Study On Chemical Constituents Of Clematis Huchouensis Tamura And Preparation Of Cream

Members of the genus Clematis,belonging to the family Ranurculaceae.There are more than 300 species in all,with about 110 distributed throughout China.Commonly used medicines include C.huchouensis,C.florida,and C.montana.C.huchouensis is included in the "National Chinese Herbal Medicine Compilation",often used as a whole herb,with anti-cancer,detoxification and swelling effects;external use can cure brain tumors and rheumatism arthritis.Objective:1.To study the chemical constituents in the ethyl acetate and n-butanol extracted for the 95% ethanol extraction of C.huchouensis.To find out more effective components in C.huchouensis,in order to rationally develop and apply C.huchouensis.2.The best water extraction technology of C.huchouensis was screened,and the cream was prepared by using the water extract of C.huchouensis as raw material,and the preliminary quality evaluation was carried out to provide reference for later clinical application.Methods:1.Separation and purification of ethyl acetate and n-butanol extracts from 95%ethanol extract of C.huchouensis by a variety of chromatographic separation techniques(Si CC,Sephadex LH-20,etc.).Various spectroscopy techniques(NMR,MS)and literature were used to analyze the monomers.The compound was subjected to chemical structure identification.2.The HPLC method for determination of oleanolic acid in C.huchouensis was established.The optimal water extraction process of C.huchouensis was optimized by orthogonal design method.The orthogonal design method was used to optimize the extraction process of oleanolic acid and yield rate.In the C.huchouensis water extract,the appropriate matrix and auxiliary materials were added to prepare C.huchouensis cream.Results:14 monomer compounds were isolated and identified from the ethyl acetate and n-butanol extracted for the 95% ethanol extraction of C.huchouensis,namely:physcion(1),methyl shikimate(2),kaempferol(3),ferulic acid(4),oleanolic acid(5),dibutyl phthalate(6),diisobutyl phthalate(7),β-sitosterol(8),stigmasterol(9),kazinol B(10),stigmast-4-en-3-one(11),catechin(12),gallic acid(13),luteolin-7-O-β-D-glucoside(14).HPLC method for determination of oleanolic acid in C.huchouensis,HPLC chromatography conditions: the mobile phase was acetonitrile-water(90:10),detection wavelength 210 nm,flow rate 1.0 m L / min,column Temperature is 25 °C,and the content of oleanolic acid in the self-made 9 extracts of C.huchouensis extract was calculated to be between 0.46% and 0.67%.The best water extraction process for C.huchouensis is to add 12 times water,reflux for 1 h,extract 3 times,combine the extracts,filter,concentrate the filtrate to thick paste,and dry to remove dry extract.According to the prescription amount,the original medicinal material is extracted by the optimal water extraction process,and the extract is obtained.And the appropriate matrix and auxiliary materials are added thereto to prepare the C.huchouensis cream,which has uniform texture,fine appearance,easy cleaning and stable quality.Conclusion:14 monomer compounds were isolated and identified in C.huchouensis.Compounds 1 to 14 were isolated from C.huchouensis for the first time.Compound14 was isolated from the genus Clematis for the first time.The method for determining the oleanolic acid content in C.huchouensis is precise,stable,feasible and reproducible.The extraction process of C.huchouensis is stable and reliable,reasonable and feasible.The preparation process of C.huchouensis cream is simple,stable and feasible.