砂地柏枝叶中杀虫活性成分初步研究
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摘要
“西北农林科技大学无公害农药研究服务中心”首次发现并报道了砂地柏(Sabina vulgaris Ant.)的杀虫作用,并从其果实中分离得到了精油主要杀虫活性成分松油烯-4-醇和非精油杀虫活性成分脱氧鬼臼毒素,从其叶的较强极性部分中分离得到了活性成分鬼臼毒素,而未对砂地柏枝叶弱极性部分中活性成分进行分离。本研究以前期工作为基础,主要从砂地柏枝叶弱极性部分寻找其它杀虫活性成分,进行了以下几个方面的工作:
     活性成分的提取、分离与鉴定。砂地柏枝叶粉碎物以95%乙醇为溶剂,采用冷浸法提取,浸提液减压浓缩回收乙醇即得到乙醇浸膏。采用逆向分配法,将乙醇浸膏用去离子水饱和悬浮,依次用石油醚、氯仿、乙酸乙酯、正丁醇等不同极性的溶剂萃取,各萃取液浓缩后分别得到四种溶剂萃取物的浸膏。以粘虫(Mythimna separatea W.)3龄幼虫为试虫进行活性测试,结果表明:除正丁醇萃取物外,各段萃取物对试虫均有较高的拒食活性;氯仿和乙酸乙酯萃取物同时对试虫表现出了较强的毒杀活性。以粘虫3龄幼虫为试虫,采用生物活性跟踪法对石油醚萃取物进行了四次常压柱层析分离,每次均选取活性较高且样品量较大的馏分进行下一次分离,共分离得到了3个化合物:Y2、Y7、Y8。通过熔点测定和TLC检测,确定分离得到的化合物为纯化合物;通过测定紫外光谱(UV)、质谱(MS)、氢核磁共振谱(~1HNMR)数据,并与标准谱图相比较,鉴定出上述3个化合物分别为:β-谷甾醇(Y2)、二十二烷酸(Y7)、十六烷酸(Y8)。
     主要测定了β-谷甾醇对几种试虫的生物活性。结果表明:β-谷甾醇对菜青虫(Pieris rapae L.)5龄幼虫表现出较高的拒食活性,浓度为1mg/mL时,其48小时拒食率为61.57%;对粘虫3龄幼虫表现出一定的拒食活性,其AFC_(50)为8.859mg/mL;在供试剂量下,对白纹伊蚊(Aedes albopictus S.)幼虫和小菜蛾(Plutella xylostella L.)幼虫无明显生物活性。将β-谷甾醇对粘虫3龄幼虫的活性测定结果与乙醇提取物、石油醚萃取物及分离过程中各馏分对该试虫的活性结果相比较,推测其中可能还存在着其它活性成分,或存在着有互作关系的其它成分。
     通过本论文的研究,可以初步推测β-谷甾醇为砂地柏中活性成分之一。也同时说明,砂地柏杀虫活性成分比较复杂,开展进一步的活性成分研究及活性成分互作研究是十分必要的。
The insecticidal activity of Sabina vulgaris Ant. was fovmd and reported by the Research and Service Center of Biorational Pesticides, NWSUAF, subsequently, three insecticidal ingredients-tepineol-4-ol which is isolated from the essential oil, deoxypodophyllotoxin from the fruit and podophyllotoxin from the high polar extract of leaves, were isolated.The work of this treatise basing on the low polar extract of leaves, which is not studied before,looks for other insecticidal ingredients from Sabina vulgris Ant..The main results are as follows:
    The extraction, isolation and identification of the insecticidal constitution were studied. The powdered leaves of Sabina vulgaris Ant. was extracted with EtOH(95%), and the solvent was removed. The EtOH extract suspended by distilled water was allocated to four different polar solvent-Petroleum ether, Chloroform, Ethyl acetate and Butanol by Counter Current Distribution, the four extracts were concentrated by removing the solvent. The activity was tested by the larva of Mythimna separatea W.. The results idicated: all extracts ,except butanol extract, had better repellency active; in addition, chloroform and ethyl acetate abstracts had better toxicity. Mythimna separatea W. was used as test insect, and active guiding method was applied to test the activity of Petroleum ether abstracts. After four normal pressure pillar chromatographys, three active materials were gained, which were Y2, Y7, Y8. The melting point test and TLC test make clear that the active materials were pure counpond. Comparing to the standard chromatography chart, UV, MS, 1HNMR data make clear that the three materials were p-steriol (Y2), docosanoic acid(Y7), dioctyl acid(Y8).
    The insecticidal activity of p-steriol to several insects was investigated. The result showes it has higher antifeeding activity to Pieris rapae L.and the activity of 48h is 61.57%,with the dosage of Img/mL. It also has antifeeding activity to Mythimna separatea W. But it showes no any activity to Aedes albopictus S. and Plutella xylostella L.under tested dosage. Comparing to ethanol abstracts, pertroleum ether abstracts and various distillations, the p-steriol had different result. So some other ingredients may exist in Sabina vulgris Ant. or have interplay action with other materials.
    The paper told that the P-steriol was one of the active materials of Sabina vulgris Ant. and the ingredients of insecticide materials in Sabina vulgris Ant. were complex. So the further research on active materials and the interplay of which was very necessary.
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