Studies On The Chemical Constituents Of Anoectochilus Roxburghii

In this paper, the selection of the factors for extracting flavonoids compound, and detecting of the content of them were studied. We also developed a HSCCC method for the separation and purification of effective ingredient from Anoectochilus roxburghii. In these experiments, several methods such as UV-vis, IR, HPLC and NMR were used to identified the structures of chemical components which were separated by HSCCC.This thesis includes four parts as follows:PartⅠ: Quantitative analysis and Ultrasound-microwave assisted extraction of the flavonoids from A. roxburghii.The optimized condition of extraction of the sample studied with orthogonal design in ultrasound-microwave combined assay. To established a HPLC method for determination of quercetin, kaempferol and isorhamnetin in A. roxburghii. The reversed phase HPLC system consisting of an Agilent ODS column(4.6mm×250mm, 5μm) and a mixture of CH3OH-H2O-H3PO4 as the mobile phase was used. The detection wavelength was at 368 nm and the flow rate was at 1.0 mL·min-1.PartⅡ:Preparative isolation and purification of flavonoids compounds from A. roxburghii by HSCCC.HSCCC was successfully used for the isolation and purification of quercetin from A. roxburghii, The Chinese herbal drug, n-hexane-ethyl acetate–methanol–water (4:6:3:3, v/v) was used as the two-phase solvent system. 15.5 mg of quercetin with the purity of 96.5% was obtained from 2.8g of crude extracts of A. roxburghii. The structures of the isolated compounds were identified by UV-vis, IR, HPLC and 1H-NMR.PartⅢ:Preparative isolation and purification of sterins compounds from A. roxburghii by HSCCC.HSCCC was successfully used for the separation and purification of Ergosterol and Stigmasterol from the extracts of A. roxburghii. From 2.5g of the extracts, 36.5mg of Ergosterol and 43.6mg of Stigmasterol were obtained with purity of 92.0% and 95.5%. HSCCC condition: two-phase solvent syetem: n-hexane-ethylacetate- butanol- methanol-water (3.5:0.3:0.5:2.5:0.3,v/v). The chemical structures of these components were identified by UV-vis, IR, HPLC, 1H-NMR and 13C-NMR.PartⅣ: A review of the progress about hight-speed counter current chromatography was given.