荞麦麸皮多糖的分离纯化及其外貌表征的分析
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摘要
采用水提醇沉法对荞麦麸皮多糖(Buckwheat bran polysaccharide,BBP)进行提取,通过DEAE-52纤维素柱层析法和Sephadex G-200对其进行分离纯化,采用凝胶过滤法对多糖组分的分子量进行测定,采用扫描电镜对荞麦麸皮多糖及其分离纯化的多糖组分的外貌进行分析,用红外光谱对分离纯化的三个荞麦麸皮多糖组分的苷键构型进行分析。结果表明:DEAE-52纤维素和Sephadex G-200对荞麦麸皮粗多糖具有较好的分离纯化效果,得到了三个多糖组分(BBP-Ⅰ、BBP-Ⅱ和BBP-Ⅲ),其分子量分别为1.56×10~4Da、5.85×10~4Da、8.45×10~4Da。扫描电镜的结果表明,BBP其表面粗糙,凹凸不平,三个分离纯化的多糖组分形貌差异较大,分别以球状、片状、棒状为主。红外光谱扫描表明BBP-Ⅰ和BBP-Ⅲ为ɑ-糖苷键化合物,BBP-Ⅱ为β-糖苷键化合物。荞麦麸皮多糖组分外貌的差异性为其进一步的结构探究提供了重要依据。
In this work,Buckwheat Bran Polysaccharides( BBP) was extracted by water extraction and alcohol precipitation method. It was further purified by DEAE-52 cellulose column chromatography and Sephadex G-200. The molecular weight of the purified product was determined by gel filtration method,followed by the morphology characterization and component analysis using electron microscopy. The glycosidic bond configuration of the purified product was analyzed by infrared spectroscopy. The results showed that three polysaccharide fractions( BBP-I and BBP-II and BBP-III) were obtained by DEAE-52 cellulose column chromatography and Sephadex G-200 separation and purification and their molecular weights were 1. 56 × 10~4 Da,5. 85 × 10~4 Da,8. 45 × 10~4 Da,respectively. The electron microscopy showed that the surface of BBP was roughness and uneven. There was significant diversity about the appearance of the purified product,which mainly characterized by spherical,lamellar and rod,respectively. IR spectra demonstrated that BBP-I and BBP-III were alpha glycosidic bond compounds,while BBP-II was beta glycosidic bond compound. The differences of polysaccharide components in buckwheat bran would provide an important foundation for further structural research.
In this work,Buckwheat Bran Polysaccharides( BBP) was extracted by water extraction and alcohol precipitation method. It was further purified by DEAE-52 cellulose column chromatography and Sephadex G-200. The molecular weight of the purified product was determined by gel filtration method,followed by the morphology characterization and component analysis using electron microscopy. The glycosidic bond configuration of the purified product was analyzed by infrared spectroscopy. The results showed that three polysaccharide fractions( BBP-I and BBP-II and BBP-III) were obtained by DEAE-52 cellulose column chromatography and Sephadex G-200 separation and purification and their molecular weights were 1. 56 × 10~4 Da,5. 85 × 10~4 Da,8. 45 × 10~4 Da,respectively. The electron microscopy showed that the surface of BBP was roughness and uneven. There was significant diversity about the appearance of the purified product,which mainly characterized by spherical,lamellar and rod,respectively. IR spectra demonstrated that BBP-I and BBP-III were alpha glycosidic bond compounds,while BBP-II was beta glycosidic bond compound. The differences of polysaccharide components in buckwheat bran would provide an important foundation for further structural research.
引文
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2 Lin R,Zhou Y,Wang R,et al.A study on the extract of tartary buckwheat.Toxicological Safety of the Extract of Tartary Buckwheat.Adv Buc Res,2001:602-607.
3 Zang Z,Yang B,Shen Q,et al.Buckwheat production,research and market in China.Adv Buck Res,2001:690-693.
4 Xu LH(徐丽华),Pan H(潘宏),Zhao YM(赵英明).Buckwheat-a new kind of multipurpose crops.Rain Fed Crops(国外农学-杂粮作物),1998,18(3):1-4.
5 Wang Q,Ogura T,Wang L.Research and development of new products from bitter-buckwheat.Cur Adv Buck Res,1995,873-879.
6 Zhao G,Tang Y,Hu Z,et al.Selection and application of tartary buckwheat new variety Xiqiao One.Adv Buck Res,2001:382.
7 Zong A,Cao H,Wang F.Anticancer polysaccharides from natural resources:A review of recent research.Carbohydr Polym,2012,90:1395-1410.
8 Wasser S.Medicinal mushrooms as a source of antitumor and immunomodulating polysaccharides.Appl Micro Biot,2002,60:258-274.
9 Bi YP(毕研平),Su YF(苏艳芳),Chai X(柴欣),et al.The development of the chemical constituents and biological activity of buckwheat.Northwest Pharm(西北药学杂志),2008,32:116-118.
10 Lu JJ(路静静),Zhao YQ(赵余庆).Research progress on components,bioactivity and applications of buckwheat Hulls.Food Res Dev(食品研究与开发),2016,22:210-213.
11 Xi XG,Wei XL,Wang YF,et al.Determination of tea polysaccharides in Camellia sinensis by a modified phenol-sulfuric acid method.Arch Biol Sci,2010,62:669.
12 Fan SH(范三红),Liu YR(刘艳荣),Yang ML(杨民乐),et al.Separation,purification and structural analysis of polysaccharide from Chinese pine pollen.Food Sci(食品科学),2009,30(15):127-129.
13 Xu WT(许文涛),Zhang FF(张方方),Luo YB(罗云波),et al.Isolation and purification of water-soluble polysaccharide from Common Buckwheat(Fagopyum esculentum)and determination of its molecular weight.Food Sci(食品科学),2009,30(13):22-24.
14 Yang YJ(杨勇杰),Jiang RZ(姜瑞芝),Chen YB(陈英红),et al.Determination of sugars in heteropolysaccharide by phenol-sulfuric acid method.Chin Tradit Pat Med(中成药),2005,6:706-708.
15 Lee JS,Kwon JS,Yun JS,et al.Structural characterization of immunostimulating polysaccharide from cultured mycelia of Cordyceps militaris.Carbohydr Polym,2010,80:1011-1017.
16 Su H(苏浩).Study on extraction,structure identification and physical properties of water-soluble soybean polysaccharide from soybean dregs.Xinjiang:Xinjiang Agricultural University(新疆农业大学),MSc.2009.
17 Liang T(梁涛),Zhang J(张静),Zhang LN(张力妮),et al.Preliminary structure analysis and structure morphology observation of alkali-extractable polysaccharide PEAP-1 from Pleurotus eryngii.J Food Sci Biotech(食品与生物技术学报),2013,32:951-956.
18 Su Y(苏瑛),Zheng XC(郑晓翠),Liu PJ(刘鹏举),et al.Purification and determination of the structure of Epimedium polysaccharide.Food Ferment Ind(食品与发酵工业),2009,35(12):93-96.