Study On The Extraction And Separation And Determination Of Chemical Constituents From Folium Artemisiae Argyi

Objectives:To extract of flavonoids from Folium Artemisiae Argyi, and establish a assay method for determination the total flavonoid content of Folium Artemisiae Argyi. To extract and isolated of flavonoid and other chemical constituents from Folium Artemisiae Argyi, optain pure componds as well as identify the chemical structure, offer the accordance for Systemic research on the efficacy, toxicity and chemical composition of Folium Artemisiae Argyi. Establishment of this method laid a good foundation for a large number of extractions and preparations of monomeric compounds in the Folium Artemisiae Argyi as well as further developmentMethods:In this study, on the basis of consulting lots of relevant literature, the structural characteristics and chemical properties of the chemical composition of Folium Artemisiae Argyi were analysis anthetically,then development of experimental. Firstly, by means of ethanol immersion to extract the total flavonoids, concentrate them into lipstick. The color reactions of the solution which is made of Pre-test materials from the extracts with Sodium nitrite and aluminum nitrate were determined the absorbance values by UV. This paper analyzed influencing factors, such as adding time of color development reagent, pH of solution, selected the best experimental approach, drew the standard curve and study on the methodology so as to establish the determination method for the total flavonoid content of Folium Artemisiae Argyi.Then,extract the lipstick by solvents with different polarity (Petroleum ether, Ethyl acetate, Butanol). Then by repeated column chromatography, including Silica gel,Sephadex LH-20, Toyopearl HW-40, PTLC and HPLC,4 compounds were isolated from the EtOAc extract of Folium Artemisiae Argyi. The isolated compounds were identified by means of 1H-NMR,13C-NMR and 2D-NMR spectra.Results:By analyzed of influencing factors and studied on the methodology, the determination method of total flavonoid content of Folium Artemisiae Argyi was established:standard curve:A=0.0118C+0.0133, r=0.9991,good linear relationship in the range of 5.800～58.000μg/ml. The concentration of total flavonoid:39.06μg/ml. This determination method shows good quality of accuracy, precision, and repeatability and good stability within 30 minutes. There were 4 compounds,5,7, 3'-trihydroxy-3,6,4'-trimethoxyflavonol(1),5,7-dihydroxy-3,3',6,4'-tetrame-thoxyflavonol(2), scopoletin(3), p-hydroxylben-zaldehyde(4), were isolated from the EtOAc extract of Folium Artemisiae Argyi. Compound (1) and (2) belongs to flavonoids,compound (3) belongs to Coumarin. And compound (4) belongs to Aromatic Mixture.By literature retrieval, compound (1) and (2) were frist isolated from Folium Artemisiae Argyi.Conclusions:Based on the color theory of sodium nitrite and aluminium nitrate, the UV-spectrophotometric method is simple, quick, accurate and stable in measuring absorbance, which can be used as the assay method for determination the total flavonoid content of Folium Artemisiae Argyi.In this study, chemical compositions of Folium Artemisiae Argyi were separated and purified using chemical extraction and separation method for a variety of naturally occurring drugs, and the compound was separated and then performed structural identification by modern spectroscopy technique. Finally, four kinds of monomeric compounds were determined, which supplied preliminary study for chemical compositions of Folium Artemisiae Argyi. This method had maneuverability and was more accurate and effective compared with traditional extracting method with alcohol-water system.Compounds (1) and (2), which have been separated and purified were identified as flavonoid derivatives, additionally they were firstly separated from this plant, which not only made up lack of understanding flavanoid during modern study of Folium Artemisiae Argyi to some extent, but also provided the necessary prerequisites for pharmacological activity scanning as well as toxicity study. With the development of research, the knowledge of chemical compositions and pharmacological activity of Folium Artemisiae Argyi will be clearer. In order to elucidate the relationship of drug action, it was found that lead compound with drug activity provided basic data, which inevitably promoted the research of the modernization of traditional Chinese medicine, Folium Artemisiae Argyi, as well as its further development and utilization.