辛夷指纹图谱的研究
摘要
辛夷是我国的传统中药,为木兰科植物望春花Magnolia biondii Pamp.、玉兰Magnolia denuda ta Desv.或武当玉兰Magnolia sprengeri Pamp.的干燥花蕾。冬末春初花未开放时采收,除去枝梗,阴干。辛夷同时也是我省的道地药材,除挥发油外,还含有木脂素,生物碱等各类有效成分,具有消炎,降压等多种药理作用。
目前虽然已有其他学者制定了辛夷药材挥发油的气相色谱(GC)和气相色谱-质谱(GC-MS)指纹图谱,但在提取方法上多局限于传统的水蒸气蒸馏法,耗时长,且对药材的消耗量较大。本文采用的顶空-固相微萃取法(HS-SPME)操作简便快捷,取样量少,且无溶剂消耗,可为建立辛夷的GC指纹图谱提供一种更好的手段。而研究不同技术所得的指纹图谱之间的关系,建立辛夷的高效液相色谱(HPLC)指纹图谱,可让指纹图谱真正的反映辛夷的内在质量。文献中也尚未见建立辛夷的顶空-固相微萃取-气相色谱(HS-SPME-GC)指纹图谱和高效液相色谱指纹图谱研究的报道。因此研究辛夷药材的顶空固相微萃取-气相色谱指纹图谱和高效液相色谱指纹图谱,从而为辛夷药材的鉴别和质量控制提供方法和依据,具有十分重要的意义。
本文首次建立了辛夷的顶空-固相微萃取-气相色谱指纹图谱。首先采用顶空-固相微萃取技术直接提取辛夷中的挥发性成分,应用气相色谱进行分离,总共获得47个共有特征峰,经顶空-固相微萃取-气相色谱-质谱(HS-SPME-GC-MS)联用技术鉴定,确认出其中31个组分。然后采用SPSS统计软件对不同产地或不同采集时间的14批辛夷样品的指纹图谱数据进行聚类分析后,Ⅰ类中10批样品被选用来建立共有模式。以共有模式为参照,应用国家药典委员会《中药色谱指纹图谱相似度评价系统B版》分别计算各样品与共有模式之间的相似度。有两个批次的药材的相似度低于0.9,此结果与聚类分析结果一致,两种方法得到了相互验证,说明HS-SPME-GC指纹图谱可用于辛夷药材的质量评价。
同时,本文还首次建立了辛夷的HPLC指纹图谱。首先,在流动相的组成上,比较了不同流动相组成的分离效率,最终确定以水和乙腈作为流动相。其次在提取方法上,分别使用不同的提取溶剂,以提取效率为目标筛选,确定乙酸乙酯为最终提取溶剂。另外,在检测器方面,比较了蒸发光散射检测器和紫外检测器的差异。与蒸发光散射检测器相比,紫外检测器灵敏度高,出峰多。在优化的色谱条件下,各批辛夷药材的相似度均达到了0.9以上,符合指纹图谱的要求,为其资源品质的评价提供了更充分的科学依据。
The Magnolia Flos is our country's traditional traditional Chinese medicine, Plants the people's hopes for the wooden orchid aceae spring to spend Magnolia biondii Pamp., magnolia Magnolia denudata Desv. Or Magnolia sprengeri Pamp. dry flower bud. The winter end spring first flowering has not opened when recovers, except a stem, dries in the shade.
The Magnolia Flos simultaneously is also our province road place raw material for medicine, besides volatile oil, but also includes the wooden fat element, the alkaloid and so on each kind of effective component, has the sterilization, the voltage dropping and so on many kinds of pharmacological actions, although at present had other scholars to formulate Magnolia Flos raw material for medicine volatile oil GC and the GC-MS fingerprint, but limits in the extraction method to the traditional steam distillation, time-consuming is long, and is big to raw material for medicine and the resolver consumption, But regarding the Magnolia Flos in the other type's effective component, through establishes its highly effective liquid phase fingerprint atlas, between the research different technology obtained fingerprint atlas's relations, may let the fingerprint true reflection Magnolia Flos the intrinsic quality. In addition, in the literature also not yet sees the Magnolia Flos to go against the spatial solid phase micro extract- gas chromatography fingerprint and the highly effective liquid chromatography fingerprint report. Therefore the research raw material for medicine Magnolia Flos goes against the spatial solid phase micro extract-gas chromatography fingerprint atlas and the highly effective liquid chromatography fingerprint, provides the method and the basis for the Magnolia Flos raw material for medicine's distinction and the quality control, has the very vital significance.
This article established the Magnolia Flos to go against the spatial-solid phase micro extract-gas chromatography for the first time (HS-SPME-GC) the fingerprint, has provided one better method for system's science appraisal and the appraisal Magnolia Flos's quality. Using the HS-SPME technology directly extracted the volatile organic compounds in Magnoliae Flos, then these compounds be separated and identificated by GC and GC-MS. The Chromatographic fingerprint of Magnoliae Flos was established by HS-SPME-GC. The 47 common peaks were obtained and the 31 common peaks were displayed in the them by HS-SPME-GC-MS. The 14 samples were classified as 3 grades by cluster analysis and the 10 superior were confirmed to establish the mutual models, as a reference to it, the similarity were calculated between the samples and the mutual models, the results of cluster and similarity analysis remained the same. This research method for evaluating the quality of Magnoliae Flos provides scientific basis.
At the same time, this article also for the first time used the highly effective liquid phase chromatography (HPLC) to establish the Magnolia Flos Central Africa volatile constituent HPLC fingerprint, has provided the basis for the quality synthetic evaluation Magnolia Flos raw material for medicine's intrinsic quality. First, in the flowing composition, compared with the separation efficiency which the different flowing composed, determined finally took the flowing by the water and the methyl cyanide. Next in the extraction method, uses the different extraction resolver separately, take the extraction efficiency as goal screening, the definite ethyl acetate to withdraw the resolver finally. Moreover, in detector aspect, compared with evaporation light scattering detector and ultraviolet detector's difference. Compares with the evaporation light scattering detector, because the ultraviolet detector the sensitivity is higher than the peak to be many. Under the optimized chromatograph condition, each batch of Magnolia Flos raw material for medicine's similarity has achieved above 0.9, meets the fingerprint atlas requirement, has provided a fuller scientific basis for its resources quality's appraisal.
目前虽然已有其他学者制定了辛夷药材挥发油的气相色谱(GC)和气相色谱-质谱(GC-MS)指纹图谱,但在提取方法上多局限于传统的水蒸气蒸馏法,耗时长,且对药材的消耗量较大。本文采用的顶空-固相微萃取法(HS-SPME)操作简便快捷,取样量少,且无溶剂消耗,可为建立辛夷的GC指纹图谱提供一种更好的手段。而研究不同技术所得的指纹图谱之间的关系,建立辛夷的高效液相色谱(HPLC)指纹图谱,可让指纹图谱真正的反映辛夷的内在质量。文献中也尚未见建立辛夷的顶空-固相微萃取-气相色谱(HS-SPME-GC)指纹图谱和高效液相色谱指纹图谱研究的报道。因此研究辛夷药材的顶空固相微萃取-气相色谱指纹图谱和高效液相色谱指纹图谱,从而为辛夷药材的鉴别和质量控制提供方法和依据,具有十分重要的意义。
本文首次建立了辛夷的顶空-固相微萃取-气相色谱指纹图谱。首先采用顶空-固相微萃取技术直接提取辛夷中的挥发性成分,应用气相色谱进行分离,总共获得47个共有特征峰,经顶空-固相微萃取-气相色谱-质谱(HS-SPME-GC-MS)联用技术鉴定,确认出其中31个组分。然后采用SPSS统计软件对不同产地或不同采集时间的14批辛夷样品的指纹图谱数据进行聚类分析后,Ⅰ类中10批样品被选用来建立共有模式。以共有模式为参照,应用国家药典委员会《中药色谱指纹图谱相似度评价系统B版》分别计算各样品与共有模式之间的相似度。有两个批次的药材的相似度低于0.9,此结果与聚类分析结果一致,两种方法得到了相互验证,说明HS-SPME-GC指纹图谱可用于辛夷药材的质量评价。
同时,本文还首次建立了辛夷的HPLC指纹图谱。首先,在流动相的组成上,比较了不同流动相组成的分离效率,最终确定以水和乙腈作为流动相。其次在提取方法上,分别使用不同的提取溶剂,以提取效率为目标筛选,确定乙酸乙酯为最终提取溶剂。另外,在检测器方面,比较了蒸发光散射检测器和紫外检测器的差异。与蒸发光散射检测器相比,紫外检测器灵敏度高,出峰多。在优化的色谱条件下,各批辛夷药材的相似度均达到了0.9以上,符合指纹图谱的要求,为其资源品质的评价提供了更充分的科学依据。
The Magnolia Flos is our country's traditional traditional Chinese medicine, Plants the people's hopes for the wooden orchid aceae spring to spend Magnolia biondii Pamp., magnolia Magnolia denudata Desv. Or Magnolia sprengeri Pamp. dry flower bud. The winter end spring first flowering has not opened when recovers, except a stem, dries in the shade.
The Magnolia Flos simultaneously is also our province road place raw material for medicine, besides volatile oil, but also includes the wooden fat element, the alkaloid and so on each kind of effective component, has the sterilization, the voltage dropping and so on many kinds of pharmacological actions, although at present had other scholars to formulate Magnolia Flos raw material for medicine volatile oil GC and the GC-MS fingerprint, but limits in the extraction method to the traditional steam distillation, time-consuming is long, and is big to raw material for medicine and the resolver consumption, But regarding the Magnolia Flos in the other type's effective component, through establishes its highly effective liquid phase fingerprint atlas, between the research different technology obtained fingerprint atlas's relations, may let the fingerprint true reflection Magnolia Flos the intrinsic quality. In addition, in the literature also not yet sees the Magnolia Flos to go against the spatial solid phase micro extract- gas chromatography fingerprint and the highly effective liquid chromatography fingerprint report. Therefore the research raw material for medicine Magnolia Flos goes against the spatial solid phase micro extract-gas chromatography fingerprint atlas and the highly effective liquid chromatography fingerprint, provides the method and the basis for the Magnolia Flos raw material for medicine's distinction and the quality control, has the very vital significance.
This article established the Magnolia Flos to go against the spatial-solid phase micro extract-gas chromatography for the first time (HS-SPME-GC) the fingerprint, has provided one better method for system's science appraisal and the appraisal Magnolia Flos's quality. Using the HS-SPME technology directly extracted the volatile organic compounds in Magnoliae Flos, then these compounds be separated and identificated by GC and GC-MS. The Chromatographic fingerprint of Magnoliae Flos was established by HS-SPME-GC. The 47 common peaks were obtained and the 31 common peaks were displayed in the them by HS-SPME-GC-MS. The 14 samples were classified as 3 grades by cluster analysis and the 10 superior were confirmed to establish the mutual models, as a reference to it, the similarity were calculated between the samples and the mutual models, the results of cluster and similarity analysis remained the same. This research method for evaluating the quality of Magnoliae Flos provides scientific basis.
At the same time, this article also for the first time used the highly effective liquid phase chromatography (HPLC) to establish the Magnolia Flos Central Africa volatile constituent HPLC fingerprint, has provided the basis for the quality synthetic evaluation Magnolia Flos raw material for medicine's intrinsic quality. First, in the flowing composition, compared with the separation efficiency which the different flowing composed, determined finally took the flowing by the water and the methyl cyanide. Next in the extraction method, uses the different extraction resolver separately, take the extraction efficiency as goal screening, the definite ethyl acetate to withdraw the resolver finally. Moreover, in detector aspect, compared with evaporation light scattering detector and ultraviolet detector's difference. Compares with the evaporation light scattering detector, because the ultraviolet detector the sensitivity is higher than the peak to be many. Under the optimized chromatograph condition, each batch of Magnolia Flos raw material for medicine's similarity has achieved above 0.9, meets the fingerprint atlas requirement, has provided a fuller scientific basis for its resources quality's appraisal.
引文
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