Hyphenated Chromatography Applied To Traditional Chinese Medicine And Chemical Medicine Preparation For Their Quality Control

It is the basic principle for drug development and evaluation to guarantee the safety, efficiency and quality-control for medicines, in which quality control of the medicines is the basis and precondition. Recently, Chinese and natural medicines attract more and more attention all over the world. Although Chinese and natural medicines are very complex, hyphenated instruments, such assuch as GC-MS, HPLC-DAD, HPLC-MS, combined with chemometrics provide strong tools for their analysis and quality control. On the other hand, the inpurity control becomes a very imortant concern for chemical medicine preparation. With the increasing understanding of the inpurities in chemical medicine preparation, the concept of "control of inpurity spectrum" is widely accepted all over the world. This thesis just focuses the application of hyphenated instruments combined with chemometrics for quality control of traditional Chinese medicine and chemical medicine preparation.The content of the thesis is composed ofthe following sections:1. The composition analysis of Codonopsis pilosula collected from different regions using twochemometric resolution methods is firstly investigated.(Chapter2) In this chapter, we firstly discussed the two chemometric resolution methods, say Heuristic Evolving Latent Projection Method (HELP) and Selective ion analysis (SIA), in details. Using both the above two resolution methods, the GC-MS data of volatile constituents in Codonopsis pilosula from different areas were analyzed. In addition, temperature-programmed retention indices (PTRIs) are introduced for qualitative analysis of the active compounds, in order to improve the reliability of qualitative analysis. The results obtained show that volatile oil components in the Codonopsis from Gansu, Shanxi and Xinjiang are69,78and63, respectively. The content of qualitative components accounted for the total content of volatile oil were89.52%,91.65%and90.35%, respectively. There are29common volatile oilsin Codonopsis pilosula from three different regions. The comparative analysis shows that, differences and similarities exist in the volatile oil components of Codonopsis from different regions.This section provides reference for rational use of Codonopsis resources.2. Taking volatile components of Traditional Chinese medicine SHXW as the object of the study, a simple and facile GC-MS fingerprint of SHXW was firstly developed. Then, the similarity analysis was conducted. However, a neglected problem was discovered on certain important components in the share of fingerprints. The problem is that the degree of similarity is sometimes very high (more than95%) even without a certain ingredient in SHXW. That means it is very difficult to fully address the quality control of SHXW by simply using the degree of similarity between the chromatographic fingerprints. It seems necessary to further research on how to establish the novel criterion of fingerprinting for quality control of TCMs. Firstly, with the help of the temperature-programmed retention indices (PTRIs) used together with mass spectra and chemical standards,25major characteristic peaks have been identified. Then, comparisons among references of Chinese herbal medicines, between the full preparation and negative controls of preparation were carefully conducted focusing on the identification and assessment of the attribution of the main characteristic peaks. Nine volatile medicinal materials were identified in the prescription of SHXW by attributing to the27major characteristic peaks. The results demonstrated that the proposed method was a powerful approach for quality control of complex prescription of herbal medicines.4. Research on different species of mercuryin SHXW (chapter4). By using the method of HPLC-AFSanalysis of different mercury species of cinnabar-containing traditional Chinese medicine SHXW, analysis of results of concentrations of Hg2+in SHXW and cinnabar in acid solution (pH=1.3), and in simulate gastric acid, it was found that the MeHg and EtHg were not detected. Moreover, the concentrations of Hg2+in liver and kidney of rat after giving SHXW for10days, the results demonstrated that the mercury species of SHXW and cinnabar in acid solution and in simulate gastric acid were all soluble Hg2+without the MeHg and EtHg. The results show that the SHXW is much less toxic than MeHg and EtHg. The results obtaned from this work interpreted the mercury poisoning of cinnabar preparations, which will be useful for explaining the foundation for the pharmacology and toxicity of this kind of traditional Chinese Medicine. Through re-evalution for cinnabar-containing traditional Chinese medicine SHXW, this will especially lay the foundation of revising the quality standard of safety of cinnabar in Chinese Pharmacopoeia. The researches on pesticide residues in SHXW and the corresponding medicinal herbs in SHXW were investigated in detail in chapter5of the thesis.5. In order to guarantee the safety of clinical drugs, the re-evaluation of technology and impurity profile for ribavirin injection were investigated in some detail (chapter6). With the help of HPLC-MS, the impurity profile of the ribavirin preparation was established.Through the analysis of the change of impurity profiles of the ribavirin preparationand the correspnding raw material by acid, base, light, heat and oxidation,the possible sources of theimpurities were confirmed in the production process. The main impurities were then analyzed qualitvely and quantitatively. According to the European pharmacopeia, several impurities, which are not limited before in Chinese pharmacopeia, were limited quantitatively for the first time.