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山豆根中黄酮类成分的LC/MS定性分析与HPLC含量测定

Qualitative and quantitative analysis of flavonoids in Sophora tonkinensis by LC/MS and HPLC

  • 摘要: 目的:建立山豆根中黄酮类成分的定性、定量分析方法,完善药材质量控制手段。方法:以HPLC-DAD-ESI-MS方法,对山豆根中黄酮类成分进行在线分离与定性鉴定;以HPLC方法同时测定山豆根药材中5个主要活性黄酮的含量。结果:从山豆根中共鉴定了17个黄酮成分,其中7个成分的保留时间、紫外吸收特征和质谱碎片与对照品一致,分别是三叶豆紫檀苷、槲皮素、芒柄花素、高丽槐素、苦参酮、山豆根酮和山豆根色烯素;另外根据紫外吸收特征和质谱碎片并结合文献数据初步确定了10个成分的化学结构。建立了山豆根中三叶豆紫檀苷、槲皮素、高丽槐素、山豆根酮和山豆根色烯素等5个黄酮类成分的回归方程,线性关系均良好(r0.999 8),加样回收率为96.40%-104.43%,采用此方法成功检测了17个山豆根样品中上述5种黄酮成分的含量。结论:本文建立的方法可定性、定量分析山豆根中的黄酮类成分,可用于山豆根药材的质量分析。

     

    Abstract: AIM:To develop analytical methods for the identification and determination of the flavonoids in Sophora tonkinensis for comprehensive quality evaluation.METHODS:An HPLC-DAD-ESI-MS method was used for the separation and characterization of the flavonoids in S.tonkinensis,and a liquid chromatographic method was employed to simultaneously determine five major active flavonoids in this crude drug.RESULTS:Seventeen flavonoids were identified,among which,seven were unambiguously identified as trifolirhizin,quercetin,formononetin,macckiain,kurarinone,sophoranone,and sophoranochromene by comparing their retention times,and UV and MS spectra with those of the authentic compounds,and the other ten flavonoids were tentatively identified by comparing their UV and MS/MS spectra with those of literature data.Furthermore,five major active flavonoids,including trifolirhizin,quercetin,maackiain,sophoranone,and sophoranochromene were determined in S.tonkinensis.All calibration curves expressed good linearity(r0.999 8) within the test ranges,and the recovery from this method was 96.40%-104.43%.The developed HPLC method was successfully applied to quantitatively determine the five flavonoids in seventeen samples of S.tonkinensis.CONCLUSION:The developed method rapidly characterized the bioactive flavonoids of S.tonkinensis,and could be readily utilized to enhance the quality assurance approaches for this traditional Chinese medicine.

     

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