UPLC-MS/MS同时测定经典名方泽泻汤中6个成分的含量

Simultaneous Determination of Six Components in Classical Herbal Prescription Zexie Decoction by UPLC-MS/MS

目的:建立超高效液相色谱串联三重四级杆质谱法(UPLC-MS/MS)同时测定泽泻汤中泽泻醇A、泽泻醇B、23-乙酰泽泻醇B、白术内酯Ⅰ、白术内酯Ⅱ、白术内酯Ⅲ含量的方法。方法:泽泻汤按照经典名方自制,采用Agilent SB-C_(18)色谱柱(100 mm×2.1 mm,1.8μm),流动相为乙腈(A)-0.1%甲酸水溶液(B),梯度洗脱,体积流量为0.3 mL·min^(-1),柱温为40℃,进样量为1μL;质谱条件为电喷雾离子源(ESI),正离子模式采集,多反应监测(MRM)方式进行定量。结果:15 min内泽泻汤中6个成分的色谱峰分离良好,泽泻醇A、泽泻醇B、23-乙酰泽泻醇B、白术内酯Ⅰ、白术内酯Ⅱ、白术内酯Ⅲ分别在8.20~409.90(r=0.9993)、16.06~802.99(r=0.9995)、14.47~723.49(r=0.9999)、2.57~128.67(r=0.9996)、2.85~142.66(r=0.9985)、3.24~162.24 ng·mL^(-1)(r=0.9998)线性关系良好;平均回收率为95.39%~102.53%,RSD为0.71%~2.33%,精密度、稳定性、重复性符合要求。结论:所建立的方法简便、快速、准确,可为泽泻汤的质量标准提升提供参考。

Objective:To establish an ultra-high performance liquid chromatography/triple quadrupole mass spectrometry(UPLC-MS/MS)method for the simultaneous determination of alisol A,alisol B,23-O-acetylalisol B,atracylenolideⅠ,atracylenolideⅡ,and atracylenolideⅢin Zexie Decoction.Methods:Zexie Decoction was prepared according to the classical prescription.UPLC was performed on Agilent SB-C_(18)column(100 mm×2.1 mm,1.8μm)with the mobile phase consisting of acetonitrile(A)-water containing 0.1%formic acid(B)by gradient elution at a flow rate of 0.3 mL·min^(-1).The column temperature was set at 40℃and the volume of injection was 1μL.Multiple-reaction monitoring(MRM)scanning was used for quantification with the electrospray positive ion model.Results:The chromatographic peaks of six components in Zexie Decoction were well separated within 15 min.Alisol A,alisol B,23-O-acetylalisol B,atracylenolideⅠ,atracylenolideⅡ,and atracylenolideⅢshowed good linearity in linear ranges of 8.20-409.90(r=0.9993),16.06-802.99(r=0.9995),14.47-723.49(r=0.9999),2.57-128.67(r=0.9996),2.85-142.66(r=0.9985),3.24-162.24 ng·mL^(-1)(r=0.9998),respectively.The average recoveries were between 95.39%and 102.53%with the RSDs of0.71%-2.33%.The precision,stability,and repeatability also met the analysis requirements.Conclusion:The established method for simultaneous determination of six components in Zexie Decoction is easy,rapid,and accurate,which can provide experimental references for the improvement of the quality control of Zexie Decoction.