UHPLC-MS/MS法测定人尿液中2,4-二氨基-N^(10)-甲基蝶酸浓度的不确定度评定

Evaluation of uncertainty for deoxyaminopteroic acid analysis in human urine by UHPLC-MS/MS

目的对人尿液中2,4-二氨基-N^(10)-甲基蝶酸(DAMPA)浓度的超高效液相色谱质谱联用(UHPLC-MS/MS)定量分析方法的测量不确定度进行评定。方法通过对人尿液中DAMPA浓度测定方法分析,考察不确定度来源,应用已建立的该方法学数据计算不确定度。结果人尿液中DAMPA低浓度0.06μmol·L^(-1)及高浓度3μmol·L^(-1)质控样品的扩展不确定度分别为6.9×10^(-3)μmol·L^(-1)和0.21μmol·L^(-1)(P=95%,k=2)。结论UHPLC-MS/MS法测定人尿液中DAMPA浓度的不确定度主要由低浓度质控样品校正曲线拟合,基质效应和回收率引入。

Objective To set out the procedures for evaluation of uncertainty for 2,4-diamino-N^(10)-methylpteroic acid(DAMPA)analysis in human urine by UHPLC-MS/MS.Methods Detection process was analyzed.Uncertainty sources were established,then calculated each component of uncertainty,combined uncertainty and expaned uncertainty.Results The expanded uncertainty of methotrexate at low(0.06μmol·L^(-1))and high(3μmol·L^(-1))levels were 6.9×10^(-3) μmol·L^(-1) and 0.21μmol·L^(-1),respectively(P=95%,k=2).Conclusion The uncertainty of this UHPLC-MS/MS method was mainly caused by curve fitting,recovery and matrix effect at low level.