Chemical Composition And Activity Analysis Of Extractive From Euscaphis Japonica Var. Wupingensis

Euscaphis japonica(E.japonica)var.wupingensis,a new variety of Euscaphis japonica(Thunberg)Kanitz,which belongs to genus of Euscaphis,a family of Staphyleaceae,has been widely cultivated in Fujian province.Its efficacy of pericarp included relieving swelling and pain,and centering in regulating the flow of vital energy.E.japonica consists of several chemical components i.e.phenylpropanoids,phenolic acids,flavonoids,alkaloids,aromatic compounds and triterpenoids possesses a variety of pharmacological actions including anti-inflammatory,anti-oxidation,anti-cell proliferation,anti-fibrosis,etc.The objective of this study was to investigate the chemical component of the husk and xylem extract from E.japonica.Meanwhile,the biological activity of the isolated compounds was also investigated.The results of this work would provide insights into the fundamental theory for the high effective utilization of E.japonica.,The main results are as follows:1.An online two-dimensional chromatographic analysis method was established to analyze the chemical structure of methanol extracts from E.japonica var.wupingensis and E.japonica.The results showed that the RPLC�RPLC two-dimensional chromatogram of E,japonica var.wupingensis and its varietes exhibited analogically.,only content difference existedin the overwhelming component with the exception of a distinct component,which has been verified by the central cutting analysis method This finding provides a theoretical basis for the separation of the chemical constituents extracted from the pericarp of E.japonica var.wupingensis coupled with wood species identification.2.An online antioxidant activity(HPLC-FRAP)device has been established testing the methanol extractives(MEs)composition in the pericarp and xylem of E.japonica var.wupingensis,the results revealed the methanol extractives in pericarp and the ethyl acetate extracts in xylem can be selected as separation and purification materials.3.The MEs in pericarp and EAs in xylem have been purified to obtain 31 compounds by using several techniques and online two-dimensional preparation.Total 30 compounds have been identified by physicochemical constant measurement,nuclear magnetic resonance and mass spectrometry.Among them,compoundseugennin,Quercetin-3-O-?-L-pyraninarabinoside,3,3'-di-O-methylalanine-4-(5"-ace tyl)-?-L-arabinofuranose,Boehmenan B,threo-Carolignan H,carolignan F were isolated from the genus for the first time.4.A DPPH free radical scavenging test was conducted and the results indicated that various compounds i.e.the MEs in the pericarp,flavonoids existed in ethyl acetate extracts(EAs)of xylem,phenylpropanoids and quinones represented differential scavenging activity on DPPH free radicals,of which the IC50 value of ellagic acid was 4.05?M and twice as the oxidant VC.5.The toxicity activity analysis of extracted compounds in E.japonica var.wupingensis on zebrafish showed that the triterpenoids and lignan compounds had a minor effect on the safety of zebrafish.However,both of the Tannin compounds and flavonoids can result in a severe effect on zebrafish safety,such as early egg coagulation death,late abdominal swelling and spinal deformity.Using the MTT method,the study on activity of isolated compounds was carried out by employing human hepatoma cells(HepG-2)cells as to inhibit cell proliferation as the detection target.The results showed that the lignans in the xylem of the yellow pheasant has inhibitory effects on the proliferation of HepG2 hepatoma cells,of which compounds 27,28 and 29 performed satisfactory inhibitory effects.The IC50 values of compounds 27,28 and 29 were 24.21 ?M,21.76 ?M and 41.79 ?M respectively..6.An HPLC method for the determination of the ketone compounds of the yellow fruit frog has been established.Both the content of isobiflorin and biflorin existed in the xylem,branch,pericarp of E.japonica var.wupingensis as well as pericarp of E.japonica,The results revealed that the content of chromones in a different location of the same plant varied sharply.Meanwhile,the isobiflorin content in the pericarp of E.japonica var.wupingensis(630.16?g/g)was more than that in E.japonica(576.87?g/g).In addition,the biflorin content of them varied minor.In summary,this thesis has carried out in-depth research on E.japonica var.wupingensis from chemical composition,biological activity and quality analysis,and provides theoretical guidance for the pharmaceutical application and development of E.japonica var.wupingensi.