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研究生:古惠菁
研究生(外文):Hui-Jing Gu
論文名稱:柳杉抽出成分對病媒蚊之毒殺與忌避活性
論文名稱(外文):Mosquito Larvicidal and Repellent Activities of Extractives from Cryptomeria japonica
指導教授:張上鎮張上鎮引用關係
指導教授(外文):Shang-Tzen Chang
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:森林環境暨資源學研究所
學門:農業科學學門
學類:林業學類
論文種類:學術論文
論文出版年:2008
畢業學年度:97
語文別:中文
論文頁數:108
中文關鍵詞:埃及斑蚊白線斑蚊黑心柳杉抗病媒蚊幼蟲活性忌避活性
外文關鍵詞:Aedes aegyptiAedes albopictusBlack heartwood-type Cryptomeria japonicaLarvicidal activityRepellent activity
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  • 被引用被引用:2
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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。
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.
目錄 I
表目錄 IV
圖目錄 VI
摘要 XII
Abstract XIV
壹、前言 1
貳、文獻回顧 4
一、殺蚊幼蟲活性 4
(一)林木精油及抽出物之抗病媒蚊幼蟲活性 4
(二)天然抽出成分之抗病媒蚊幼蟲活性 7
二、蚊蟲忌避活性 10
(一)林木精油及抽出物之忌避活性 11
(二)天然抽出成分之忌避活性 14
三、蚊蟲忌避試驗 17
(一)人體忌避試驗 17
1. 試驗地點及時間 17
2. 試驗部位 19
3. 試樣之溶劑 19
(二)忌避裝置試驗 19
1. 忌避裝置A 20
2. 忌避裝置B 21
3. 忌避裝置C 22
4. 忌避裝置D 23
(三)忌避活性之評估方法 25
四、林木揮發成分之測定 25
五、柳杉之生物活性 28
參、材料與方法 37
一、試驗材料 37
(一)柳杉 37
(二)病媒蚊 37
二、試驗方法 37
(一)精油萃取 37
(二)精油成分鑑定 38
(三)抽出物萃取 38
(四)抽出成分的分離與純化 39
(五)化合物結構鑑定 39
(六)抗病媒蚊幼蟲活性 40
(七)病媒蚊成蟲之忌避活性 40
﹙八﹚頂空取樣法測定精油揮發成分 41
﹙九﹚SPME吸附法測定精油揮發成分 42
1. SPME 吸附纖維選擇 42
2. SPME吸附條件之選擇 43
3. 不同時間點精油揮發成分之測定 43
(十)統計分析 44
肆、結果與討論 45
一、柳杉抽出成分之抗病媒蚊幼蟲活性 45
(一)柳杉各部位乙醇抽出物之抗病媒蚊幼蟲活性 45
(二)柳杉材部乙醇抽出物4個可溶部之抗病媒蚊幼蟲活性 48
(三)正己烷可溶部14個次分離部之抗病媒蚊幼蟲活性 51
(四)柳杉抽出成分之分離與鑑定 52
1. 化合物S1之分離與鑑定 53
2. 化合物S2之分離與鑑定 56
3. 化合物S3之分離與鑑定 58
4. 化合物S4之分離與鑑定 60
5. 化合物S5之分離與鑑定 62
6. 化合物D1之分離與鑑定 64
7. 化合物D2之分離與鑑定 66
8. 化合物D3之分離與鑑定 68
9. 化合物D4之分離與鑑定 70
(五)柳杉抽出成分之抗病媒蚊幼蟲活性 72
二、柳杉精油之成蚊忌避活性 73
﹙一﹚不同部位精油之成蚊忌避活性 73
(二)精油揮發成分之分析 77
﹙三﹚SPME吸附纖維萃取效率之比較 82
﹙三﹚SPME對柳杉精油揮發成分之最佳萃取條件 87
﹙四﹚有效精油之成分分析 89
﹙五﹚葉子精油主成分之成蚊忌避活性 91
伍、結論 96
陸、參考文獻 98
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