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  • 學位論文

柳杉抽出成分對病媒蚊之毒殺與忌避活性

Mosquito Larvicidal and Repellent Activities of Extractives from Cryptomeria japonica

指導教授 : 張上鎮

摘要


病媒蚊會傳染多種疾病,如瘧疾、絲蟲病、黃熱病、登革熱及日本腦炎等,因此控制病媒蚊數量以減少疾病之傳播遂成為重要的課題。相較於合成化學藥劑,植物天然產物擁有毒性較低,較容易降解的優點。故本試驗評估黑心柳杉(Cryptomeria japonica D. Don)乙醇抽出物及其成分對埃及斑蚊(Aedes aegypti)及白線斑蚊(Aedes albopictus)第四齡幼蟲的抑制活性;此外,亦進行柳杉各部位精油及其成分進行埃及斑蚊及白線斑蚊之蚊蟲忌避試驗,了解各部位精油對蚊蟲之忌避潛能。 由抗病媒蚊幼蟲試驗結果得知,材部乙醇抽出物之正己烷分離部對埃及斑蚊及白線斑蚊幼蟲具有好抑制活性,進一步由正己烷分離部得到14個次分離部(H1~H14),其中,H5對病媒蚊幼蟲展現良好之抑制活性。續由H5分離鑑定得9種化合物,包括Ferruginol、epi-Cubenol、4-Acetoxycryptomerione、Sandaracopimaric acid、epi-Cubebol、Cubebol、β-Eudesmol、Isopimarol和Phyllocladan-16α-ol。比較Ferruginol、epi-Cubebol、Cubebol和Isopimarol 4種主成分之抗病媒蚊幼蟲活性,以epi-Cubebol與Cubebol之效果最佳。 另外,比較柳杉4種部位精油之忌避效果,結果發現葉子精油對埃及斑蚊與白線斑蚊具有最佳之活性,因此利用固相微萃取法(Solid phase microextraction, SPME)進行精油揮發成分之動態分析,藉此了解揮發成分與忌避活性間的關係,使用之萃取纖維為DVB/CAR/PDMS。試驗結果得知,忌避時間0 min時,空氣中精油之主要揮發成分為3-Carene、α-Terpinene、Limonene、γ-Terpinene與Terpinolene;當忌避時間延長為20 min、40 min及60 min,試驗箱中的精油主要揮發成分則為(-)-Terpinen-4-ol。因此,針對此6種成分進行蚊蟲忌避試驗,結果發現(-)-Terpinen-4-ol對埃及斑蚊及白線斑蚊之忌避效果最好。

並列摘要


Mosquito-transmitted diseases such as malaria, filariasis, yellow fever, dengue and Japanese encephalitis are very serious. Therefore, it is important to control mosquito vector population for preventing the spread of these diseases. Nature products offer an advantage over synthetic pesticides because of less toxic and easily biodegradable. The purposes of this study were to investigate the larvicidal activities of ethanolic extracts from Cryptomeria japonica against fourth-instar larvae of mosquitoes Aedes aegypti and Aedes albopictus and to evaluate the repellent activities of essential oil from C. japonica against adult of mosquitoes A. aegypti and A. albopictus. Results of mosquito larvicidal tests demonstrated that n-hexane soluble fraction of wood extract from C. japonica exhibited the best larvicidal activity against fourth-instar larvae of mosquitoes. Among fourteen subfractions (H1~H14) of n-hexane soluble fraction, H5 subfraction showed the best inhibitory activity against the mosquito larvae tested. In addition, nine compounds were isolated and identified from H5 subfraction, including ferruginol, epi-cubenol, 4-acetoxycryptomerione, sandaracopimaric acid, epi-cubebol, cubebol, β-eudesmol, isopimarol and phyllocladan-16α-ol. Comparison of the larvicidal activity of four major compounds (ferruginol, epi-cubebol, cubebol and isopimarol) revealed that cubebol and epi-cubebol exhibited the best activity against A. aegypti and A. albopictus. Comparison of repellent activity of essential oils from four plant parts of C. japonica revealed that leaf oil exhibited the best action against mosquitoes. In order to understand the relationship between volatile organic compounds and repellent activity, solid phase microextraction method (SPME) was employed to analyze volatile organic compounds of leaf oil. The SPME fiber used was coated with DVB/CAR/PDMS. The major volatile organic compounds in the box were 3-carene, α-terpinene, limonene, γ-terpinene and terpinolene at 0 min. Results of the tests demonstrated that the major volatile organic compound in the box was (-)-terpinen-4-ol at 20 min, 40 min and 60 min, respectively. Furthermore, the repellent activities of six compounds against adults of the mosquitoes were evaluated, results revealed that (-)-terpinen-4-ol exhibited the best repellent activity against A. aegypti and A. albopictus.

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