芪红水煎剂化学成分的HPLC-FT-ICR-MS快速表征与HPLC多成分的含量测定

Rapid Characterization of Chemical Constituents of Qihong Decoction by HPLC-FT-ICR-MS and Determination of Multicomponent by HPLC

目的采用高效液相色谱串联高分辨傅立叶变换离子回旋共振质谱(High performance liquid chromatography tandem high resolution Fourier transform ion cyclotron resonance mass spectrometry,HPLC-FTICR-MS)技术对芪红水煎剂的化学成分进行快速表征,建立高效液相色谱法(high-performance liquid chromatography,HPLC)对芪红水煎剂中多成分进行含量测定,分析黄芪、水红花子配伍对主要成分溶出的影响,完善其质量控制。方法HPLC-FT-ICR-MS采用电喷雾离子化源(Electrospray ionization source,ESI)负离子模式检测,根据高分辨数据及二级质谱信息结合对照品数据及相关文献对芪红水煎剂化学成分进行快速鉴定;芪红水煎剂中6种主要成分的HPLC含量测定采用Venusil MP-C18色谱柱(4.6 mm×250 mm,5μm),甲醇-0.5%磷酸溶液梯度洗脱,流速1.0 mL·min^-1,检测波长254 nm和290 nm。结果对芪红水煎剂提取物中的26个化学成分进行表征,其中9个成分来源于水红花子,17个成分来源于黄芪。与单味药相比,芪红水煎剂中毛蕊异黄酮-7-O-β-D-葡萄糖苷、N-反式-阿魏酰酪胺、槲皮素的煎出量均明显增加(P<0.01),花旗松素、芒柄花素的煎出量明显降低(P<0.01),毛蕊异黄酮煎出量变化不明显(P>0.05)。结论该方法可快速对芪红水煎剂中的主要成分进行定性分析,所建多成分含量测定方法可以反映水红花子和黄芪共煎后对主要成分溶出率的影响,为进一步研究其配伍规律提供理论依据。

Objective To establish a rapid method for characterizing the chemical constituents of Qihong Decoction by HPLC-FT-ICR-MS,and determination of multi-components in Qihong Decoction by HPLC,to clarify the effect of compatibility of Astragalus membranaceus and Fructus Polygoni orientalis on the dissolution rate of main components,perfecting its quality control method.Methods HPLC-FT-ICR-MS analysis was performed on electrospray ionization source(ESI)with negative ion mode detection.The components were identified based on high resolution data and the secondary mass spectrometry information,combined with reference material data and related literature;The contents of six compounds were determined using Venusil MP-C18 column(4.6 mm×250 mm,5μm)with mobile phase composed of methanol and 0.2%formic acid solution for gradient elution at the flow rate flow rate 1 mL·min^-1 by HPLC,the detection wavelengths were 254 nm and 290 nm.Results 26 constituents were identified in the extract of Qihong decoction.Among them,9 were from Fructus Polygoni orientalis and 17 were from Astragalus membranaceus.Compared with the single herb,the extraction yeild of calycosin glycoside,N-trans-Feruloyltyramine and quercetin in Qihong decoction increased significantly(P<0.01),while the contents of taxifolin and formononetin decreased significantly(P<0.01),and the change of calycosin content was not obvious(P>0.05).Conclusion This method could provide rapid qualitative analysis for the chemical components of Qihong decoction,the established HPLC method of determining the contents of multi-components could reflect the influence of the dissolution rate of the main components by mixed decoction of Astragalus membranaceus and Fructus Polygoni orientalis,which provides a basis for further study on the compatibility law.