【摘要】：荒漠肉蓯蓉是一種有著長(zhǎng)久應(yīng)用歷史的名貴中藥材。目前,在荒漠肉蓯蓉的培育、化學(xué)組分及藥理活性方面的研究較為深入,但對(duì)荒漠肉蓯蓉的產(chǎn)業(yè)化深加工方面的研究較少,限制了其應(yīng)用范圍和經(jīng)濟(jì)價(jià)值提升。論文以荒漠肉蓯蓉的綜合利用為目的,研究了荒漠肉蓯蓉多糖提取、分離、結(jié)構(gòu)修飾及生物活性。同時(shí),實(shí)現(xiàn)了苯乙醇苷、甘露醇的分離純化。研究結(jié)果可為荒漠肉蓯蓉資源分級(jí)、綜合利用,拓寬應(yīng)用范圍等奠定基礎(chǔ)。論文取得的主要研究成果如下:(1)建立了一種新的荒漠肉蓯蓉中單糖組分的分析方法。此方法可以同時(shí)檢測(cè)多達(dá)11種單糖,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、巖藻糖和木糖。此方法有效地彌補(bǔ)了柱前衍生法檢測(cè)單糖技術(shù)中,化學(xué)標(biāo)記試劑無法修飾果糖及甘露醇的缺陷,同時(shí)避免了高效液相離子滲透色譜檢測(cè)技術(shù)中峰形重疊的問題。在現(xiàn)有的技術(shù)條件下,能較快、較準(zhǔn)確的得到荒漠肉蓯蓉中所有的單糖的組成信息。另外,此法可以廣泛應(yīng)用于其它植物中單糖組成分析。(2)膜過濾分級(jí)分離純化荒漠肉蓯蓉多糖中聚合度不同的組分。得到3種分子量較均一的組分,依次命名為CDP-A、CDP-B、CDP-C,其分子量分別為4000 kDa、2400 kDa和1300 kDa。研究了在分級(jí)分離多糖過程中,不同過濾膜的污染機(jī)理。結(jié)果表明:微濾主要是膜孔堵塞;超濾是膜孔堵塞和膜孔縮小雙重作用;納濾是濾餅起主要作用。對(duì)3種不同聚合度的多糖進(jìn)行單糖組分分析,結(jié)果表明,CDP-C含不飽和鍵的半乳糖醛酸較高,其抗氧化能力也較強(qiáng)。(3)建立了荒漠肉蓯蓉中苯乙醇苷及甘露醇的高效分離技術(shù)。利用膜過濾多糖所得的透過液,通過大孔吸附樹脂吸附苯乙醇苷,采用濃縮結(jié)晶法和乙醇溶液體系中熱溶解-冷結(jié)晶法,得到高純度的甘露醇。利用此技術(shù),荒漠肉蓯蓉提取物中甘露醇以及松果菊苷和毛蕊花糖苷(苯乙醇苷中的特征組分)的含量分別從12.10%,2.74%和 1.20%提高到 99.00%,23.45%和 9.31%。此外,與前期報(bào)道的大孔吸附樹脂分離苯乙醇苷工藝相比較,本工藝中樹脂的最大重復(fù)操作次數(shù)從5次提升至15次。(4)為了拓展荒漠肉蓯蓉的應(yīng)用領(lǐng)域,研究了荒漠肉蓯蓉中的多糖CDP-C及其中的苯乙醇苷對(duì)酒精性肝損傷的抑制作用。通過體外細(xì)胞學(xué)實(shí)驗(yàn)、模型動(dòng)物中血清學(xué)指標(biāo)及肝組織指標(biāo)的變化,研究了多糖CDP-C和苯乙醇苷的護(hù)肝效果。結(jié)果表明,荒漠肉蓯蓉多糖CDP-C和苯乙醇苷均有很好的護(hù)肝作用。(5)通過硒化、硫酸化及乙酰化修飾,對(duì)荒漠肉蓯蓉多糖CDP-B進(jìn)行了改性。通過抗氧化性和體外抗腫瘤實(shí)驗(yàn),比較了修飾前后荒漠肉蓯蓉多糖的活性。結(jié)果表明,硒化多糖和硫酸化多糖的抗氧化能力和體外抗腫瘤活性有較明顯的提升,但乙�；揎棢o明顯變化。
[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.
【學(xué)位授予單位】：中國(guó)科學(xué)院大學(xué)(中國(guó)科學(xué)院過程工程研究所)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：R284;R285

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