)、重复性(精密度)及标准曲线拟合(低浓度)引入。

Objective To evaluate the uncertainty for the determination of hydronidone concentration in human plasma by LC-MS/MS. Methods The uncertainty caused by various factors in the whole process of determination of hydronidone in human plasma by LC-MS/MS, including weighting, preparation of standard solutions, drug-spiked sample preparation, protein precipitation, calibration curve fitting, equipment error, matrix effect and repeatability was analyzed.The uncertainty of each source component, as well as the combined uncertainty and combined uncertainty were calculated. Results The expanded uncertainty for low(6.0 ng·mL^-1), medium(60.0 ng·mL^-1) and high(300.0 ng·mL^-1) level of hydronidone was 0.63, 5.46, 33.40 ng·mL^-1, respectively(k=2, P=95%). Conclusion The uncertainty for the determination of hydronidone in human plasma by LC-MS/MS is mainly caused by matrix effect, equipment error, pretreatment of plasma samples, repeatability, and calibration curve fitting(especially for the low concentration).