【作者】 陈传福；

【导师】 张培正；

【作者基本信息】 山东农业大学 ， 食品科学， 2008， 硕士

【摘要】 熟地黄多糖是“药食同源”熟地黄中特有的活性成分之一,药理研究表明,熟地黄多糖有增强免疫、抗肿瘤、补血等生理功能。研究熟地黄多糖的提取、纯化工艺及其结构性质,对熟地黄资源的深加工和熟地黄多糖食品、保健品的开发有重要意义。本文在研究加热法提取熟地黄多糖的基础上,将超声技术引入熟地黄多糖的提取,用响应面设计法寻找超声提取的最佳工艺;并利用SephadexG-200葡聚糖凝胶与DEAE-Sepharose柱层析纯化熟地黄多糖,在结构分析中,采用了紫外光谱,红外光谱和薄层色谱。熟地黄多糖加热提取实验表明,加热提取熟地黄多糖的最佳条件:颗粒粒度40目、浸体温度90℃、料液比1:70、浸提时间3h、此时熟地黄多糖的提取率为6.679%。以颗粒粒度40目熟地黄为原料超声提取熟地黄多糖能显著提高生产效率并简化提取工艺,其最佳条件为:料液比1:56,提取温度64.5℃,提取时间3.1min,熟地黄多糖的提取率为6.801%。得出回归方程:y=6.6463+0.304x1+0.1415x2+0.212x3-0.061x1x2-0.2872x1x3-0.0815x2x3-0.2527x12+0.0665x22-0.2727x32熟地黄多糖分离纯化实验表明:熟地黄多糖提取液浓缩后用sevage法除蛋白,再用蒸馏水透析,经醇沉后的熟地黄多糖,冻干后备用。SephadexG-200葡聚糖凝胶纯化的的熟地黄多糖只有1个洗脱蜂,经DEAE-Sepharose纯化的熟地黄多糖呈现2个洗脱蜂,说明DEAE-Sepharose比SephadexG-200葡聚糖凝胶具有高的分辨率。在纯化过程中SephadexG-200葡聚糖凝胶几乎对色素无吸附作用,而DEAE-Sepharose对色素具有很强的吸附作用,因此,本实验选择DEAE-Sepharose纯化熟地黄多糖。熟地黄多糖的单糖分析:采用薄层色谱分析法对熟地多糖的单糖组成进行了分析。将熟地黄多糖完全酸解后,采用硅胶薄层板,以正丁醇:乙酸:水(9:4:2)为展开剂,10%硫酸-乙醇溶液为显色剂。分析结果表明:熟地黄多糖主要由葡萄糖、半乳糖、果糖和水苏糖等单糖组成。更多还原

【Abstract】 Rehmannia glutinosa polyses(RGP) exists particularly in Radix Rehmannia Preparate,which not only is food ,but is chinese tradtional medicine.Pharmacological investigations indicated RGP can resist tumor、adjust immunity,increase blood and so on.To study the extracting technology of and purification and structure are important to deeper the process of Radix Rehmannia Preparate and the development of functional food and medicine of RGP.This paper based on studying hot extracting of RGP, the supersonic was applied to strengthen the extraction and the supersonic extracting was optimized using response surface methodology. The crude polysaccharides were isolated and purified by chromatographying with SephadexG-200 and DEAE-Sepharose column. During the structure analysis of RGP, purity and component sugars were carried out by UV, IR, TLC.The extracting test showed that the best hot extracting technology of RGP was that:the particle of 40 mesh size;extraction temperature90℃sample ratio : solvent=1:70(m/V);extraction time 2h , the extracting rate is 6.679%.Supersonic extracting can improve the extracting rate and reduce the extracting time and simplify the extracting process. The better supersonic extracting technology was that: the particle of dried Radix Rehmannia Preparate 40 mesh size;sample ratio : solvent=1:56(m/V); extraction time 3.1min and extraction temperature 64.5℃. The extracting rate is 6.801%. After regression analysis , factors which influenced response variables significantly new selected. Consequently multinomial regression equation was obtained: y=6.6463+0.304x1+0.1415x2+0.212x3-0.061x1x2-0.2872x1x3-0.0815x2x3-0.2527x12+0.0665x22-0.2727x32The separation and purification technology of RGP test showed: The extracting solution of RGP concentrated by vacuum rotary evaporator, then eliminated protein using sevage reagent and dialysed by distilled water. When ethanol of certain concentration added, many white deposit were formed. Then the deposit dried by vacuum freeze drier. The crude polysaccharides were isolated and purified by chromatographying with SephadexG-200 and DEAE-Sepharose column. DEAE-Sepharose column had a higher partition rate. During the purification process, DEAE-Sepharose column had more adsorption for pigment than SephadexG-200 column. Therefore, DEAE-Sepharose column was chosen for the purification of RGP.Composition analysis of Rehmannia glutinose Polysaccharides was studied. The TLC method was established to analysis composition of Rehmannia glutinose Polysaccharides Silica gel plate, n-butanol : HAc : H2O (9:4:2) as developing solvent, 10%sulfuric acid-ethanol eoloration. Rehmannia glutinose Polysaccharides was composed of galactose, glucose, fructose and stachyose.更多还原