【作者】 徐小娜；

【导师】 梁逸曾；

【作者基本信息】 中南大学 ， 分析化学， 2010， 博士

【摘要】 当前的中药现代化是在信息化时代,科学技术高度发展,人们重新崇尚回归自然及我国对外开放,面向全球的大环境下的又一次新的浪潮。尽管中药所含物质群非常复杂,但联用仪器和相关化学计量学方法的快速发展为中药这一复杂体系的分析与质量控制提供了强有力的手段和广阔的前景。近年来,指纹图谱技术在国际上已广泛达成共识,色谱指纹图谱质量控制是中药质量控制领域的热点和趋势。本论文以实际中药体系为研究对象,利用HPLC-DAD和GC-MS联用技术,结合相关化学计量学方法,进行中药色谱指纹图谱研究,为中药质量控制及药理、药效研究提供基础数据和理论依据。论文的第一部分即第一章,论述了近年来我国中药质量控制模式的发展状况,中药指纹图谱,现代分析技术以及化学计量学解析分辨技术和模式识别方法在中药质量控制中的应用等内容。第二部分为HPLC-DAD联用技术和相关化学计量学方法用于中药枳实色谱指纹图谱的研究(第二章和第三章)。其中,第二章详细地阐述了枳实中黄酮类成分的高效液相指纹图谱研究。内容主要包括样品提取条件和色谱分离条件的优化；成分分析,尤其是活性化合物的定性定量分析；首次将直观推导式演进特征投影法(heuristic evolving latent projections, HELP)用于枳实二维液相色谱指纹图谱中重叠色谱峰的解析；相似度比较分析法(Similarity comparative analysis,SCA)用于12批枳实的分析等。第三章在第二章的基础上将化学模式识别方法——主成分分析法(Principal component analysis, PCA)和聚类分析法(Hierarchical clustering analysis, HCA)用于不同来源的38个枳实样本的色谱指纹图谱研究,对两类样本的指纹图谱的共有模式及可疑样本的指纹图谱进行比较分析,对活性化合物进行定量分析。发现,中药(材)由于来源复杂(如产地)、采集时间、炮制方法、储存条件、储存年限等不同,使得其所含有的化学物质在种类和含量上(特别是活性化合物的含量)存在较大差异,建议有关部门应趁早采取措施,规范药材中那些已明确药理作用的活性化合物的测定方法和含量指标；对中药材进行分类研究时,以完整的色谱指纹图谱数据做输入变量比以少数主要色谱峰的峰面积做输入变量更能彰显样本所包含的化学物质的全部信息及发现可疑样本。研究表明,将化学计量学方法与中药指纹图谱研究相结合是科学的、全面的,有利于中药质量控制。第三部分为三种化学计量学分辨方法用于药对紫苏-藿香挥发油成分的分析(第四章)。在这一部分中,详细地论述了三种化学计量学分辨方法——交替移动窗口因子法(Alternative moving window factor analysis, AMWFA)、直观推导式演进特征投影法(HELP)和选择性离子分析法(Selective ion analysis, SIA);首次同时运用上述三种分辨方法解析药对紫苏-藿香及其单味药挥发油成分的GC-MS数据；此外,计算程序升温保留指数(Temperature-programmed retention indices, PTRIs),用来辅助化合物的定性分析。所获结果表明,综合运用此三种化学计量学分辨方法能比较全面地对中药的挥发油成分进行解析,鉴定出更多的组份,使定性定量分析更准确。比较分析药对及其单味药的挥发油成分,发现药对紫苏-藿香的挥发油成分的总数目略小于两个单味药的挥发油成分的数目之和,药对挥发性成分中的主要成分均来自两单味药。所得结果为探索该药对的药理作用及其与单味药的化学成分的关系提供理论基础,为复方中药研究提供新思路。第四部分为直观推导式演进特征投影法(HELP)用于不同药用部位的厚朴挥发油成分的分析(第五章)。首次利用HELP法对湖南产凹叶厚朴的不同药用部位(干皮、枝皮和根皮)的挥发油成分的GC-MS数据进行解析。同时,计算程序升温保留指数(PTRIs),提高定性分析的可靠性。结果：从厚朴干皮、枝皮和根皮的挥发油成分中分别定性出90,82和76种化合物,三者共有组分50个；首次在该药材的挥发油成分中同时定性、(半)定量分析了桉油醇的三种同分异体(α-,β-和γ-)；比较分析表明,厚朴干皮、根皮和枝皮的挥发油成分存在一定的相似性和差异性。本部分内容为厚朴药材资源的合理使用及药理药效研究提供参考。第五部分为直观推导式演进特征投影法(HELP)和交互移动窗口因子分析法(AMWFA)用于化湿药挥发油成分的分析(第六章)。运用HELP法和AMWFA法,首次对五种常用化湿药(砂仁、苍术、藿香、佩兰和厚朴)的挥发油成分的GC-MS数据进行比较分析,利用程序升温保留指数(PTRIs)辅助定性。分析五种化湿药挥发油成分中的共有组分、主要成分及活性成分,比较分析它们挥发油成分的异同,对化湿药进行释义,为深入该类中药的药理、药效研究和诠释中药的分类打下基础。本论文将二维色谱联用技术和相关化学计量学方法与中药色谱指纹图谱研究相结合,对中药枳实、药对紫苏-藿香、厚朴及常用五种化湿药进行了一些基础研究,充分展现了这种结合在中药研究方面的优势和潜力。更多还原

【Abstract】 The modernization of traditional Chinese medicine (TCM) is a new tidal wave in the era with informationization, highly developed science and technology. Nowadays, people want to go back to the nature again. Although the components of TCM are extremely complex, the rapid development of modern hyphenated instruments and related chemometric methods make it possible to analyze TCM system and control its quality. The technology of chromatographic fingerprint is widely accepted by international society, and the quality control by chromatographic fingerprint is the hot spot and the trend in these years. So, this thesis aims at the practical TCM system, applys chemometric methods to the analysis of group substances in real TCM system in order to control its quality and provide useful information for the research of herbal medicine.The first fraction of this thesis is the first chapter. In this fraction, the development of TCM quality control in recent years, the application of chromatographic fingerprints, the progress of modern analytical techniques and chemometric methods are summarized.The second fraction of this thesis emphasizes on the applications of high performance liquid chromatography-diode array detector (HPLC-DAD) and correlative chemometic methods to the research of the components and chemical fingerprint analysis of Fructus Aurantii Immaturus (FAI) (chapter 2 and chapter 3). In chapter 2, the optimization of extraction and chromatographic separation conditions to establish a standard chromatographic fingerprint of FAI, and the analysis of the components and especially the bioactive compounds were discussed in detail. Meanwhile, heuristic evolving latent projection (HELP) method was employed to resolve the two-dimension data for the first time. Based on the results of Chapter 2, Chapter 3 aims at the employment of chemical partten recognition methods, say principal component analysis (PCA) and hierarchical clustering analysis (HCA), to the analysis of the 38 FAI fingprints from the entensive origins of China. The results obtained show that masses of factors such as the various resources, collection time, processing methods, storage conditions and so on, have a big influence on the contents of chemical compounds of TCM, which indicates the correlative departments to formulate test methods and content standards of those compositions, especially the bioactive ones; in the research of classification of TCM, the entire fingerprint data has strong ability to display the whole information of TCM samples than the partial data of the fingerprints, which helps to find doubtful samples. The results obtained show that the combination of chemometric methods and fingerprint technique is sciencfic and allround, which can be applied to quality control of TCM.The third fraction of this thesis discusses the application of three chemometric resolution methods to the analysis of the volatile oil compositions of drug pair (DP) P. cablin-P. frutescens (chapter 4). In this fraction, three chemometric resolution methods, namely alternative moving window factor analysis (AMWFA), HELP and selective ion analysis (SIA), were expounded and compared in particular. Meanwhile, the three methods forementioned were firstly employed to analyze the evaporable constituents of DP P. cablin-P. frutescens and its single herbs, which were detected by gas chromatography-mass spectrometry (GC-MS). In addition, temperature-programmed retention indices (PTRIs) were also used together with mass spectra for tentative identification of the essential oil constituents. The achieved results show the general use of multiple chemometric resolution methods display strong advantage in anlyzing the overlapped and embedded chromatographic peaks, and realizing fast comparative analysis between two complex systems. The comparative analysis between the DP and its single herbs shows that the number of essential components of the DP is a little less than the sum of the number of its two single herbs. The major components of the volatile oil of the DP are basicly from those of the two single herbs. The results obtained may provide a useful chemical basis for future research on the correlation between the pharmacodynamic action and chemical constituents of the DP and its single herbs, and benefit the further research of compound recipe.The fourth fraction aims at using the HELP resolution method in combination with GC-MS technique to the anlysis of Cortex magnolia officinalis (Chinese name ’houpo’) (chapter 5). In this fraction, volatile compositions of different parts (stem, branch and root barks) of houpo, cultivated in China, were investigated for the first time by GC-MS in combination with HELP method. Identification of components was conducted by similarity matching to NIST mass library but also assisted by comparison of PTRIs with the data web available. A total of 90,82 and 76 volatile compounds in the essential oils of the three samples taken from the same batch forementioned were qualitatively and quantitatively determined. Among the constituents determined, there were 50 components coexisting. The three kinds of isomers (α-,β- andγ-eudesmol) in houpo essential oil have been tentatively qualified and quantified simultaneously for the first time. Significant qualitative and semi-quantitative differences and similarities were observed among the three samples through comparative analysis. The results achieved provide a scientific evidence for further exploitation of Magnolia bark and clinical medication.The fifth fraction employs chemometric methods and GC-MS technique to the analysis of five aromatic drugs for resolving dampness (chapter 6). In this fraction, comparative analysis of essential oil ingredients from Fructrs Amomi, Rhizoma Atracylodis, P. cablin, Herba Eupatorii and houpo, was firstly carried out by two chemometric methods, say AMWFA and HELP. The PTRIs were also used to aid qualitative analysis. The common constituents of the five herbs forementioned were analyzed, together with the principal compositions and the bioacitve ones. By comparison of the differences and similarities of their chemical components, the reason why they are classified into the dampness-dissipating herb medicine was explained to some extent. The results obtained will be good for further research of pharmacological action and annotation of classification of TCM.In this thesis, multi-dimensional chromatography and correlative chemometric methods were applied to the study of constituents and chromatographic fingerprints of TCM, showing their advantage and huge potential in the research of TCM. Some fundamental researches about FAI, DP P. cablin- P. frutescens, houpo and five aromatic drugs for resolving dampness were done, which will benefit the resarch of TCM.更多还原