龙胆泻肝丸等三种中药复方的质量控制研究
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摘要
中药复方是在中医理论指导下,按照“君臣佐使”的组方原则,选择适宜的药味和剂量配伍而成的一类传统制剂。中药复方成分复杂,长期以来化学物质基础研究的匮乏成为制约中药复方发展的重要因素,影响了中药现代化和国际化征程,因此有必要综合现代分析方法对中药复方进行物质基础研究,并在此基础上建立中药复方质量控制方法,以保证中药质量控制的合理性和有效性。
本课题以传统中药复方龙胆泻肝丸、附子理中丸和乌鸡白凤丸作为研究对象,依据中药现代化质量控制的思想,对其物质基础进行了系统研究,建立了指纹图谱和多成分含量测定方法；并采用系统聚类分析法对不同厂家的样品指纹图谱进行聚类分析；此外还利用液质联用和气质联用技术进行化学成分定性研究,为物质基础和质量控制研究提供了稳定可靠的分析条件。结论如下：
11采用HPLC-DAD法建立了龙胆泻肝丸的化学指纹图谱,以及采用HPLC-MS法对龙胆泻肝丸中12个含量较高的化合物进行了定性分析,明确其化学结构,阐明了龙胆泻肝丸的化学物质基础,同时测定该12个活性成分的含量,并采用系统聚类分析法对不同批次的龙胆泻肝丸样品进行了聚类分析。另外,对该方法进行了系统的方法学考察,结果表明该方法适合于龙胆泻肝丸的质量控制研究和化学物质基础表征。
21首次采用RRLC-Q-TOF及GC-MS法建立了附子理中丸的化学指纹图谱,对附子理中丸中45个色谱峰进行了定性分析,明确了化学结构,阐明了附子理中丸的化学物质基础。依据RRLC-Q-TOF指纹图谱,并采用系统聚类分析法对来源于多个不同厂家的附子理中丸样品进行了聚类分析,结果表明,不同厂家的附子理中丸存在较大差异。
31采用RRLC-Q-TOF法建立了乌鸡白凤丸的化学指纹图谱,对乌鸡白凤丸中的10个色谱峰进行了定性分析,明确了其化学结构,阐明了乌鸡白凤丸的化学物质基础。依据RRLC-Q-TOF指纹图谱,并采用系统聚类分析法针对来源于不同厂家的乌鸡白凤丸样品进行了聚类分析,结果表明,不同厂家的乌鸡白凤丸之间的差异较小。
In this study, three Traditional Chinese Medicines (TCMs), Longdan Xiegan Pill (LXP), Fuzi Lizhong Pill (FLP) and Wuji Baifeng Pill (WBP), are selected as the object of research. The quality evaluation system was systematically investigated, which is on the basic of modem quality control trend. The method was developed on the chromatographic fingerprints and simultaneous quantification of the major components of TCMs. In addition, the fingerprints were qualitatively evaluated by hierarchical cluster analysis. This method set a foundation for quality control of TCMs. The results were summarized as follows:
1) A HPLC coupled with DAD and MS spectrometry method was developed for the quality assessment of LXP by combination of chromatographic fingerprint and simultaneous quantification of 12 major components. These compounds were identified by ESI/MS. In addition, the samples according to their manufacture corporation were classified, and 12 major ingredients were screened out to be the most discriminant variables which could be regarded as the constituents for quality assessment of LXP by using Hierarchical Cluster Analysis. The proposed method was utilized for the quality control and original discrimination of LXP successfully.
2) A RRLC-Q-TOF and GC-MS method was developed for the quality assessment of FLP by combination of chromatographic fingerprint. We identified 45 components in FLP. In addition, based on the fingerprint data, (HCA) was applied in qualitative evaluation on the difference and resemblance of FLP.
3) A RRLC-Q-TOF method was developed for the quality assessment of WBP by combination of chromatographic fingerprint. We identified 10 components in WBP. Then based on the fingerprint data, hierarchical cluster analysis (HCA) was applied in qualitative evaluation on the difference and resemblance of WBP.
本课题以传统中药复方龙胆泻肝丸、附子理中丸和乌鸡白凤丸作为研究对象,依据中药现代化质量控制的思想,对其物质基础进行了系统研究,建立了指纹图谱和多成分含量测定方法；并采用系统聚类分析法对不同厂家的样品指纹图谱进行聚类分析；此外还利用液质联用和气质联用技术进行化学成分定性研究,为物质基础和质量控制研究提供了稳定可靠的分析条件。结论如下：
11采用HPLC-DAD法建立了龙胆泻肝丸的化学指纹图谱,以及采用HPLC-MS法对龙胆泻肝丸中12个含量较高的化合物进行了定性分析,明确其化学结构,阐明了龙胆泻肝丸的化学物质基础,同时测定该12个活性成分的含量,并采用系统聚类分析法对不同批次的龙胆泻肝丸样品进行了聚类分析。另外,对该方法进行了系统的方法学考察,结果表明该方法适合于龙胆泻肝丸的质量控制研究和化学物质基础表征。
21首次采用RRLC-Q-TOF及GC-MS法建立了附子理中丸的化学指纹图谱,对附子理中丸中45个色谱峰进行了定性分析,明确了化学结构,阐明了附子理中丸的化学物质基础。依据RRLC-Q-TOF指纹图谱,并采用系统聚类分析法对来源于多个不同厂家的附子理中丸样品进行了聚类分析,结果表明,不同厂家的附子理中丸存在较大差异。
31采用RRLC-Q-TOF法建立了乌鸡白凤丸的化学指纹图谱,对乌鸡白凤丸中的10个色谱峰进行了定性分析,明确了其化学结构,阐明了乌鸡白凤丸的化学物质基础。依据RRLC-Q-TOF指纹图谱,并采用系统聚类分析法针对来源于不同厂家的乌鸡白凤丸样品进行了聚类分析,结果表明,不同厂家的乌鸡白凤丸之间的差异较小。
In this study, three Traditional Chinese Medicines (TCMs), Longdan Xiegan Pill (LXP), Fuzi Lizhong Pill (FLP) and Wuji Baifeng Pill (WBP), are selected as the object of research. The quality evaluation system was systematically investigated, which is on the basic of modem quality control trend. The method was developed on the chromatographic fingerprints and simultaneous quantification of the major components of TCMs. In addition, the fingerprints were qualitatively evaluated by hierarchical cluster analysis. This method set a foundation for quality control of TCMs. The results were summarized as follows:
1) A HPLC coupled with DAD and MS spectrometry method was developed for the quality assessment of LXP by combination of chromatographic fingerprint and simultaneous quantification of 12 major components. These compounds were identified by ESI/MS. In addition, the samples according to their manufacture corporation were classified, and 12 major ingredients were screened out to be the most discriminant variables which could be regarded as the constituents for quality assessment of LXP by using Hierarchical Cluster Analysis. The proposed method was utilized for the quality control and original discrimination of LXP successfully.
2) A RRLC-Q-TOF and GC-MS method was developed for the quality assessment of FLP by combination of chromatographic fingerprint. We identified 45 components in FLP. In addition, based on the fingerprint data, (HCA) was applied in qualitative evaluation on the difference and resemblance of FLP.
3) A RRLC-Q-TOF method was developed for the quality assessment of WBP by combination of chromatographic fingerprint. We identified 10 components in WBP. Then based on the fingerprint data, hierarchical cluster analysis (HCA) was applied in qualitative evaluation on the difference and resemblance of WBP.
引文
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