The Research On The Extraction, Isolation And Identification Technology Of Flavonoids From Chrysanthemum And The Activeantibacterial Property

Xinjiang snow daisy (Chrysanthemum) scientific name coreopsis (C. drummondii Torr. et Gray), alsoknown as "Alpine xiangju", belongs to Asteraceae (Compositae) and which is an annual dried flower heads(Ctinctoria) are relatives of herbs. The name of Uighur is "Guli"(Gulqai). As early as the Yuan Dynastywhen there is relevant reports of snow daisy used as an ingredient of Xinjiang Qijudihuang pills to controldiabetes. Snow Chrysanthemum is medicinal and edible plant, the study of Xinjiang snow daisy is of greatsignificance to medicinal resources in Xinjiang.The experiment takes flavonoids the Xinjiang snow daisy contained as the research object, usingultrasonic to extract total flavonoids and using response surface methodology to optimize flavonoids ofsnow daisy extraction process. The experiment use normal pressure silica gel column chromatography toisolate flavonoids, and take some tests to monomer and total flavonoids, for example of in vitro and in vivoantioxidant activity tests, the study of hypoglycemic activity and antibacterial activity etc. The purpose is toscreen the active ingredient (monomers) of snow chrysanthemum.(1) The test uses a full range of chemical pre-test test method to preliminary study the chemicalcomposition of snow daisy. The results showed that: snow daisy may contain the following substances:flavonoids contain isoflavones, flavanones, flavonols dihydrogen5-hydroxy flavones,2’-hydroxychalcone, catechol hydroxyl, alkaloids, anthraquinones contain hydroxyanthraquinone class, saponins,carbohydrates, phenols, coumarins, terpenes, proteins, organic acids and so on. Total flavonoid contents ofsnow daisy of flowers, stems and leaf measured is: inflorescence21.95%> stems16.29%> leaves10.53%,The results showed that the content of flavonoids in inflorescence parts becomes higher levels Therefore,the choice of materials for the subsequent trials is snow daisy sequence as the test material.(2) Optimizing the extraction process of total flavonoids in snow daisy: taking rutin as referencesubstance and using UV spectrophotometry to test the total flavonoids content of snow daisy, usingultrasonic extraction, optimizing extraction process to determine optimum extraction parameters by singlefactor response surface method. The optimum conditions were:80%ethanol, solid-liquid ratio1:20g/mL,extraction time30minutes, ultrasonic power50W. Under this condition, the extraction rate of totalflavonoids is24.9mg/mL.(3)The purification, isolation and structure identification of total flavonoids in snow daisy, purified bymacroporous resin AB-8and silica gel column chromatography to obtain a pure product-snow daisy XJ-1,and the first time to isolate from snow daisy. By TLC, HPLC-MS and NMR, snow daisy XJ-1is identifiedto be Okanin.(4) Vivo and vitro antioxidant test. The experiments with mice for the study, to evaluate snow daisyflavonoids Vivo and in vitro antioxidant capacity from the ability of clear the ABTS+, O2-nd OH ndDPPH radical capacity. Compare the snow daisy crude flavonoids and flavonoids or purified flavonoidsextracted with different solvents to evaluate antioxidant properties. In vitro results showed that:The testresults of Clearing O-2radical of Snow daisy flavonoids XJ-1: XJ-1>VC>ethyl acetate material> n-butanolphase material> chloroform phase material>Water Matter>crude flavonoids; ABTS test results: XJ-1>ethylacetate material>n-butanol phase material>VC>chloroform phase material>Water Matter> crudeflavonoids; OH est results: XJ-1>ethyl acetate material>VC>n-butanol phase material> chloroform phase material>Water Matter> crude flavonoids; DPPH Test results: XJ-1>ethyl acetate material>n-butanol phasematerial>VC>chloroform phase material>Water Matter> crude flavonoids; In summary, test results showthat in vitro antioxidant capacity XJ-1is stronger than VC. In vivo results showed that: XJ-1>snow daisyhigh levels of flavonoids group>VEGroup> snow daisy middle levels>snow daisy flavonoids low volumegroup.(5) Lowering blood sugar and antibacterial effects of flavonoids: the capacity of lowering bloodsugar of chrysanthemum flavonoids XJ-1is50mg/Kg.In addition, the experiment carried out total flavonoids and snow daisy flavonoids XJ-1bacteriostasison Staphylococcus epidermidis, results show that the snow daisy flavonoids XJ-1has a strong inhibitoryeffect to Staphylococcus epidermidis compared to total flavonoids.