UPLC-MS/MS法同时测定三七总皂苷中5种皂苷类成分在大鼠血浆中的含量

Simultaneous determination of 5 kinds of saponins in Panax notoginseng saponins in rat plasma by UPLC-MS/MS

目的建立超高效液相色谱-串联质谱(UPLC-MS/MS)法同时测定大鼠口服三七总皂苷后血浆中三七皂苷R1、人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1、人参皂苷Rd的含量。方法以柴胡皂苷A为内标,蛋白沉淀法对血浆样品进行处理。色谱柱:ACQUITY BEH C18柱(100 mm×2.1 mm,1.7μm),流动相:乙腈-0.1%甲酸水溶液,梯度洗脱,流速:0.4 mL·min^(-1)。采用电喷雾离子源,负离子模式及多反应监测模式。考察该方法的专属性、标准曲线与定量下限、精密度、提取回收率、基质效应和稳定性。结果UPLC-MS/MS法检测三七皂苷R1、人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1、人参皂苷Rd的标准曲线线性范围分别是2.01~1005.50,1.94~964.80,1.94~969.50,1.85~926.60,1.93~965.20 ng·mL^(-1),批内与批间精密度分别为0.96%~7.59%、0.77%~7.16%,提取回收率为85.01%~101.99%;基质效应为92.48%~105.81%;在设定的各种条件下稳定性良好。结论建立的UPLC-MS/MS分析方法简便、快捷,适用于大鼠血浆中三七皂苷R1、人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1、人参皂苷Rd的含量测定。

Objective To establish an ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method for the simultaneous determination of notoginsenoside R1,ginsenoside Rg1,ginsenoside Re,ginsenoside Rb1 and ginsenoside Rd in rat plasma after oral administration of Panax notoginseng saponins.Methods The plasma samples were treated by protein precipitation with saikosaponin A as the internal standard.The detection was performed on ACQUITY BEH C18(100 mm×2.1 mm,1.7μm)by the gradient elution with acetonitrile and 0.1%formic acid aqueous solution as mobile phase,at 0.4 mL·min^(-1)flow rate.The mass spectrometer was operated under the negitive ion mode with the electrospray ion source and the detection was performed by multiple reaction monitoring.The specificity,standard curves and limit of detection,precision,extraction recovery,matrix effect and stability of the method were investigated.Results The standard curves of notoginsenoside R1,ginsenoside Rg1,ginsenoside Re,ginsenoside Rb1 and ginsenoside Rd by UPLC-MS/MS were 2.01-1005.50,1.94-964.80,1.94-969.50,1.85-926.60,1.93-965.20 ng·mL^(-1),respectively.Intra-day and inter-day precision were 0.96%-7.59%and 0.77%-7.16%respectively,and the extraction recovery were 85.01%-101.99%.The matrix effect were 92.48%-105.81%.It has good stability under various conditions.Conclusion The established UPLC-MS/MS method is simple and fast,and suitable for the determination of notoginsenoside R1,ginsenoside Rg1,ginsenoside Re,ginsenoside Rb1 and ginsenoside Rd in rat plasma.