Research On The Effective Constituents Of Chinese Honeylocust Spine And Quality Resources Of Forsythia Suspensa(Thunb.) Vahl From Chinese Herbal Medicine

To provide a scientific basis for the quality control of origin traditional Chinese medicinal materials, the key scientific objectives of this study were to isolate and identify the anticancer and antibacterial constituents from Chinese Honeylocust Spine and investigate the quality resources of Forsythia suspense(Thunb.) Vahl. The thesis thus contains two parts:(1) The separation of the effective constituents of Chinese Honeylocust Spine;(2) The study of the quality resources of Forsythia suspense(Thunb.) Vahl.Part 1: The separation, extraction and structural characterization of the effective constituents from Funiu mountain Chinese Honeylocust Spine. Chinese Honeylocust Spine, belonging to legumes, is the drying spines of Gleditsia sinensis Lam. As a Chinese traditional medicinal material with wide ranges of medicinal values, Chinese Honeylocust Spine has a long history of application in both clinics and folks. More recently, it raised a great attention in the academia. In the literature, it was widely reported that triterpenoid saponins and flavonoids are the medicinal compounds of Chinese Honeylocust Spine and mainly responsible for its anticancer and antibacterial activities. Using in vitro anticancer models, this work screened the new effective flavanones skeleton derivatives with low toxicity. Also antimicrobial tests revealed that sterol glycosides and flavonoids quercetin were the main effective constituents. In this study, nighteen compounds(belonging to four classes) were isolated from petroleum ether and ethyl acetate extracts of Chinese Honeylocust Spine using separation methods of Preparation of High Performance Liquid Chromatography(RP-HPLC), gel permeation chromatography(Sephadex HL-20) and silica gel column chromatography(CC). The molecular structures of the isolated compounds were identified and inferred by the analysis methods of Infrared Spectroscopy(IR) and Nuclear Magnetic Resonance Spectroscopy(NMR)(including two dimensional NMR). They are sterols and their glycosides, flavones and flavanones, coumarins and lignans, respectively. Compound 17 and compound 18 are novel compounds. The cholestane derivatives(compounds 3~5), flavanones cytoskeletal components(compounds 7~12) and coumarins and lignans derivatives(compounds 13~19) were isolated for the first time from the Chinese Honeylocust Spine. The current results offer the targeted material basis of the traditional medicinal uses of Chinese Honeylocust Spine for the first time.Part 2: Identification of the quality resources of traditional Chinese medicine A new technology of High performance liquid chromatogrpahy-electrospray ionizationtandern mass spectrometry(HPLC-ESI-MS) was employed to inveistigate the variation among its different species, thus providing the scientific basis for a comprehensive evaluation system of wild Forsythia resources. Six samples of different habitats and species of Forsythia suspensa(including Qingqiao and Laoqiao) were collected from three Chinese areas including Henan, Shanxi and Shaanxi. The quantities of effective constituents forsythiaside A, phillyrin and forsythiaside were then determined. The results showed that the Funiu mountain Qingqiao has the highest levels of effective compounds. The amount of effective constituents of Shanxi or Shaanxi Laoqiao is higher than that of Funiu mountain Laoqiao. The results above suggest that the doctor should select the prescription drugs based on scientific date. Compared to the HPLC analysis HPLC-MS method showed a faster and accurater performance in the quality control of Chinese herbal medicines. Moreover, it is simple, feasible and has a strong specificity in quantification analysis.