本文選題：透骨草 + 提取��； 參考：《西北農(nóng)林科技大學(xué)》2017年碩士論文

【摘要】：透骨草(Phryma leptostachya L.)是透骨草科透骨草屬的多年生草本植物,在東亞一直作為傳統(tǒng)的殺蟲(chóng)植物來(lái)使用。為了進(jìn)一步的研究和開(kāi)發(fā)這一殺蟲(chóng)植物,本論文以透骨草為研究對(duì)象,研究了其殺蟲(chóng)活性成分在植株不同部位的分布情況,探討了其提取工藝,對(duì)殺蟲(chóng)活性較高的部分進(jìn)行了分離純化并建立了快速分離高活性成分Haedoxan A的方法,并對(duì)分離得到的化合物進(jìn)行了殺蟲(chóng)活性評(píng)價(jià)。主要研究結(jié)果如下:1.針對(duì)透骨草植株的不同部位(根和莖葉),采用不同提取方法(冷浸法和超聲法)以及不同溶劑(石油醚、乙酸乙酯、甲醇)進(jìn)行提取。結(jié)果表明:透骨草殺蟲(chóng)活性成分主要分布在根中,而且處于中等偏小極性段中,乙酸乙酯部分最多。2.在活性追蹤指導(dǎo)下,使用不同的常見(jiàn)色譜分離手段(萃取、硅膠柱色譜、半制備高效液相色譜)和重結(jié)晶等方法對(duì)活性高的部分進(jìn)行分離純化,根據(jù)所得化合物的理化性質(zhì)和波譜數(shù)據(jù)(質(zhì)譜、1H-NMR、13C-NMR、DEPT、HSQC、HMBC、COSY等),對(duì)分離得到的單體化合物進(jìn)行結(jié)構(gòu)鑒定。一共分離鑒定了6個(gè)單體化合物,分別是透骨草靈I、透骨草靈II、雙氧木脂素A、雙氧木脂素E、豆甾醇、β-谷甾醇。3.建立了快速獲取高殺蟲(chóng)活性成分雙氧木脂素A的方法。將透骨草根用甲醇超聲提取后用乙酸乙酯萃取,用石油醚:乙酸乙酯=100:0-0:100進(jìn)行硅膠柱層析,利用TLC紫外顯色合并相似流分,用粘蟲(chóng)追蹤活性最高段組分,用石油醚:丙酮=3:1進(jìn)行硅膠柱層析,用粘蟲(chóng)(Mythimna separata)活性追蹤合并活性流分,用甲醇重結(jié)晶可直接得到雙氧木脂素A。4.對(duì)分離得到的化合物進(jìn)行了殺蟲(chóng)活性評(píng)價(jià)。分離得到的化合物主要對(duì)鱗翅目的昆蟲(chóng)具有較高活性且主要表現(xiàn)為胃毒活性,而對(duì)蘋(píng)果黃蚜(Aphis ciricola van der Goot)、溫室白粉虱(Trialeurodes vaporariorum)和酢漿草如葉螨(Tetranychina harti.Ewing)均無(wú)活性。其中透骨草靈I對(duì)粘蟲(chóng)(Mythimna separata)僅表現(xiàn)麻醉活性,對(duì)3齡粘蟲(chóng)4 h時(shí)的麻醉中濃度NC50為3450μg/mL,而對(duì)小菜蛾(Plutella xylostella)有毒殺活性,48 h胃毒致死中濃度為1432μg/mL。雙氧木脂素A對(duì)3齡粘蟲(chóng)胃毒活性致死中濃度LC50為17.06μg/mL,對(duì)3齡粘蟲(chóng)24 h觸殺LC50為1123μg/mL,對(duì)3齡小菜蛾幼蟲(chóng)48 h的致死中濃度LC50分別為857μg/mL。雙氧木脂素E對(duì)3齡粘蟲(chóng)胃毒活性致死中濃度LC50為60.51μg/mL,對(duì)3齡粘蟲(chóng)24 h觸殺LC50為3758μg/mL,對(duì)3齡小菜蛾幼蟲(chóng)48 h的致死中濃度LC50為1855μg/mL�；衔颰6對(duì)3齡粘蟲(chóng)8 h時(shí)的麻醉中濃度NC50為2807μg/mL,對(duì)3齡小菜蛾幼蟲(chóng)無(wú)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.
【學(xué)位授予單位】：西北農(nóng)林科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：TQ453

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