板栗多糖的提取、分离纯化及抗氧化活性研究

论文编号:SW095  论文字数:12629,页数:28

摘要：板栗营养价值丰富，具有养胃健脾，壮腰补肾的药用价值，国内对板栗多糖的研究鲜少报道。本文对去壳板栗经乙醇脱脂，热水浸提，醇析得到粗糖蛋白，并对提取得到的板栗粗糖蛋白进行脱蛋白工艺处理，得到板栗粗多糖4.42g，得率为1.77%。同时对板栗糖蛋白和板栗粗多糖进行了理化性质的测定，粗多糖在280nm处无吸收峰，说明蛋白质已基本除尽。考马斯亮蓝G-250测得板栗粗糖蛋白中蛋白质含量为2.28%，板栗粗多糖中蛋白质含量为0.026%。苯酚-硫酸法测得板栗粗多糖中己糖含量为89.54%。红外光谱显示，该粗多糖可能含有β-D-葡萄吡喃糖基。
     板栗粗多糖经Sephacryl S-100HR柱、DEAE-Sepadax Fast Flow阴离子交换柱层析分离后得到了8个板栗多糖组分。对板栗多糖组分CNPS-8进行纯度检验，采用层析仪经Sephacryl S-100HR洗脱，得到了一个单一对称的峰，证明该组分为一均一的精多糖。
     采用甲基紫模型，对板栗粗多糖进行羟基自由基清除率试验，结果证明板栗粗多糖在剂量为1-20mg/mL范围内,对羟基自由基具有一定的清除效果。且随浓度的增大，清除效果增强。但在同等剂量下弱于阳性对照Vc，20 mg/mL的板栗粗多糖的清除率为34.8%，而阳性对照Vc(20mg/mL)的清除率为66.08%。
关键词：板栗；多糖；提取；分离纯化；抗氧化活性  
Polysaccharides from Chestnut: Extraction, isolation,
 purification and antioxidant activity

Abstract: Being rich of nutrition, Chestnut has a lot of biological activity and high medicinal function, and it’s benefit to stomach, spleen, spinal and kidney. Domestic research of Chestnut polysaccharide was rarely reported. In this paper, crude polysaccharide is obtained by following process to peeled chestnuts: degreased by ethanol; hot-water extraction; alcohol sedimentation (glycoprotein); deproteinizing, which yielding 1.77% for 4.42g. Meanwhile, characteristics of Chestnut glycoprotein and Chestnut crude polysaccharide are mensurated, it shows that protein has been basically eliminated for there is no absorption peak at 280 nm.  Protein content of glycoprotein and crude polysaccharide are measured by Coomassie Brilliant Blue .which are separately to 2.28% and 0.026% . Hexose content of crude polysaccharide is measured by Phenol - sulfuric acid method to 89.54%. Infrared spectroscopy revealed that the crude polysaccharide may contain Beta-D-based glucopynanosyl.
 By Sephacryl S-100HR column and DEAE - Sepadax Fast Flow anion-exchange column, 8 components are separated from Chestnut crude polysaccharide. A single symmetrical peak is gained by Elution through Sephacryl S-100HR, what proves that the group is divided into a  homogeneous precision polysaccharide.
 Hydroxy free radical scavenging tests for chestnut crude polysaccharide is carried out using methyl violet model.It confirms that Chestnut crude polysaccharide has certain hydroxyl radical scavenging effect range 1-20mg/mL, and the effect begin strong as the bigger of the content of the crude polysaccharide. The scavenging activity on hydroxy free radical is weaker than Vc, the result shows, the scavenging effect of the crude polysaccharide (20mg/mL) is 34.8%, while Vc  (20mg/mL) is 66.08%.
Keywords：Chestnut; Polysaccharide; Extraction; Purification; Antioxidant Activity

目  录
中文摘要 I
英文摘要 II
目  录 III
1. 绪 论 1
1.1 板栗的研究概况 1
1.2 多糖的概述及其研究进展 2
2  实验部分 7
2.1 多糖的提取 7
2.2 板栗粗糖蛋白和粗多糖的理化性质测定 8
2.3 板栗粗多糖的分离纯化 10
2.4 板栗多糖的抗羟基自由基活性测试 11
3  结果与讨论 12
3.1板栗多糖的提取 12
3.2 板栗粗糖蛋白和粗多糖的理化性质测定 12
3.3 板栗粗多糖的分离纯化 17
3.4 板栗粗多糖对羟基自由基·OH的清除率测定 19
4. 总结与展望 21
致 谢 22
参考文献 23

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