基于特征图谱和UPLC-Q-TOF/MS技术的北柴胡及其常见混淆品的化学鉴别研究

Study on Chemical Identification of Bupleurum Chinense and Counterfeits Based on Characteristic Chromatogram and UPLC-Q-TOF/MS Technology

  • 摘要:
    目的 基于特征图谱和UPLC-Q-TOF/MS技术对北柴胡及其常见混淆品进行化学鉴别。
    方法 对收集的不同基原、不同产地北柴胡及其常见混淆品进行薄层色谱鉴别,然后依据建立的HPLC特征图谱对北柴胡及其混淆品进行化学成分比较,进一步结合UPLC-Q-TOF/MS技术找出柴胡属混淆品代表性差异标志物。
    结果 薄层色谱显示,不同基原柴胡饮片均具有柴胡皂苷A和柴胡皂苷D,根据荧光斑点强度和位置可将其进行区分。北柴胡及其醋炙品特征图谱分别标定了14、16个共有峰,并指认其中6个成分。不同产地北柴胡饮片所含化学组分类似,建立的方法可以较好地体现柴胡特征。进一步通过特征图谱比较发现,竹叶柴胡在成分类群组成上与正品北柴胡接近,锥叶柴胡和大叶柴胡具有各自响应值较高的特征峰,藏柴胡皂苷含量明显高于北柴胡。利用UPLC-Q-TOF/MS共鉴定出69种柴胡化合物,主要为三萜皂苷类成分,其次是黄酮类,少数色酮、苯丙素类和炔类成分。聚类分析结果显示柴胡种间差异较为明显,不同柴胡混淆品具有代表性差异标志物。
    结论 建立的特征图谱和UPLC-Q-TOF/MS分析方法可用于北柴胡及其常见混淆品的化学鉴别研究,为全面合理开发利用柴胡资源提供依据。

     

    Abstract:
    OBJECTIVE To identify Bupleurum chinense pieces and its counterfeits based on the characteristic chromatogram and UPLC-Q-TOF/MS technology.
    METHODS Thin layer chromatography was used to identify Bupleurum chinense and its counterfeits collected from different origins and different producing areas. Then, the chemical constituents of Bupleurum chinense and its counterfeits were compared according to the established HPLC characteristic chromatogram, and the representative differential markers of Bupleurum chinense counterfeits were further identified by UPLC-Q-TOF/MS technology.
    RESULTS Thin layer chromatography showed that different original Bupleurum chinense pieces had saikosaponin A and saikosaponin D, which could be distinguished according to the intensity and position of fluorescent spots. There were 14 and 16 common peaks in the specific chromatogram of Bupleurum chinense and its vinegar-processed products respectively, and 6 components were identified. The chemical components of Bupleurum chinense pieces from different producing areas were similar, and the established method could better reflect the characteristics of Bupleurum chinense. Further comparison of the specific chromatogram of Bupleurum chinense and its counterfeits showed that the composition of B. marginatum Wall. was close to that of the authentic Bupleurum chinense., B. bicaule Helm and B. longiradiatum Turcz. had their own characteristic peaks with high response values. The content of saponins in B. marginatum var. stenophyllum was significantly higher. A total of 69 Bupleurum compounds were identified by UPLC-Q-TOF/MS mainly triterpenoid saponins, followed by flavonoids, a few chromones, phenylpropanoids and alkynes. The results of cluster analysis showed that the interspecific differences of Bupleurum chinense were obvious, and different Bupleurum counterfeits had representative differential markers.
    CONCLUSION The established characteristic chromatogram and UPLC-Q-TOF/MS method can be used for the chemical identification of Bupleurum chinense and its counterfeits, which provide a basis for the comprehensive and rational development and utilization of Bupleurum resources.

     

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