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摘要：鲍姆木层孔菌Phellinus baumii，是近年来开发的一种多年生药用真菌。桑黄具有很高的药用价值，桑黄中存在一些抗氧化，抗肿瘤和抗衰老等活性成分。为了进一步研究桑黄菌丝体中活性成分，本实验对一种新方法培养得到的桑黄菌丝体进行分离纯化。首先，按照极性的大小对桑黄菌丝体醇提物分别用石油醚，氯仿，乙酸乙酯和正丁醇四种有机溶剂进行萃取，对四个活性萃取相进行抗肿瘤活性实验，筛选出乙酸乙酯相是活性较好的萃取相，作为下一步分离纯化的材料。本毕业论文在传统的分离技术手段基础上，摸索高速逆流色谱的条件，达到快速精确制备的分离效果。通过高速逆流色谱和硅胶柱层析两种分类纯化方法配合，得到了四种化合物，分别是：原儿茶醛、柚皮苷、黄芩素和豆甾醇。为了进一步了解分离纯化得到的化合物的活性，本实验还对分离得到的4种化合物进行了抗肿瘤的活性研究。比较4种化合物之间的活性得到，样品3，即黄芩素的活性较好。20996
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关键词：桑黄菌丝体；高速逆流色谱；化合物；抗肿瘤活性
Isolation and Purification of the active components from Phellinus baumii mycelium by High-speed Counter-current Chromatography
Abstract: Phellinus baumii is a kind of old natural traditional Chinese medicine which dewelops well in recent years. Phellinus baumii has very high medicinal value,.There are some antioxidant anti-tumor and anti-aging active ingredients in Phellinus igniarius,. In order to further study the biological activity of Phellinus linteus mycelia, cultivation of Phellinus linteus mycelia obtained on a new method for the isolation and purification of the experiment. First, according to the polarity of the petroleum ether,chloroform, ethyl acetate and butyl alcohol, extraction of Phellinus linteus mycelia extract, on four active phase extraction for antitumor activity screening experiment, ethyl acetate extraction good active phase, as the next purification separation material. In this article, on the basis of traditional separation technology, exploration of high speed countercurrent chromatography condition, achieves the fast and effective separation and accurate preparation. With the high speed countercurrent chromatography and silica gel column chromatography purification method of two kinds of classification, four kinds of compounds were obtained,they are protocatechuic aldehyde, naringin, baicalein and stigmasterol. In order to further understand the purified compounds, 4 compounds of the experiments on the separation were studied antitumor activity. A comparison between these different compounds indicated that Sample 3，named Baicalein, showed a better biological activity.
KeyWord: Phellinus baumii mycelium; high-speed counter-current chromatography; component; anti-cancer activity
目录
1 绪论 1
1.1 桑黄研究概况 1
1.2 桑黄菌丝体研究概况 1
1.2.1 桑黄菌丝体中的化学成分 1
1.2.2 桑黄菌丝体的生物活性 2
1.3 高速逆流色谱 4
1.3.1 原理及特点 4
1.3.2 研究现状 4
1.4 研究目的与意义 4
1.5 主要研究内容 5
2 实验材料和方法 6
2.1 实验材料 6
2.1.1 供试菌株 6
2.1.2 试剂 6
2.1.3 实验材料 7
2.1.4 仪器与设备 7
2.2 实验方法 8
2.2.1 菌丝体醇提物制备 8
2.2.2 桑黄菌丝体醇提物不同萃取相的制备 8
2.2.4 桑黄菌丝体乙酸乙酯部分的柱层析分离纯化 11