迷果芹多糖提取分离纯化及其初步药理学活性研究

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	迷果芹多糖提取分离纯化及其初步药理学活性研究
其他题名	The study of polysaccharide’s extraction , separation and purification and preliminary research on pharmacological activity of Sphallerocarpus gracilis
	于瑞涛
学位类型	硕士
导师	陶燕铎
	2008-05-30
学位授予单位	中国科学院西北高原生物研究所
学位授予地点	西北高原生物研究所
关键词	迷果芹提取分离纯化药理活性
摘要	摘要迷果芹为伞形科迷果芹属植物。本论文以迷果芹的根为研究对象，对迷果芹多糖的提取，分离和纯化进行了研究。通过单因素实验和正交实验考察了提取温度、提取时间、提取次数和料液比对迷果芹多糖提取率的影响。采用苯酚硫酸法对迷果芹多糖进行了含量分析。通过DEAE-52柱层析和sephadexG-100对迷果芹多糖进行了分离和纯化。并对迷果芹粗多糖的药理学活性进行了初步的研究。试验结果如下： 1. 优化了迷果芹多糖的提取工艺参数，得到最佳提取条件。即提取温度90℃、提取时间2小时、提取次数为1次、料液比1:10。在此条件下的多糖得率为9.13%。 2.采用苯酚硫酸法对迷果芹粗多糖进行了含量分析, 在波长490nm处,以葡萄糖为对照品,得回归方程为: A=15.98381C-0.0796, r=0.9978 ,在0.0247～0.1235mg/mL范围内呈线形关系。通过样品含量测定可知，迷果芹中的原药材中的多糖含量为4.04%。迷果芹粗多糖醇沉1次、醇沉2次、醇沉3次和醇沉3次脱脂所测得的迷果芹粗多糖中的多糖含量分别为54.90%，53.40%，45.73%，50.90%。 3.迷果芹粗多糖经过脱脂,sevage法脱蛋白,2%双氧水脱色,再经过DEAE-52柱层析,得到四个多糖组分SGS-1,SGS-2,SGS-3,SGS-4。并对SGS-1和SGS-3上SephadexG-100柱进一步纯化。通过峰形检测了SGS-1和SGS-3纯度，得到了两个组分较为均一的多糖SGS-1和SGS-3。 4．通过迷果芹粗多糖（SGS）对荷瘤（肝癌H22）小鼠的影响，以及对小鼠方向辨别障碍的保护作用的研究，考察了迷果芹多糖的初步药理学活性。迷果芹多糖（SGS）对荷瘤（肝癌H22）小鼠瘤体重量无明显地影响，体重增长三个剂量组均明显高于阳性对照药环磷酰胺组。三个剂量组小鼠胸腺、脾指数均高于环磷酰胺组，但无统计学意义。迷果芹多糖提取物设计剂量对小鼠方向辨别障碍虽有一定的保护作用，但无统计学意义。
其他摘要	Abstract Sphallerocarpus gracilis belongs to Sphallerocarpus Bess of the family of Umbelllferae. This paper studied Sphallerocarpus gracilis polysaccharide’s extraction, separation and purification by taking the root of Sphallerocarpus gracilis as materials. A single factor and the orthogonal experiments were used to inspect the effect of the extraction temperature, the extraction time, frequency and solid-liquid ratio from Sphallerocarpus gracilis polysaccharide. Phenol-Sulferic acid method was used to determine the polysaccharides content of Sphallerocarpus gracilis. Through DEAE-52 column chromatography and sephadexG-100 was used to isolate and purify Sphallerocarpus gracilis polysaccharides. Sphallerocarpus gracilis Polysaccharide’s pharmacological activity was studied preliminarily. The results are as follows: 1. Optimize Sphallerocarpus gracilis polysaccharide’s extract technology. That is, temperature 90 ℃, extracting 1 time, each time 120min, solid-liquid ratio 1:10. Under these conditions the polysaccharides was got 9.13 gram in 100 gram from the root of Sphallerocarpus gracilis. 2. Phenol-sulfuric acid method was used on determination of content of Sphallerocarpus gracilis polysaccharide. The regression equation is: A = 15.98381C-0.0796, r = 0.9978. The result showed that a good linear relationship from 0.0247 mg / mL to 0.1235 mg / mL when the wavelength is 490 nm and glucose is the reference substance. The conten of polysaccharide of Sphallerocarpus gracilis is 4.04 %. The content of polysaccharide of Sphallerocarpus gracilis’s crude polysaccharide with Ethanol precipitation 1 times, Ethanol precipitation 2 times, Ethanol precipitation 3 times and Ethanol precipitation 3times gotten rid of fat is respectively 54.90 %, 53.40 %, 45.73 %, 50.90%. 3. Sphallerocarpus gracilis’s crude polysaccharide after degreasing, removing protein by the method of sevage and decolorization with 2% hydrogen peroxide, received four polysaccharide components SGS-1, SGS-2, SGS-3 , SGS-4 when DEAE-52 column chromatography was used. SGS-1 and SGS-3 was purified further by SephadexG-100. The peak shape showed that SGS-1 and SGS-3 are relatively homogeneous polysaccharides. 4. The impact to the tumor (Liver Cancer H22) of mice and the protective effect of identifying obstacles about direction on mice with Sphallerocarpus gracilis’s crude polysaccharide (SGS). The study inspected initial pharmacological activity of Sphallerocarpus gracilis polysaccharide. Sphallerocarpus gracilis’s crude polysaccharide (SGS) had no effect on the weight of tumor (Liver Cancer H22) of mice. Weight of mice in three-dose group is higher obviously than cyclophosphamide group that is the positive control drug. The thymus index and spleen index of three-dose group of Sphallerocarpus gracilis’s crude polysaccharide are higher than the cyclophosphamide group, but not statistically significant. Sphallerocarpus gracilis’s crude polysaccharide have a protective effect to a certain extent on identifying obstacles about direction on mice, but not statistically significant, too.
页数	58
语种	中文
文献类型	学位论文
条目标识符	http://210.75.249.4/handle/363003/3166
专题	中国科学院西北高原生物研究所
推荐引用方式 GB/T 7714	于瑞涛. 迷果芹多糖提取分离纯化及其初步药理学活性研究[D]. 西北高原生物研究所. 中国科学院西北高原生物研究所,2008.

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