Study On The Antioxidant Activity Compositions Of Rape Bee Pollen

Rape bee pollen is the pollen load collected by bees from rape and mixed with nectar and bees’ secretion during the collecting process. It has the efficiency of anti-oxidation, enhance immunity and prevention and treatment of prostate disease and so on, widely used in the field of food, cosmetics, health products and clinical medicine and so forth. Currently, the research on material basis of antioxidant effect of bee pollen is not perfect, therefore in this paper we carried out studies as follows to study the antioxidant activity compositions of rape bee pollen more comprehensively.(1) The antioxidant and immune activity of different components of rape bee pollen were studied. Rape bee pollen was extracted by 70% ethanol after powdered, acquired the ethanol extract. The filter was extracted by deionized water, acquired the water extract. The ethanol extract was successively extracted by petroleum ether, ethyl acetate and butanol, obtained the rape bee pollen petroleum ether extract, ethyl acetate extract and butanol extract. The activity of all these components of rape bee pollen were tested through the D-galactose induced aging model mice test, results showed that the petroleum ether extract and the ethyl acetate extract both have significant antioxidant activity, but the butanol extract and water extract don’t have significant antioxidant activity neither. The water extract has significant immune activity, but the other tree extracts haven’t significant immune activity all.Ethanol sediment was prepared from water extract, and it has significant immune activity which was confirmed through the immune suppression mice test. Then ethanol sediment was separated by the macroporous resin chromatography column, DEAE-cellulose column,Sephacryl chromatographic column and other chromatographic column, acquired one main polysaccharide compound, its structure was analysed by the NMR technology, obtained that its structural formula is-[HOCH2-(CHOH)4-COOH]-, and this polysaccharide is an rare pollen polysaccharide alcohol compound. The immune suppression mice test showed that this polysaccharide compound has significant immune activity.(2) The separation and structure identification of petroleum ether extract. The petroleum ether extract of rape bee pollen was separated by MCI chromatography column and ODS chromatography column, acquired two main compounds 1 and 2. Analysing the structure of compound 1 and compound 2 by NMR technology, results showed that compound 1 isα-linolenic acid, and compound 2 is α-linolenic acid monoglyceride.(3) The constitutes and antioxidant activity of ethyl acetate extract were studied. The acetate extract of rape bee pollen was separated into 30% and 50% ethanol elution part by MCI chromatography column, and the D-galactose induced aging model mice test showed that both the two parts have significant antioxidant activity. Compound 3 and 4 were separated from 30% ethanol elution part, then using the NMR technology to analyse theirstructure, learned that compound 3 is quercetin-3-O-β-D-glucopyranoside, and compound 4 is kaempferol-3-O-β-D-glucopyranoside. 50% ethanol was roughly separated into component A and component B, and compound 5 and compound 6 were purified from these two components, using the NMR technology to analyse the structure of these two compounds,result showed that these two compounds were not single composition. Therefore component A and component B were re-separated in the next chapter.(4) The separation and structure identification of component A and component B. The D-galactose induced aging model mice test showed that both the two components have significant antioxidant activity. Their compositions were analysed by the UPLC technology,then using the MCI chromatography column and ODS chromatography column to separate them, both acquired two compounds from component A and component B, namely compound A2, A4, B1 and B2. Analysing the structure of these four compounds by the NMR technology,results showed that the structure formula of compound A2 is HO-C6H4-CH=CH-CO-NH-(CH2)4-NH2, and the other tree compounds’ structure are familiar with compound A2, but their specific structure need further analysis.