粗多糖提取及纯化方法.docVIP

粗多糖提取方法 （1）玉米须原料用80%乙醇在78提取三次，去除色素、单糖和脂溶成分。过滤，合并滤渣，使乙醇挥发干净，粉碎滤渣。 用100热水浸提干燥玉米须一小时，料液比1：15，提取三次。合并提取液、过滤和浓缩，利用80%乙醇醇沉。 离心收集醇沉物 (3000 ×g，10min)，然后用水溶解，并利用Sevag方法去除游离蛋白质。 脱去蛋白质的溶液再次用80%乙醇醇沉。收集沉淀，先后用无水乙醇和丙酮溶液冲洗三次。最后冷冻干燥，获得玉米须粗多糖。Hot?water?extraction?of?crude?polysaccharides All?the?corn?silk?raw?materials?were?immersed?in?80%?(v/v) ethanol at?78℃?for?three?times?to?remove?colored?materials, monosaccharide and?liposoluble?constituents.?The?organic?solvent was?volatilized?and the?pretreated?corn?silk?was?obtained?to?crush. Dried?ground?corn?silk?(100?g)?was?extracted?with?water?at?100℃ (1:15?(w/v),?1?h,?3?times).?The?extraction?solutions?were?combined, filtered?and?concentrated,?precipitated?by?the?addition?of?anhydrous?ethanol?to?a?final?concentration?of?80%? (v/v). Precipitates were?collected?by?centrifugation?(3000?×?g,?10?min),? then?dissolved with?water,?and?subjected?to?the?Savage?method?(chloroform: butyl alcohol,?4:1)?to?remove?free?proteins.?The?deproteinized?solution was?re-precipitated?in?80%?(v/v)?ethanol.?The?precipitates?were collected?and?successively?washed?with?anhydrous?ethanol?and acetone.?After? freezing?dried,?the?hot?water?extraction?corn?silk polysaccharides?(PS)?were?obtained. 第二种方法 Preparation of ILPSThe preparation of crude ILPS was carried out according to thereported method with some modifications (Xu et al., 2012). Briefly,the powder of I. latifolia Thunb was pre-extracted two times with85% aqueous ethanol solution (v/v) in a ratio of 1:15 (materialto ethanol solution, g/mL) at 80?C for 1 h each, and the super-natant (containing colored materials, oligosaccharides and smallmolecule compounds) was removed. The resulting residues wereextracted three times with distilled water in a ratio of materialto water 1:20 (g/mL) at 90?C for 3 h each, and the extracts werecentrifuged at 5000 rpm for 15 min. The supernatants were com-bined and concentrated by a rotary evaporator to a proper volume.The resulting concentrate was mixed with three times volume