【作者】 张武岗；

【导师】 周乐；

【作者基本信息】 西北农林科技大学 ， 植物学， 2006， 硕士

【摘要】 紫花地丁是堇菜科堇菜属植物,对其在医药临床方面的研究报道很多,而对其在抗菌成分方面的报道却很少,本文针对其抑菌活性成分进行了系统的研究。我们先对紫花地丁全草进行了系统的试管预试和薄层预试,初步确定出其中含有的主要化学成分:黄酮类、内酯香豆素类、挥发油、甾体萜类及少量有机酸和糖类。然后采用柱色谱分离纯化与抑菌活性测定跟踪相结合的研究方法来筛选活性成分。首先,将实验材料甲醇粗提液,依次用乙酸乙酯,氯仿,石油醚萃取,再以金黄色葡萄球菌（S.aureas）、无乳链球菌（S.agalactiae）、乳房链球菌（S.uberis）、停乳链球菌（S.dysagalactiae）、大肠杆菌（E.coli）和沙门氏菌（Salmonella）六种的菌种作示踪,再通过药敏片体外抑菌活性测定法,确定出其抑菌活性主要集中在醇水相和氯仿相,然后利用柱层析对这两部分进行分离纯化。醇水相依次经过丙酮沉淀、硅胶柱层析、结晶,得到纯品D₁;氯仿相依次经过硅胶柱层析、葡聚糖凝胶柱Sephadex LH-20,得到纯品D₂、D₄、D₅;再通过试管比浊法体外抑菌实验,确定出D₁、D₅的最低抑菌浓度和最低杀菌浓度,D₁、D₅的最小抑菌浓度基本相等,均可达到0.039mg/ml,但D₁、D₅的最低杀菌浓度有明显的差别,D₁的杀菌活性远远好于D₅,最低杀菌浓度分别为0.31mg/ml、2.5mg/ml。通过对四个化合物的理化定性及有机波谱解析,结合文献资料,推断出化合物D₁为七叶内酯（ aesculetin ）、化合物D₂为6、7-二甲氧基香豆素（ 6, 7-dimethoxycoumarin）、化合物D₄为东莨菪内酯（Scopoletin）、化合物D₅为6-羟甲基-7-甲氧基香豆素（6-hydroxymethyl-7-methoxycoumarin）,除化合物D₅为一个新化合物,其余均为已知化合物。更多还原

【Abstract】 Viola yedoensis Makino belongs to Viola. Although there are lots of reports about its application on medicine and clinical aspects recently, the research on the components of the antibacterial activities is little. The systematic research about its components on the antibacterial activities is involved in this thesis, which is on the basis of the predecessors’work.The chemical constituents of V. yedoensis Makino have been pretested systematically by means of tube-test and thin layer chromatography （TLC）. The main ingredients are flavonoids, coumarins, essential oils, steriod terpenoids, few organic acids and carbohydrates. We selected the active components from V. yedoensis Makino by column chromatography on silica gel, accompanied with the antibacterial activity tests.Frist, The coarse powder of Viola yedoensis Makino extracted with methanol was extracted with ethyl acetate, chloroform, petroleum ether. Then, the antibacterial activities of the extraction were tested by dish diffusion of Neo-Sensitab against S.aureas, S.agalactiae, S.uberis, S.dysagalactiae, E.col and Salmonella in vitro. The results showed that most of the antibacterial constituents in Viola yedoensis Makino existed in chloroform phase and ethanol-water（1:1） phase. These two phases were subjected to column chromatography.The compound D₁ was isolated from ethanol-water（1:1） phase by means of sedimentation in acetone, column chromatography on silica gel, crystallization. The compound D₂、D₄、D₅ were isolated from chloroform phase by means of column chromatography on silica gel and sephadex LH-20. Then, the minimal inhibitory concentrations（MIC） and minimal bactericidal concentrations（MBC） of these four compounds were tested by using nephelometric analysis. The results showed that the minimal inhibitory concentrations（MIC） of the compounds D₁ and D₅ were 0.039 mg/ml, however, there are obvious differences in their minimal bactericidal concentrations（MBC）. The bactericidal activities of compound D₁ were much better than D₅, and their minimal bactericidal concentrations（MBC） were 0.31 mg/ml,2.5 mg/ml respectively.Through physical and chemical quality analysis and organic spectrum determination,更多还原