牛心朴子生物碱的提取精制及其对尖孢镰刀菌的抑制活性初探

发布时间：2018-01-12 23:28

本文关键词：牛心朴子生物碱的提取精制及其对尖孢镰刀菌的抑制活性初探　出处：《北方民族大学》2017年硕士论文　论文类型：学位论文

【摘要】：枸杞作为宁夏经济作物的支柱,倍受政府的关注。但是,各种自然病菌、病虫害以及环境和人为等因素,直接或者间接地影响了宁夏枸杞的果实质量和产量,其中枸杞根腐病病菌对枸杞的影响最为严重。牛心朴子是生长于宁夏荒漠半荒漠地区的有毒植物,项目组前期的研究成果表明其生物碱类生成具有抗真菌的作用。本论文为了从牛心朴子中寻找能抑制尖孢镰刀菌的活性成分,对牛心朴子的生物碱进行了系统分离,在获得初步抑制活性的前提下,采用pH区带精制逆流色谱技术大量制备有效成分,对分离条件进行探讨。主要内容如下:根据项目组前期研究成果,论文以牛心朴子中的菲并吲哚里西丁类生物碱为分离目标,采用经典色谱方法和核磁共振技术,从牛心朴子总生物碱中分离了3个化合物,鉴定了两个化合物的结构,分别为7-demethoxylophorine(antofine)和一个脂链化合物,初步测定了鲜牛心朴子中生物碱的含量为1.63%。采用菌丝生长抑制法对总生物碱和antofine抑制尖孢镰刀菌的生长活性进行了初步研究结果表明,高浓度(40μg/mL和50μg/mL)下,总生物碱表现出与市售抗根腐病化学农药相近的抑制活性,提示有进一步开发为抗枸杞根腐病的植物源农药的潜力。值得注意的是,antofine在较低浓度(20μg/m L)下,就表现出良好的抑制尖孢镰刀菌生长的活性,且优于市售化学农药百菌清和多菌灵,antofine可以作为新的抗根腐病的植物源农药或先导化合物进行进一步的研究。pH区带精制逆流色谱是一种液液分配色谱。为了富集和纯化antofine,以正己烷-乙酸乙酯-甲醇-水为溶剂体系,以溶剂在仪器中的保留率和antofine的得率作为指标,筛选了保持剂的加入量、仪器转速、流动相流速等分离条件,优化出最佳分离条件为溶剂体系正己烷:乙酸乙酯:甲醇:水(3:6:1:5.5),仪器转速650r/min、流速2mL/min。以此条件,一次制备得到antofine 44mg,纯度90%。
[Abstract]:Lycium barbarum, as the pillar of economic crops in Ningxia, has attracted the attention of the government. However, various natural pathogens, pests and diseases, environmental and human factors, directly or indirectly affect the fruit quality and yield of Lycium barbarum in Ningxia. The pathogen of Lycium barbarum root rot has the most serious effect on Lycium barbarum. Prunus chinensis is a poisonous plant growing in the desert and semi-desert area of Ningxia. The previous research results of the project group showed that its alkaloids had antifungal effect. In order to search for the active components which could inhibit Fusarium oxysporum in this paper. The alkaloids of Prunus chinensis were systematically separated. On the premise of obtaining the primary inhibitory activity, a large number of effective components were prepared by using pH zone refining countercurrent chromatography technique. The main contents are as follows: according to the previous research results of the project group, the separation of phenanthrene indorisidine alkaloids from Prunus Niuxinensis was taken as the separation target. Three compounds were isolated from total alkaloids of Prunus chinensis by classical chromatography and nuclear magnetic resonance (NMR) technique, and the structures of two compounds were identified. They were 7-demethoxylophorine antofine) and a lipid chain compound. The content of alkaloid was 1.63. The activity of total alkaloids and antofine against Fusarium oxysporum was studied by hyphal growth inhibition method. . At high concentrations of 40 渭 g / mL and 50 渭 g / mL, the total alkaloids showed similar inhibitory activities as those of the marketed chemical pesticides against root rot. It is suggested that there is potential for further development of plant-based pesticides against Lycium barbarum root rot. It is worth noting that antofine is at a lower concentration of 20 渭 g / m ~ (L). It showed good activity in inhibiting the growth of Fusarium oxysporum and was superior to chlorothalonil and carbendazim in the market. Antofine can be further studied as a new phytochemical or lead compound against root rot. Ph zone refining countercurrent chromatography is a liquid-liquid partition chromatography. In order to enrich and purify anto. Fine. Using n-hexane ethyl acetate methanol-water as solvent system the retention rate of solvent in the instrument and the yield rate of antofine were used as indexes to select the amount of retention agent and the rotational speed of the instrument. The optimum separation conditions were as follows: solvent system: n-hexane: ethyl acetate: methanol: water: 3: 6: 1: 5.5. 5. The rotational speed of the instrument was 650 r / min. The flow rate was 2 mL / min. Under this condition, 44 mg antofine was prepared at one time, and the purity was 90 mg.
【学位授予单位】：北方民族大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S481

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