Extraction Separationand Identification Theactive Ingredients Of Brassica Napus L. Pollen

Brassica Campestris L. is called chinese cabbage ,which belongs to the cruciferous plant and is the leaves of tender bine rape seed. People plants it throughout the north and south of China as oil crop, and always has a good harvest. Brassica napus L. pollen is the bee collect from the stamen of Brassica Campestris L., and joined the special gland secretion,nectar and Bees saliva.Bee pollen is excellent natural nutritious food, is also a kind of ideal tonic, and prostate disease have very good medical function.This paper used bee pollen rape as research object.This project explored the chemical constitution and structural characteristics of pure flavonoid ,unsaturated fatty acids derivatives and alkaloid components of Brassica napus L. pollen.Laying the foundation for the development of Brassica napus L. pollen research.(1)The methods of extraction flavonoid from Brassica napus L. pollen.Selected the ratio of solid to liquid, ethanol concentration ,extracting time and extracting temperature four factors as the solvents affecting of total flavonoid.Through the single factor and orthogonal methodology was used to optimize the extraction of total flavonoid from Brassica napus L. pollen. The best extraction technology of total flavonoid was:ethanol concentration 95%, temperature 60℃, the ratio of solid to liquid(M/V)l:15, extracting time 2.0 hour ,and the yield of flavonoids was up to 3.27% .(2)The isolation and identification of flavonoid compounds.Broken bee pollen through petroleum ether,ethyl acetate,butyl alcohol.Then we get the crude extract of bee pollen.The bee pollen butyl alcohol crude were separated and purified by polyamide column chromatography ,ODS reversed-phase column. Appraised by thin-layer chromatography andThe structures were identified by NMR,MS. Finally, three major flavonoid compounds 1, 2 and 3 were extracted. The compound 1 is Kaempferol-3-O-β- D pyranoid glucose base (1 ,2)-β-D-pyranoid glycosidase,the compound 2 is Quercetin-3-O-β-D-pyranoid glycosidase,the compound 3 is Kaempferol-3-O-β-D-pyranoid glycosidase. (3)The isolation and identification of unsaturated fatty acid derivatives. Using MCI and ODS chromatographic column separated and purified the ethyl acetate extract of Brassica napus L. Pollen.Finally,we got three long chain aliphatic acids derivatives.The use of NMR method to identify their structures. Compound 4 is 9,12,15-octadecatrienoic acid, Compound 5 is(2-(2,3-dihydroxypropoxy)ethyl)carbamic(9E,12E,15E)-octadeca-9,12,15-trienoicanhydrid e, Compound 6 is (2-(3-hydroxy-2-methoxypropoxy)ethyl)carbamic(9E,12E,15E)-octadeca-9,12,15-trienoic anhydride.Compound 5 and 6 are the first time isolate from bee pollen rape.(4) The isolation and preliminary identification of Alkaloid.The use of column chromatography separated the ethyl acetate extract. We got two single compounds 7 and 8. Identified by NMR,and confirmed is a nitrogen compound,which structure need further identification.