Study On Active Components And Quantitative Analysis Of Selaginella Doederleinii Hieron

It is well known that cancer is a kind of disease causes seriously damage to humanlife and health. Chinese herbal medicines have been widely accepted by theinternational society in recent years with its significant clinical effect in anti-tumorand low adverse reactions advantages. Thus, currently, the research and exploitationof anti-tumor Chinese herbal medicine has become a hot topic.Selaginella doederleinii Hieron belongs to the Selaginellaceae Genus plants ofPteridophyta. It is widely used to treat sore throats, rheumatic arthralgia pain,nasopharyngeal carcinoma, chorionic carcinoma, lung cancer, cervical cancer and avariety of diseases such as traumatic bleeding. However, its anti-tumor effect of thematerial basis has reported relatively less. Therefore, it is necessary to conductin-depth study in the aspect.In this dissertation, the primary extraction and separation of S. doederleinii wascarried out, and its ethanol extract and different polar organic solvent extracts werefurther screened in vitro for anti-tumor activity; subsequently, the activity extractsscreened out were suffered with further separation procedures by preparative liquidchromatography for obtaining the main active ingredients and the structures of theobtained compound were identified by NMR and MS, etc; the content determinationmethod of the main components was also developed. In addition, taking as a marker,the pharmacokinetic behavior of S. doederleinii extract in rat plasma was investigated.The main contents and achievements are as follows:1. Preliminary extraction and separation of S. doederleinii. Ethanol total extract wasobtained with70%ethanol as the extraction solvent, followed by extraction in turnwith petroleum ether, methylene chloride and ethyl acetate as the extraction solvents.The organic solvent extracts and the residual water portion were finally concentratedto give the corresponding extracts. Then these extracts were suffered with MTTmethod with human cancer cells to assess their anti-tumor activities in vitro. As aresult, the ethyl acetate extract shows good anti-tumor activity in vitro. 2. The separation and purification of ethyl acetate extract was carried out by usingthe preparation liquid chromatography. Four compounds were successfully obtainedfrom this extact. Their structures were identified as amentoflavone(I),robustaflavone(II),2",3"-dihydro-3’，3’"-biapigenin(III) and3’,3"’-binaringenin(IV)by NMR and MS techniques. Among them, compounds III and IV were firstlyisolated from this plant.3. The HPLC method for the determination of the major four constituents,amentoflavone, robustaflavone,2",3"-dihydro-3’，3’"-biapigenin and3’,3"’–binaringenin in S. doederleinii was developed. The HPLC method established wassuccessfully used for the quantitation analysis of this medicine herb.4. The HPLC-MS method for determination of amentoflavone in rat plasma wasdeveloped. With the developed method, the pharmacokinetics of amentoflavone afteroral administration of S. doederleinii extract in rat was investigated. This study ishelpful to reveal the process of S. doederleinii extract in vivo and the material base ofpharmacological function.