Study On Chemical Composition Of Propolis’ Hypoglycemic Effective Part

Propolis is Italian bees’(Apis mellifra L.) dry secretion, its appearance are round or irregular fragments, most are brown, dark brown or beige, with luster, fragrance, tasting bitter and hot feeling. It contains abundant of flavonoids, and the flavonoids have a variety of physiological roles:lower blood fat andsugar, improve microcirculation, improve body’simmunity, diminish inflammation, eliminate sputum, antipyretic, detumescence, antibacterial, antiviral and antifungal, surpress and destroy the tumor, etc.Research group’s early work showed that propolis’ ethanol extraction had an anti-diabetics activity. Applicated tracking method to determine the activity of lowering blood sugar effective part, found that the part’s total flavonoids was the part’s main ingredients.This thesis chosen propolis bought from Henan province as the research object, used orthogonal experimental design and the method of synthetical weighted score of data analysis to optimizate total flavonoids in propolis’best extract technology which could improve the content of flavonoids, then we got propolis’hpyerglycemic effective part. In order to furthering clarify the chemical basis of propolis’ hpyerglycemic effective part, we integrated use silica gel column chromatography, preparation liquid chromatography and Sephadex LH-20column chromatography methods, finally nine single compounds were separated from propolis’s effective part. In order to determine all nine single compounds, we comprehensive used UV, EI-MS,13CNMR and’HNMR datas, respectively were:izapinin(Ⅰ), tectochrysin(Ⅱ), pinocembin(Ⅲ), galangin(Ⅳ), chrysin(Ⅴ), kaempferol(Ⅵ), quercetin(Ⅶ), ferulicacid(Ⅷ),3,5,7-trihydroxyl-4’-methoxyl flavonol (Ⅸ).For studying pharmacological activity of propolis’hpyerglycemic effective part further, we made use of streptozocin and alloxan to cause mice and rats high blood glucose models to set up pharmacological experiments. Determination the levels of normal mice’s blood sugar and high blood sugar mice model’s glycogen, total cholesterol, insulin and other indexes. Finally, the pharmacological experiments showed that propolis’hpyerglycemic effective part had a good effect on lowering blood glucose and it could be produced into drugs.Get the following conclusions:1. The Henan propolis’best extraction condition was:three-time exaction for townty-four hours each time gave an optimal extraction of total flavonoids from proplis at a material/liquid ratio of1:10with90%ethanol as extraction solvent. Then, we used thirty-five times amount of1%sodium hydroxide to dissolve propolis ethanol-extraction, added strong hydrochloric acid to adjust the alkali solution’s pH to seven, it produced a large amount of precipitation. Over a night, we filtered the solution and precitation, dried the precipitation with pressure, rubbed it into small powder, and determined the content of total flavonoids was more than50 percent.2. Used different organic solvents to extract propolis’ fall blood sugar effective part. Then, we comprehensive used of silica gel column chromatography, sephadex LH-20column chromatography, half preparation liquid column chromatography and other methods to separate it, finally we got nine single compounds from the active part. At last, through EI-MS,13CNMR,1HNMR and HPLC analysis methods identified all nine single compounds.3. Used that streptozocin and alloxan caused mice and rats high blood glucose models, compared to the normal mice, these mice and rat models were given propolis flavonoids of20-160mg/kg different doses through orally, surveyed that normal and high blood glucose mice models’ blood glucose, liver glycogen, total cholesterol and insulin levels. The results showed that all dose groups of propolis total flavoniods had a good effect on falling blood sugar of streptozocind and alloxan models. So propolis’ total flavonoids had a good lowering blood sugar effect, it could provide references for the further study and development of propolis in the prevention and treatment of diabetes mellitus.This study based on a systemic, comprehensive base that studied on prepared the propolis’ effective parts, analysis the chemical composition of effective parts, established animal models to study propolis’fall blood sugar activity, all those could lay a experimental foundation for the development of independent intellectual property rights of the four categories of new drugs.