HPLC-MS/MS法测定人血浆伪人参皂苷GQ浓度的不确定度评定

摘要

Objective To evaluate the uncertainty of the pseudo-ginsenoside GQ (PGQ) concentration in human plasma by HPLC-MS/MS.Methods The whole process of PGQ determination by HPLC-MS/MS in human plasma was evaluated and the uncertainty caused by repeatability,weighing,standard solution preparation,biological sample preparation,extraction recovery process,recovery,instrument precision and calibration curve fitting were evaluated,respectively.The combined and expanded uncertainty values were both calculated.Results The expanded uncertainty values for low (15.16 ng·mL-1),medium (2 516.67 ng·mL-1) and high (3 902.00 ng·mL-1) levels of PGQ were 1.39,177.74 and 262.69 ng·mL-1,respectively (P =95 ％,k =2).Conclusion The uncertainty of the PGQ determination in human plasma by HPLC-MS/MS is mainly caused by recovery,repeatabihty and sample preparation at low concentration,by sample preparation and recovery at medium and high concentration.%目的 评估高效液相色谱-质谱联用法(HPLC-MS/MS)测定人血浆伪人参皂苷GQ (PGQ)浓度的不确定度.方法 全面分析HPLC-MS/MS法测定人血浆中PGQ浓度的整个过程,对测量重复性、标准品称量、工作液配制、生物样品配制、萃取回收过程、仪器允差和标准曲线拟和等引起的不确定度分别进行评定,计算合成不确定度并进行扩展.结果 人血浆低浓度(15.16 ng·mL-1)、中浓度(2 516.67 ng·mL-1)和高浓度(3 902.00 ng· mL-1)PGQ的扩展不确定度分别为1.39,177.74和262.69 ng· mL-1(P=95％,k=2).结论 HPLC-MS/MS法测定人血浆中PGQ浓度的不确定度在低浓度时主要由萃取回收率、生物样品配制和测量重复性引入,在中浓度和高浓度时主要由生物样品配制和萃取回收率引入.