荒漠肉苁蓉资源的综合利用

发布时间：2018-07-24 16:06

【摘要】：荒漠肉苁蓉是一种有着长久应用历史的名贵中药材。目前,在荒漠肉苁蓉的培育、化学组分及药理活性方面的研究较为深入,但对荒漠肉苁蓉的产业化深加工方面的研究较少,限制了其应用范围和经济价值提升。论文以荒漠肉苁蓉的综合利用为目的,研究了荒漠肉苁蓉多糖提取、分离、结构修饰及生物活性。同时,实现了苯乙醇苷、甘露醇的分离纯化。研究结果可为荒漠肉苁蓉资源分级、综合利用,拓宽应用范围等奠定基础。论文取得的主要研究成果如下:(1)建立了一种新的荒漠肉苁蓉中单糖组分的分析方法。此方法可以同时检测多达11种单糖,包括葡萄糖、果糖、甘露醇、半乳糖、半乳糖醛酸、葡萄糖醛酸、阿拉伯糖、鼠李糖、核糖、岩藻糖和木糖。此方法有效地弥补了柱前衍生法检测单糖技术中,化学标记试剂无法修饰果糖及甘露醇的缺陷,同时避免了高效液相离子渗透色谱检测技术中峰形重叠的问题。在现有的技术条件下,能较快、较准确的得到荒漠肉苁蓉中所有的单糖的组成信息。另外,此法可以广泛应用于其它植物中单糖组成分析。(2)膜过滤分级分离纯化荒漠肉苁蓉多糖中聚合度不同的组分。得到3种分子量较均一的组分,依次命名为CDP-A、CDP-B、CDP-C,其分子量分别为4000 kDa、2400 kDa和1300 kDa。研究了在分级分离多糖过程中,不同过滤膜的污染机理。结果表明:微滤主要是膜孔堵塞;超滤是膜孔堵塞和膜孔缩小双重作用;纳滤是滤饼起主要作用。对3种不同聚合度的多糖进行单糖组分分析,结果表明,CDP-C含不饱和键的半乳糖醛酸较高,其抗氧化能力也较强。(3)建立了荒漠肉苁蓉中苯乙醇苷及甘露醇的高效分离技术。利用膜过滤多糖所得的透过液,通过大孔吸附树脂吸附苯乙醇苷,采用浓缩结晶法和乙醇溶液体系中热溶解-冷结晶法,得到高纯度的甘露醇。利用此技术,荒漠肉苁蓉提取物中甘露醇以及松果菊苷和毛蕊花糖苷(苯乙醇苷中的特征组分)的含量分别从12.10%,2.74%和 1.20%提高到 99.00%,23.45%和 9.31%。此外,与前期报道的大孔吸附树脂分离苯乙醇苷工艺相比较,本工艺中树脂的最大重复操作次数从5次提升至15次。(4)为了拓展荒漠肉苁蓉的应用领域,研究了荒漠肉苁蓉中的多糖CDP-C及其中的苯乙醇苷对酒精性肝损伤的抑制作用。通过体外细胞学实验、模型动物中血清学指标及肝组织指标的变化,研究了多糖CDP-C和苯乙醇苷的护肝效果。结果表明,荒漠肉苁蓉多糖CDP-C和苯乙醇苷均有很好的护肝作用。(5)通过硒化、硫酸化及乙酰化修饰,对荒漠肉苁蓉多糖CDP-B进行了改性。通过抗氧化性和体外抗肿瘤实验,比较了修饰前后荒漠肉苁蓉多糖的活性。结果表明,硒化多糖和硫酸化多糖的抗氧化能力和体外抗肿瘤活性有较明显的提升,但乙酰化修饰无明显变化。
[Abstract]:Cistanche deserticola is a valuable Chinese medicine with a long history of application. At present, the research on the cultivation, chemical composition and pharmacological activity of Cistanche deserticola is more thorough, but there are few studies on the industrial deep processing of Cistanche deserticola, which limits its application scope and economic value. The extraction, isolation, structure modification and biological activity of polysaccharides from Cistanche deserticola were studied in this paper. At the same time, the separation and purification of phenylethanoside and mannitol were realized. The results can lay a foundation for the classification, comprehensive utilization and application of Cistanche deserticola. The main results are as follows: (1) A new method for the analysis of monosaccharides in Cistanche deserticola was established. This method can be used to detect up to 11 monosaccharides including glucose fructose mannitol galacturonic acid glucuronic acid arabinose rhamnose ribose fucose and xylose. This method can effectively remedy the defect that chemical labeling reagent can not modify fructose and mannitol in the technique of precolumn derivatization, and avoid the problem of overlapping peaks in high performance liquid phase ion permeation chromatography (HPLC). Under the existing technical conditions, the information of all monosaccharides in Cistanche deserticola can be obtained quickly and accurately. In addition, this method can be widely used in the analysis of monosaccharide composition in other plants. (2) membrane filtration and fractionation were used to separate and purify the polysaccharides of Cistanche deserticola with different degree of polymerization. Three components with uniform molecular weight were obtained and named as CDP-An CDP-BU CDP-C, with molecular weights of 4000 kDa 2400 kDa and 1300 kDa, respectively. The fouling mechanism of different filtration membranes in the process of fractionation of polysaccharides was studied. The results show that microfiltration is mainly membrane pore blockage, ultrafiltration is membrane pore blockage and membrane pore reduction, and nanofiltration plays a major role in filter cake. Three kinds of polysaccharides with different degree of polymerization were analyzed for monosaccharide composition. The results showed that the galacturonic acid with unsaturated bond was higher and its antioxidant ability was stronger. (3) A high efficiency separation technique for phenylethanoside and mannitol in Cistanche deserticola was established. High purity mannitol was obtained by means of concentrated crystallization and thermal dissolution-cold crystallization of phenylethanol by macroporous resin adsorbing phenylethanol in the permeable solution of polysaccharide filtered by membrane. By using this technique, the contents of mannitol in the extract of Cistanche deserticola and the contents of pyranthrin and ruffin (the characteristic components of phenylethanoside) were increased from 12.1074% and 1.20% to 99.0023.45% and 9.31%, respectively. In addition, compared with the previously reported technology of separation of phenylethanoside with macroporous adsorption resin, the maximum repeated operation times of the resin were increased from 5 times to 15 times. (4) in order to expand the application field of Cistanche deserticola, The inhibitory effects of polysaccharide CDP-C and phenylethanoside on alcoholic liver injury in Cistanche deserticola were studied. The effects of polysaccharide CDP-C and phenylethanoside on protecting liver were studied by means of in vitro cytological experiments and the changes of serological indexes and liver tissue indexes in model animals. The results showed that the polysaccharides of Cistanche deserticola CDP-C and phenylethanoside had good liver protection effects. (5) the polysaccharide CDP-B of Cistanche deserticola was modified by selenization, sulfation and acetylation. The activities of polysaccharides of Cistanche deserticola before and after modification were compared by anti-oxidation and anti-tumor experiments in vitro. The results showed that the antioxidation ability and antitumor activity of selenium polysaccharide and sulfated polysaccharide were significantly improved, but the acetylation modification had no obvious change.
【学位授予单位】：中国科学院大学(中国科学院过程工程研究所)
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R284;R285

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