Analysis And Separation Of Chemical Constituents Of Apocynum Venetum And More Of Traditional Chinese Medicine

Chinese traditional medicinal herbs are abundant resources in the world. It made amost significant contribution for human health in the long history of Chinese nation, andalso do it today. Because the ingredients of traditional Chinese medicine are complex andthe contents of active ingredients are low and difficult to purify so the high purity activeingredients are essential to the medicine contents’ determination and pharmacologicalstudies as well as the control of medicine qualities. Therefore, it’s necessary to establish anefficient and effective method to separate and purify the ingredients of traditional Chinesemedicine.In this paper, the method of separation and purification the ingredients of traditionalChinese medicine has been studied, and then built a new separation method for theapocynum venetum and teasel root. In this experiment, the detection conditions, theproportion of the mobile phase and the flow rate and the injection volume were optimized.The target compounds were separated and purified by PHPLC and polyamide columnchromatography, and determined by HPLC. The obtained target compounds’ structureswere identified by UV and NMR.1. Isolation and Purification of the flavonoids from Apocynum venetum leafA new method for separation and purification of the flavonoids from the leaves of theapocynum venetum was established by PHPLC. Firstly, the extraction conditions wereoptimized. Secondly, crude extract after separation of the polyamide columnchromatography, obtained the flavonoids and oter compound. Finally, the preliminarypurification of the flavonoids were isolated by PHPLC. The best PHPLC chromatogaphicconditions were obtained when YMC ODS-A column (400mm×40mm I.D.,10μm) wasused at room temperature, the mobile phase was acetonitrile-water (18:82, v/v), theinjection volume was2mL, the flow rate of the mobile phase was20mL/min and thedetection wavelength was360nm. Finally, nine flavonoids were separated from leaves ofapocynum venetum. Nine compounds were achieve at last and they were elucidated bymeans of spectroscopy (UV,13C-NMR,1H-NMR).They were: quercetin-3-O-β-D-Glc(2 �'1)-β-D-Glucose(1)、 astragaloside(2)、(-)-gallocatechin（3）、rutin(4)、hypericin(5)、quercetin-3-O-β-D-Glucose(6)、kaempferol-3-O-β-D-Glucose(7)、quercetin-3-O-β-D-Glucose(8)、Isorhamnetin-3-O-β-D-Glucose(9).2. Isolation and Purification of the Iridoids and other compounds from the teaselrootsA new method for separation and purification of the iridoids and other compoundsfrom the teasel root was established by PHPLC. Firstly, the extraction conditions wereoptimized. Secondly, in order to separate the iridoids from other compounds, the crudeextracts were extracted by light petroleum and ethylacetate. At last, the purification of theiridoids were isolated by PHPLC. It was performed on a C18column (420mm×40mmI.D.,50μm) at room temperature. Thirdly, the preliminary purification of the iridoids wereisolated by PHPLC. The best PHPLC chromatogaphic conditions were obtained whenYMC ODS-A column (400mm×40mm I.D.,10μm) was used at room temperature, themobile phase was acetonitrile-water (20:80, v/v), the injection volume was2mL, the flowrate of the mobile phase was20mL/min and the detection wavelength was210nm.Finally, three iridoids were separated from the teasel roots. Three compounds were achieveat last and they were elucidated by means of spectroscopy (UV,13C-NMR,1H-NMR).They were:Epi-vogeloside、vogeloside andβ-sitosterol.