中药茜草乙酸乙酯提取物的分离及其抗氧化性活性研究.doc

摘 要 茜草是我国一种传统中药，其化学成分以蒽醌及其糖苷类化合物为主，具有抗氧化、抗菌、抗炎、抗肿瘤、抑制人体表皮细胞增殖等生物活性，可作为天然抗氧化剂治疗由自由基引起的疾病，并且无毒副作用。 本实验结合吸附柱色谱法和薄层色谱法，辅助于结晶、重结晶等方法分离中药茜草乙酸乙酯萃取物中的抗氧化活性成分。 基于已有实验基础，选取低极性段冲洗下来的ⅡL系列物质为研究对象，以环己烷：乙酸乙酯作为洗脱体系进行进一步分离，经两次柱层析分离，得到较纯组分ⅣLBB。ⅣLBB组分通过结晶、重结晶进一步纯化得到单体化合物ⅡLBB-1；采用铁离子还原（FRAP) 法及二苯代苦味酰基自由基（DPPH) 法对ⅡL，ⅡLB，ⅡLBB-1 的总抗氧化活性及清除自由基活性进行了测定。ⅣLBB-1经薄层色谱检测只有一个斑点，说明ⅡLBB-1组分是单体化合物；ⅡLBB-1的抗氧化活性高于ⅡL但低于ⅡLB，且都低于Vc、Ve。 柱层析；分离；抗氧化 Abstract Rubia Cordifolia L. is a traditional medicine of China. The main chemical composition is anthraquinones and its glycosides .It has antioxidant, antibacterial, anti-inflammatory, anti-tumor, inhibition of human epidermal cell proliferation and other biological activity, can be treated diseases caused by free radical as a natural anti-oxidant , and has no side effects.  This study combined adsorption column chromatography and thin layer chromatography (TLC) to separate the antioxidant components from ethyl acetate extract of Chinese medicine Rubia Cordifolia L. Based on the original experiment, select ⅡL which was seperated from the section of low polar as the research object for further purification. Select the cyclohexane: ethyl acetate as elution system, after twice seperation, we obtain ⅡLBB fraction, then recrystallization, re-crystallized, ⅡLBB is obtained, TLC showed only one spot. So fraction ⅡLBB-1 is pure compound. The total antioxidant activity, scavenging of free radical activity of ⅡL,ⅡLB,ⅡLBB-1were evaluated by the iron reduction (FRAP) method and 1, 1-pheny-2-picrylhydrazyl(DPPH) method, use Vc, Ve as positive control. The results showed that the antioxidant activity of ⅡLBB-1 is higher than ⅡL but less than ⅡLB, and all lower than Vc, Ve. Keywords: Rubia Cordifolia L; free radicals; chromatography; antioxidant  目 录 摘 要 I Abstract II 第1章 文献综述 1 1.1 药用植物茜草 1 1.1.1 茜草简介 1 1.1.2 茜草的主要化学成分 1 1.1.3 茜草的药理作用 1 1.2 分离方法 4 1.2.1 结晶、重结晶法 4 1.2.2 色谱法 4 1.3抗氧化活性测定 8 1.3.1 Fe3+ 还原 (FRAP) 法 8 1.3.2 硫代巴比妥酸 (TBA) 法 8 1.3.3 二苯代苦味酰基自由基 (DPPH