【作者】 陈鹏；

【导师】 潘远江；

【作者基本信息】 浙江大学 ， 化学， 2014， 硕士

【摘要】 复杂天然产物中活性有机物分离分析方法的研究是当代有机分析化学的重要内容,也是药物研发中寻找先导化合物的重要环节。高速逆流色谱作为一种无需固体载体的分离技术,已广泛应用于天然产物中活性成分的分离纯化。因此,开展逆流色谱分离天然产物中活性成分的相关研究有着重要的科学意义和应用价值。围绕这一目标,本论文具体开展了以下工作：1.活性导向的高速逆流色谱技术分离纯化异叶蛇葡萄中的抗氧化成分。首先进行在线活性测试,将异叶蛇葡萄粗品与羟基自由基在线反应,筛选出5个具有较好活性的化合物。选取上述5个化合物为分离目标,采用溶剂系统甲基叔丁基醚／乙腈／水(2/2/3,v/v)及环己烷／甲基叔丁基醚／乙酸乙酯／水(1/4/3/5,v/v),从100mg的粗品中成功分离出5个成分。经结构鉴定,这5个化合物分别为：金丝桃苷(1),异槲皮苷(2),芦丁(3),山柰酚-3-0-芸香糖苷(4)及槲皮素(5)。活性测试表明上述5个化合物均具有较好清除DPPH和羟基自由基的能力,其IC50值分别为18.2±1.3,17.0±1.4,24.2±1.2,38.1±1.7,9.0±1.2μM。抗氧化剂对羟基自由基的清除率(SRA)为65±5%,68±4%,96±2%,70±4%,98±2%。上述结果显示,使用该策略进行分离,可以大大提高活性化合物的纯化效率,在天然产物的活性成分的分离纯化中具有良好应用前景。2.利用逆流色谱分离得到杭白菊粗提物中的目标活性化合物。粗提物经LC/MS/MS分析及与标准品对照,确认其主要成分为活性化合物芹黄素-5-O-葡萄糖苷和金合欢素-7-O-葡萄糖苷,随后选取甲基叔丁基醚/正丁醇/乙腈/水(1.5/2/1/5,v/v)为溶剂体系进行逆流色谱分离,得到上述两化合物的纯品。实验表明,逆流色谱是一种制备高纯度活性成分的有效手段。此外,论文就高速逆流色谱近些年的方法发展作了简要总结。更多还原

【Abstract】 Separation and purification of bioactive natural products is important in organic analytical chemistry and drug discovery. As a liquid-liquid chromatography, Countercurrent chromatography (CCC) has widely been used in the separation of natural products. Therefore, methodology study for separation of bioactive natural products via countercurrent chromatography is not only meaningful in science but also valuable in economy. The specific research works were listed as follows:In the first part, the bioassay guided separation of antioxidants from Ampelopsis heterophylla was carried out via CCC. The antioxidant activity of the crude extract was initially evaluated by on-line HPLC method. Five compounds in the crude extract, namely hyperoside (1), isoquercitrin (2), rutin (3), kaempferol-3-rutinoside (4) and quercetin (5), exhibited good antioxidant activities. These compounds were further separated by countercurrent chromatography using biphasic solvent systems. Their structures were then identified by mass spectrometry and NMR. All the compounds exhibited significant DPPH radical scavenging activities with the IC50values at18.2±1.3,17.0±1.4,24.2±1.2,38.1±1.7and9.0±1.2/M, respectively. The scavenging ratios of the compounds against hydroxyl radicals were65±5%,68±4%,96±2%,70±4%and98±2%, respectively. This strategy could improve the separation efficiency and may have promising application.In the second part, two target bioactive compounds, namely apigenin-7-O-glucoside and acacia-7-O-glucoside, were separated from Chrysanthemum morifolium via countercurrent chromatography with the solvent system MtBE/n-Butanol/ACN/H2O(1.5/2/1/5, v/v).Moreover, the recent progress of countercurrent chromatography, including the elution modes, is also reviewed in this paper.更多还原