| The constituent of traditional Chinese medicine (TCM) is multitudinous and complex. And also, its quality has been influenced by many factors, such as species, origin, harvesting time, processing of medicinal herb and the processing of preparation, which make its quality control become a "bottleneck" to restrict its industrialization and modernization. The assaying of active constituent is the key to its quality control. For Pharmacopoeia of The People’s Republic of China (Version2010, Part one), the idea of quality control has been changed from determination of single component to fingerprint and determination of multi-component. And also, thin-layer chromatography (TLC) and high performance liquid chromatography (HPLC) have been widely used for identification and determination of TCM. In this thesis, chromatographic fingerprint and multi-component determination were employed, and several new liquid chromatographic methods have been developed to determine main active constituents by TLC and HPLC in four TCMs and their praparations. Details of research are as follows:1. A HPLC-quantitative analysis of multi-components by a single marker (QAMS) method with a reference of podophyllotoxin was established to separate and determine four podophyllum lignans. The method was simple, accurate and reliable to control the quality of Dysosma versipellis (Hance) M. Cheng and Podophyllum emodi Wall. Var. chinesis Sprague; intra-class coefficient was used to evaluate the quantitative results of QAMS and the results of external standard method, and the results indicated that the developed QAMS method was accurate and reliable.2. A fingerprint of Hellebours thibetanus Franch was developed by HPLC-diode array detector coupled with hierarchical clustering and principal component statistical analysis. The method was used to evaluate17raw samples and1processed sample of Hellebours thibetanus Franch from4areas. The results showed that the degree of similarity from same area was good, and the similarity from different area was slightly different. And also, the raw and the processed samples could be also distinguished. The developed method has high repeatability and good stability and it could be useful for the authentication and quality control of H. thibetanus Franch.3. A hydrophilic interaction liquid chromatographic method was developed for simultaneous quantitative analysis of methylephedrine, ephedrine and pseudoephedrine in Ephedra Herb and its three preparations. The results of validation indicated that the developed method was rapid (in8minutes), accurate and sensitive for determination of the three ephedrine alkaloids in Ephedra Herb and its three preparations. According to the results of optimized chromatographic conditions, we have proposed a possible separation mechanism mode.4. A two-dimensional thin-layer chromatographic fingerprint was developed on a polyamide plate and using a non-aqueous mobile phase in the first dimension and a reversed micellar mobile phase in the second dimension, which provided a reference method to identify and quality control of Helleborus thibetanus Franch. Canonical correlation analysis was firstly used to determine the degree of similarity of thin-layer chromatograms. The effects of stationary and mobile phase on separation were studied systematically and on the basis of them the possible separation mechanism was discussed. |
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