异枝麒麟菜（Eucheuma striatum）硫酸多糖的超声辅助提取工艺及抗氧化活性研究
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摘要
本文包括两部分,一部分是以南海大型经济海藻异枝麒麟菜(Eucheuma striatum)为原料,对其生物活性硫酸多糖的超声辅助提取工艺进行了优化,并对其分离纯化、理化性质进行分析测定,对多糖样品的体外抗氧化活性进行了初步的研究和探讨,为海藻资源的综合开发利用提供科研理论依据。另一部分是对瑶药翠云草石油醚萃取部位化学成分进行分离。
对异枝麒麟菜硫酸酯多糖(ESSP)的超声辅助提取工艺进行研究,通过单因素实验和正交试验对该提取工艺进行优化。实验表明,超声时间是提取硫酸酯多糖的关键影响因素。最佳工艺条件为超声时间20min,超声功率150W,料液(水)比1：90(g／mL),多糖的提取率达48.14%,所得的多糖含量96.94%,硫酸酯基含量19.66%。并与热水提取法进行了比较,采用优化工艺提取的多糖提取率、多糖含量和硫酸酯基含量比热水提取法分别提高了12.08%、8.07%和1.41%。
采用DEAE-52纤维素离子交换柱对粗多糖ESSP进行分离纯化,然后经交联琼脂糖凝胶Sephadrose CL-6B柱进一步纯化得到ESSP I、ESSP II,凝胶渗透色谱测定其相对分子量分别为：ESSP I的重均分子量Mw为2.0114×106Da; ESSPⅡ的重均分子量为Mw为1.185×106Da。红外谱图显示ESSP I和ESSP II均有多糖类物质的特征吸收峰。
采用Fenton反应和邻苯三酚自氧化体系探讨了异枝麒麟菜多糖的体外抗氧化效果。结果发现异枝麒麟菜多糖具有较好的清除氧自由基O2·-的活性,尤其是其清除·OH的作用更加显著。实验发现异枝麒麟菜多糖纯化后的组分ESSPⅠ的抗氧化活性要比粗多糖高,而且多糖的作用活性与其浓度有剂量-效应关系。
同时还对瑶药翠云草的石油醚萃取部位进行了分离,得到4个单体化合物,采用光谱分析和文献对照的方法对分离所得单体的结构进行鉴定,分别为：大黄素甲醚(1),红灰青素(2),木栓酮(3),β-谷甾醇(4)。以上化合物均为首次从该植物中分离得到。
The thesis summarizes including two parts. One part, a comprehensive procedure for ultrasonic-assisted extracting active polysaccharides from Eucheuma striatum, a kind of big economical algae in South-China Sea, had been developed. The separation, physical and chemical properties of sulfated polysaccharides had been determined. The antioxidant activities of the polysaccharides was discussed. This conclusion could provide theoretical basis to the comprehensive development and utilization of marine algae resources. Another part is about the separating from the Petroleum ether fraction of Selaginella uncinata (Desv.) Spring.
The ultrasonic-assisted extracting process of Sulfated polysaccharides from Eucheuma striatum was studied. One-factor experiment and orthogonal experiment were investigated. Extracting time had notable influence on the yield of polysaccarides. The optimum technology conditions were as follows: extracting with ultrasonic for 20min, ultrasonic power of 150w, dry material and water in a ratio of 1:90g/ml. With this optimum condition, the extraction ratio of polysaccharide is up to 48.14%, the polysaccharide content is 96.94% and the sulfate group content is 19.66%. Compared with the hot water extraction method, the extraction ratio of the polysaccharide, polysaccharide content and sulfate content extracted by the optimum technology are increased by 12.08%,8.07% and 1.41% respectively.
The ESSP were separated by DEAE cellulose-52 column chromatography, then the polysaccharides had a further purification by Sephadrose CL-6B column chromatography. The pure products were named ESSP I and ESSP II. GPC results showed that molecular weight of ESSP I was 2.0114×106Da and ESSP II was 1.185x106Da. IR spectrograms showed that ESSP I and ESSP II had characteristic absorption peaks of polysaccharides.
The anti-oxidation activities of the polysaccharides were studied by Fenton reaction and the self-oxidation of 1,2,3-phentriol. The results showed that the polysaccharides had better effects of elimination of O2·-, especially elimination of·OH. It was found that ESSP I was higher antioxidant activities than ESSP, and the role of polysaccharide concentration of activity with a dose-response relationship.
The chemical constituents of Petroleum ether fraction of Selaginella uncinata (Desv.) Spring was isolated. Their structures were elucidated by spectroscopic analysis and comparison with published data. Four compounds were obtained and characterized to be physcion(1), erythroglaucin(2), friedelin(3),P-sitosterol(4). All of the compounds were isolated from this plant for the first time.
对异枝麒麟菜硫酸酯多糖(ESSP)的超声辅助提取工艺进行研究,通过单因素实验和正交试验对该提取工艺进行优化。实验表明,超声时间是提取硫酸酯多糖的关键影响因素。最佳工艺条件为超声时间20min,超声功率150W,料液(水)比1：90(g／mL),多糖的提取率达48.14%,所得的多糖含量96.94%,硫酸酯基含量19.66%。并与热水提取法进行了比较,采用优化工艺提取的多糖提取率、多糖含量和硫酸酯基含量比热水提取法分别提高了12.08%、8.07%和1.41%。
采用DEAE-52纤维素离子交换柱对粗多糖ESSP进行分离纯化,然后经交联琼脂糖凝胶Sephadrose CL-6B柱进一步纯化得到ESSP I、ESSP II,凝胶渗透色谱测定其相对分子量分别为：ESSP I的重均分子量Mw为2.0114×106Da; ESSPⅡ的重均分子量为Mw为1.185×106Da。红外谱图显示ESSP I和ESSP II均有多糖类物质的特征吸收峰。
采用Fenton反应和邻苯三酚自氧化体系探讨了异枝麒麟菜多糖的体外抗氧化效果。结果发现异枝麒麟菜多糖具有较好的清除氧自由基O2·-的活性,尤其是其清除·OH的作用更加显著。实验发现异枝麒麟菜多糖纯化后的组分ESSPⅠ的抗氧化活性要比粗多糖高,而且多糖的作用活性与其浓度有剂量-效应关系。
同时还对瑶药翠云草的石油醚萃取部位进行了分离,得到4个单体化合物,采用光谱分析和文献对照的方法对分离所得单体的结构进行鉴定,分别为：大黄素甲醚(1),红灰青素(2),木栓酮(3),β-谷甾醇(4)。以上化合物均为首次从该植物中分离得到。
The thesis summarizes including two parts. One part, a comprehensive procedure for ultrasonic-assisted extracting active polysaccharides from Eucheuma striatum, a kind of big economical algae in South-China Sea, had been developed. The separation, physical and chemical properties of sulfated polysaccharides had been determined. The antioxidant activities of the polysaccharides was discussed. This conclusion could provide theoretical basis to the comprehensive development and utilization of marine algae resources. Another part is about the separating from the Petroleum ether fraction of Selaginella uncinata (Desv.) Spring.
The ultrasonic-assisted extracting process of Sulfated polysaccharides from Eucheuma striatum was studied. One-factor experiment and orthogonal experiment were investigated. Extracting time had notable influence on the yield of polysaccarides. The optimum technology conditions were as follows: extracting with ultrasonic for 20min, ultrasonic power of 150w, dry material and water in a ratio of 1:90g/ml. With this optimum condition, the extraction ratio of polysaccharide is up to 48.14%, the polysaccharide content is 96.94% and the sulfate group content is 19.66%. Compared with the hot water extraction method, the extraction ratio of the polysaccharide, polysaccharide content and sulfate content extracted by the optimum technology are increased by 12.08%,8.07% and 1.41% respectively.
The ESSP were separated by DEAE cellulose-52 column chromatography, then the polysaccharides had a further purification by Sephadrose CL-6B column chromatography. The pure products were named ESSP I and ESSP II. GPC results showed that molecular weight of ESSP I was 2.0114×106Da and ESSP II was 1.185x106Da. IR spectrograms showed that ESSP I and ESSP II had characteristic absorption peaks of polysaccharides.
The anti-oxidation activities of the polysaccharides were studied by Fenton reaction and the self-oxidation of 1,2,3-phentriol. The results showed that the polysaccharides had better effects of elimination of O2·-, especially elimination of·OH. It was found that ESSP I was higher antioxidant activities than ESSP, and the role of polysaccharide concentration of activity with a dose-response relationship.
The chemical constituents of Petroleum ether fraction of Selaginella uncinata (Desv.) Spring was isolated. Their structures were elucidated by spectroscopic analysis and comparison with published data. Four compounds were obtained and characterized to be physcion(1), erythroglaucin(2), friedelin(3),P-sitosterol(4). All of the compounds were isolated from this plant for the first time.
引文
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