透骨草杀虫成分提取方法及分离研究

发布时间：2018-06-10 12:34

本文选题：透骨草 + 提取　；参考：《西北农林科技大学》2017年硕士论文

【摘要】：透骨草(Phryma leptostachya L.)是透骨草科透骨草属的多年生草本植物,在东亚一直作为传统的杀虫植物来使用。为了进一步的研究和开发这一杀虫植物,本论文以透骨草为研究对象,研究了其杀虫活性成分在植株不同部位的分布情况,探讨了其提取工艺,对杀虫活性较高的部分进行了分离纯化并建立了快速分离高活性成分Haedoxan A的方法,并对分离得到的化合物进行了杀虫活性评价。主要研究结果如下:1.针对透骨草植株的不同部位(根和茎叶),采用不同提取方法(冷浸法和超声法)以及不同溶剂(石油醚、乙酸乙酯、甲醇)进行提取。结果表明:透骨草杀虫活性成分主要分布在根中,而且处于中等偏小极性段中,乙酸乙酯部分最多。2.在活性追踪指导下,使用不同的常见色谱分离手段(萃取、硅胶柱色谱、半制备高效液相色谱)和重结晶等方法对活性高的部分进行分离纯化,根据所得化合物的理化性质和波谱数据(质谱、1H-NMR、13C-NMR、DEPT、HSQC、HMBC、COSY等),对分离得到的单体化合物进行结构鉴定。一共分离鉴定了6个单体化合物,分别是透骨草灵I、透骨草灵II、双氧木脂素A、双氧木脂素E、豆甾醇、β-谷甾醇。3.建立了快速获取高杀虫活性成分双氧木脂素A的方法。将透骨草根用甲醇超声提取后用乙酸乙酯萃取,用石油醚:乙酸乙酯=100:0-0:100进行硅胶柱层析,利用TLC紫外显色合并相似流分,用粘虫追踪活性最高段组分,用石油醚:丙酮=3:1进行硅胶柱层析,用粘虫(Mythimna separata)活性追踪合并活性流分,用甲醇重结晶可直接得到双氧木脂素A。4.对分离得到的化合物进行了杀虫活性评价。分离得到的化合物主要对鳞翅目的昆虫具有较高活性且主要表现为胃毒活性,而对苹果黄蚜(Aphis ciricola van der Goot)、温室白粉虱(Trialeurodes vaporariorum)和酢浆草如叶螨(Tetranychina harti.Ewing)均无活性。其中透骨草灵I对粘虫(Mythimna separata)仅表现麻醉活性,对3龄粘虫4 h时的麻醉中浓度NC50为3450μg/mL,而对小菜蛾(Plutella xylostella)有毒杀活性,48 h胃毒致死中浓度为1432μg/mL。双氧木脂素A对3龄粘虫胃毒活性致死中浓度LC50为17.06μg/mL,对3龄粘虫24 h触杀LC50为1123μg/mL,对3龄小菜蛾幼虫48 h的致死中浓度LC50分别为857μg/mL。双氧木脂素E对3龄粘虫胃毒活性致死中浓度LC50为60.51μg/mL,对3龄粘虫24 h触杀LC50为3758μg/mL,对3龄小菜蛾幼虫48 h的致死中浓度LC50为1855μg/mL。化合物T6对3龄粘虫8 h时的麻醉中浓度NC50为2807μg/mL,对3龄小菜蛾幼虫无活性。
[Abstract]:Phryma leptostachya L. It is a perennial herb of the genus Osteopodiaceae, which has been used as a traditional insecticidal plant in East Asia. In order to further study and develop this insecticidal plant, the distribution of its insecticidal active components in different parts of the plant was studied in this paper, and the extraction process was discussed. The high insecticidal component Haedoxan A was isolated and purified, and the insecticidal activity of the isolated compounds was evaluated. The main results are as follows: 1. Different extraction methods (cold immersion method and ultrasonic method) and different solvents (petroleum ether ethyl acetate methanol) were used to extract different parts of the plant (root and stem and leaf). The results showed that the insecticidal components were mainly distributed in the roots, and were in the middle and small polar segments, and the ethyl acetate part was the most. 2. Under the guidance of activity tracing, the highly active parts were separated and purified by different chromatographic methods (extraction, silica gel column chromatography, semi-preparative high performance liquid chromatography) and recrystallization. Based on the physicochemical properties of the compounds and the spectral data (1H-NMR-13C-NMR-DEPTT-HSQC- HMBCCOSY et al.), the structures of the isolated monomers were identified. A total of six monomers were isolated and identified, namely, Tylophorin I, Tylophorin II, Dioxyllignin A, Dioxyllignin E, Soysterol, 尾 -sitosterol, and 尾 -sitosterol, respectively. A rapid method for obtaining dioxyllignin A with high insecticidal activity was established. The root was extracted with ethyl acetate after ultrasonic extraction with methanol. The silica gel column chromatography was carried out with petroleum ether: ethyl acetate 100: 0-0: 100. TLC UV coloration combined with similar flow fraction was used to trace the most active components. Petroleum ether: acetone 3: 1 was used for silica gel column chromatography. Dioxyllignin A.4was obtained by methanol recrystallization by tracing the activity of Mythimna separata. The insecticidal activity of the isolated compounds was evaluated. The compounds mainly showed high activity to Lepidoptera insects and showed gastric toxicity, but not to Aphis ciricola van der Goot, Trialeurodes vaporariorum and Tetranychina harti.Ewing. The concentration of NC50 was 3450 渭 g / mL for the 3rd instar Mythimna separata, and 1432 渭 g / mL for the diamondback moth Plutella xylostella. the concentration of NC50 was 1432 渭 g / mL for 48 h. The median lethal concentration (LC50) of xyloxylignin A was 17.06 渭 g / mL for the third instar armyworm, 1123 渭 g / mL for the 3rd instar moth for 24 h, and 857 渭 g / mL for 48h for the third instar diamondback moth larvae. The median lethal concentration (LC50) of xyloxylignin E was 60.51 渭 g / mL for the third instar armyworm, 3758 渭 g / mL for the 3rd instar armyworm for 24 h, and 1855 渭 g / mL for 48h for the third instar diamondback moth larvae. The average anesthetic concentration of compound T6 was 2807 渭 g 路mL ~ (-1) against the 3rd instar Trichinella xylostella larvae at 8 h, but it was not active to the 3rd instar diamondback moth larvae.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：TQ453

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