Studies On Constituents In Cultured Cells Of Saussurea Involucrata And A New Recycle Chromatography System

Saussurea involucrata is a well-known Chinese herbal medicine plant which is at the risk of extinction. As a potential resource for clinical usage, the cultured cells of S. involucrata (CCS) might theoretically generate comparable secondary metabolites. This dissertation studied chemistry of the CCS, and provide basic supporting information for the qulity control and further development of this new resource. In other hand, this study also developped a novel recycle productive chromatography system, which was further proved to be a powfer tool for separating a trace compound in complex sample.The first part of this study reviewed the constituents of most popular10ethnomedicinal plants of Xuelianhua, such as phenylpropanoids, coumarins, lignans, alkaloids, terpenes, flavonoids, steroids, and so on. And these would offer a good reference to understand the chemical composition in the CCS. Meanwhile, it was also reviewed of the research progresses in the fields of on-line sample treatments in chromatography and recycle chromatography, which provided important informance for developing a novel productive recycle HPLC system.In the second part of this study,41compounds were isolated and identified from cultured cells of Saussurea involucrata, which belonged to kinds of phenolic acid, flavone, lignin and so on. The results showed caffeoyl quinic acid type compounds were most abundant. And there contained novel and characteristic compounds of maloyl caffoeyl quinic acids, which were demonstrated to possess desire antioxidant activities in vitro anti-free radical experiments. Then it was developped a quantitative analysis method of five characteristic compounds (3-O-caffeoyl quinic acid,1,5-O-dicaffeoyl quinic acid, syringin, and two maloyl caffeoyl quinic acid type compounds) from the CCS extract. And these could contribute initial reference information for the further quality control and optimization of cultivative technology.The third part of this study pursued to develop a novel and efficient recycle preparative HPLC system. Compared with a standard preparative HPLC apparatus, in brief, this system contained an added trap unit which could serve as a multi-functional unit for sample-trapping, component-capturing, trace-enriching, recycle injecting and the final products concentrating, so as to separate low abundant components in complex sample with a high efficient method. Separation methods of the three new maloyl-caffeoyl-quinic acid type compounds were developped using this new chromatography system. Furthermore, polyphenol compounds were recycle-separated to pursue the polarity limity of samples for trapping, the result showed that the compounds with middle or low polarity (similar with or weaker than3-O-caffeoyl quinic acid) could be separated using this system. As an expend research, this new recycle HPLC system was used to separate a low abundant (<1%) and unstable impurity in crude pharmaceutical silybin. In all, it could be demonstrated this new recycle preparative HPLC system a powerful tool for separation, especially for purifying a low abundant compound from complex mixture in both natural products and pharmaceutical separation fields.