Study On The Extraction And Purification Of Total Flavonoids From Fructus Canarii And Pharmacodynamics

Objective:The fresh Fructus Canarii as the research object, on the basis of the former study, First, screening the active ingredient in total flavonoids of Fructus Canarii extraction and purification methods and conditions, and determination; second, according to the pharmacological action of Fructus Canarii, mice as the research object, to study the total flavone for acute toxicity, anti-inflammatory and analgesia experimental; and study on in vitro antioxidant;Material foundation and scientific basis for research aimed at further separation of the total flavonoids and the preparation of monomer Methods:On the basis of the best Preliminary study, firstly using the best extraction method extracted total flavonoids from Fructus Canarii; study on purification of its crude extracts, screening the best separation conditions; based on the physical and chemical identification and the UV spectrum detection of the purified complexes, with physical and chemical characteristics and spectral characteristics of flavonoids, to be identified on purification may be the flavonoids. Followed the content of total flavonoids in the crude extract and purification was determine by UV spectrophotometry.Finally, to study the total flavonoids toxicology. Total flavonoids from Fructus Canarii intragastrically administered mice in experimental medicine group, with the largest volume permitted and the thickest concentration. The equal solvent administered mice in control group. The analgesic effect of total flavonoids from Fructus Canarii was observed by using the writhing assay and the hot-plate method in mice. To observe anti-inflammatory effects of total flavonoids from Fructus Canarii. Using three conventional methods includs egg white induced paw edema swelling, dimethylbenzene induced ear swelling and acetic acid induced celiac capillary permeability addition in mice were applied to evaluate the anti-inflammatory effects total flavonoids from Fructus Canarii. Detect the ability of eliminating or restoring total Flavonoids in Fructus canarii to Hydroxyl Radical and superoxide anion radical by three commom methods,1,10-phenanthroline-iron (Ⅱ) oxidation,1,2,3-Trihydroxy-benzene auto-oxidation and Prussia Blue spectrophotom. Results:We find the optimal purification conditions of total flavonoids in Fructus Canarii which was the solid-liquid ratio of1:10and70%ethanol reflux three hours at70℃water bath, the residues were extracted in duplicate3times, combined filtrate,filtered and solvent recovery.The optimum condition was the extract washed by water in Polyamide column,washed water was colorless,then washed by elution solvent was10%,30%,70%ethanol concentration,the elution velocity was3BV/h(bed volume,its abbreviation is BV),then the eluent which got from above conditions was dehydrogenized by vacuum reduced pressure drying method.the total flavonoids from Fructus Canarii had no acute toxicity was observed in both groups,The maximal tolerant dose (MTD) of total flavonoids from Fructus Canarii by ig in mice was11.46g·kg-1,which was about115times of adult clinical daily dose.Total flavonoids from Fructus Canarii reduced the times of writhing induced by intraperitoneal injection of acetic acid in mice, improved pain threshold in hot-plate.It could inhibit the mouse ear swelling induced by dimethylbenzene,inhibit the mouse paw edema swelling induced by egg serum,inhibit the increase of the mouse celiac capillary permeability induced by acetic acid.And it had the capacity for scavenging hydroxyl radical, superoxide anion and certain reducing power. Conclusion:screening the conditions of extraction and purification of total flavonoids from Fructus Canarii,we can get more purified flavonoids.Total flavonoids from Fructus Canarii has little acute toxicity,has good analgesic effect,has anti-inflammatory effect,has antioxidant activity in mice.The conclusion could provide a basis for further research of Fructus Canarii.