臺灣博碩士論文加值系統

文旦（Citrus grandis (L.) Osbeck）為台灣重要經濟果樹之一。本研究以SPME結合GC及GC-MS鑑定新鮮文旦葉、花及果皮之揮發性成分。實驗結果顯示文旦葉之主要揮發性成分為sabinene及cis-β-ocimene；文旦花主要揮發性成分為linalool；而文旦果皮則以limonene為主要成分。本研究鑑定文旦盛花時期之盛開花朵與花苞，及文旦花之六個部位其揮發性成分差異，發現花朵、花苞、花瓣及花蕊之主要成分皆為linalool，此四個樣品之香氣成分相近；花托、子房、花蕊及柱頭所鑑定出之主要成分相同，為β-pinene、limonene、trans-β-ocimene及(E)-4,8-dimethyl-1,3,7-nonatriene。
本研究為了解新鮮樣品與加工後之差異，以水蒸氣蒸餾法萃取文旦之葉、花及果皮精油，其收率分別為0.50 ± 0.09 g/kg、0.86 ± 0.09 g/kg及19.90 ± 0.43 g/kg；以冷壓法萃取文旦果皮精油，收率為 7.93 ± 0.23 g/kg，並利用GC及GC-MS鑑定精油成分，結果顯示文旦葉精油主要成分為sabinene、β-pinene及trans-β-ocimene；文旦花精油主要成分為β-pinene、limonene及trans-β-ocimene；文旦果皮冷壓及水蒸氣蒸餾精油主要成分皆為β-pinene、myrcene及limonene。除此之外，本研究探討文旦精油對Staphylococcus aureus、Escherichia coli及Pseudomonas aeruginosa之抗菌能力，結果顯示，文旦葉精油之抑菌效果最佳；本實驗亦探討時間殺菌效力評估，發現精油之IC50濃度與殺菌時間並無直接關係。在抗氧化試驗上，結果顯示文旦葉精油之總酚含量最高，且清除DPPH自由基能力與螯合亞鐵離子之能力亦最佳，因此，深具研究開發之潛力。

Wentan pomelo (Citrus grandis (L.) Osbeck) is one of the most important economic fruit trees in Taiwan. Volatile compounds of leaves, flowers, and peels of Wentan pomelo has been studied by solid phase microextraction (SPME) coupled with GC and GC-MS in this study. The major constituents in leaves were sabinene and cis-β-ocimene; the dominated compound in flowers was linalool, and the main constituent in peels was limonene. Wentan pomelo flowers were distinguished to flower and buds, and they were separated by hands into six parts to analys volatile compounds of each part. The major constituent of whole flowers, flower buds, petals, and stamens were linalool, and other volatile compounds of the four parts were similar. The main volatile constituents of receptacles, ovaries, styles, and stigmas were about the same, the major compounds were β-pinene, limonene, trans-β-ocimene and (E)-4,8-dimethyl-1,3,7-nonatriene. In order to understand the differences between fresh samples and that after distillation, essential oils of Wentan leaves, flowers, and peels were obtained by steam distillation, yields were 0.50 ± 0.09 g•kg-1, 0.86 ± 0.09 g•kg-1 and 19.90 ± 0.43 g•kg-1, respectively. Cold-pressed peel essential oil yield was 7.93 ± 0.23 g•kg-1. The major component of Wentan pomelo leaf oil were sabinene, β-pinene and trans-β-ocimene ; the main component of wentan pomelo flower oil were β-pinene, limonene and trans-β-ocimene. The compounds of peel oil extracted from both cold pressed and steam distillation were limonene, β-pinene and myrcene. Essential oils were also testing their antioxidant activities using DPPH method. Wentan pomelo leaf oils showed containing a weak antioxidant and weak antimicrobial activities.

目錄
誌謝………………………..…………..……………………………...I
中文摘要………………………..…………..……………………………...II
英文摘要………………………..…………..…………………………….....III
目錄...…………………………………..………………………….………..IV
圖目錄………………………………..………….…..………..…………VII
表目錄……………………………………..………….……....…………VIII
附錄目錄………………………………..……..………….……..…...……IX
壹、前言…………………………………..………………...……..…………1
貳、文獻回顧………………………………..……………...……..…………3
　一、文旦………………………………..………………..……..…………3
　　　(一) 文旦簡介…………..………………..………………..…………3
　　　(二) 文旦之營養成分…………………………..………..…………4
　　　(三) 文旦之生物活性…………………………..………..…………4
　　　(四) 文旦精油相關文獻…………………………..………………..5
　二、精油………………………………..………………..………………..6
　　　(一) 揮發性成分萃取方式…………..…………..…………………..7
　　　(二) 精油之化學成分………………..………………..……………13
　　　(三) 精油應用……………………………………..………………..13
　三、抗菌活性之研究………………………………..………………….14
　　　(一) 精油抗菌活性成分…………..………………………..………14
　　　(二) 精油抗菌機制………………………………...…..…..….……14
　四、抗氧化活性之研究………………………………………..………15
　　　(一) 自由基與活性氧之種類與來源…………………………..…15
　　　(二) 自由基與活性氧對生物體之影響……….………………….15
　　　(三) 抗氧化劑作用機制………………………………….………16
　　　(四) 精油抗氧化活性之研究……………………………..………17
參、材料與方法………………………………………….………..………18
　一、研究設計………………………………..………………………….18
　二、實驗材料………………………………..…………..……..………19
　　　(一) 原料…………………………..……...……..……..…...………19
　　　(二) 藥品及試劑…………………………………………..………20
　　　(三) 培養基…………………………………………………..……21
　　　(四) 菌種………………………………………………..…………21
　三、實驗方法………………………………..…………………………22
　　　(一) 新鮮樣品揮發性成分分析…………………………..………22
　　　(二) 精油之萃取………………………………..…………………24
　　　(三) 直接注入法與頂空固相微萃取法之比較…………………..26
　　　(四) 揮發性成分分析與鑑定………………………….…………27
　　　(五) 文旦精油抗菌活性試驗……………………………………....28
　　　(六) 文旦精油抗氧化活性試驗………………………….………31
肆、結果與討論………………………………..………………………….33
　一、新鮮文旦之揮發性成分分析………………………….……..……33
　　　(一) 不同部位文旦之揮發性成分分析…………..……….……….33
　　　(二) 文旦花之細部揮發性成分分析……………….…..…………37
　二、精油成分分析……………….………………………...………43
　　　(一) 精油收率……………………………………………..………43
　　　(二) 文旦精油之揮發性成分……………………………..………45
　　(三) 比較冷壓法與水蒸氣蒸餾法萃取之文旦果皮精油之差異....55
　　(四) 比較新鮮文旦樣品與其精油之成分差異……………………59
　三、文旦精油之抗菌效果評估………………………………….……..…69
　　　(一) 試驗菌株及其培養…………………………………..………69
　　　(二) 文旦精油對不同致病菌之最小抑菌濃度測定............………69
　　　(三) 不同濃度文旦精油之抗菌效果……………………………….71
　　　(四) 時間殺菌效力評估試驗（Time kill assay）…………..………77
　四、文旦精油抗氧化活性……………………………………...………82
　　　(一) 文旦精油總酚含量測定……………………………..………..82
　　　(二) 文旦精油清除DPPH自由基能力………….……………….82
　 (三) 文精油螯合亞鐵離子能力…………………..…..……………83
(四) 抗氧化活性成分…………………………..…………………..83
伍、結論……………….…..………………………………….……..………88
陸、參考文獻…………………………………….………………..………90
柒、附錄…...…………………………………………….……..………101

 
圖目錄
圖1. 水蒸氣蒸餾法…….……………………………………...…………….8
圖2. 溶劑萃取法………………………………………..……………..…...10
圖3. 水蒸氣蒸餾-溶劑萃取法………………………………………..……10
圖4. 固相微萃取法…………………………………………………...…….12
圖5. 實驗流程………….………………………………...………..………..18
圖6. 採樣於台灣南投之文旦果園…. …………….……………….………19
圖7. 文旦葉…. …………………………………………………….…...…..23
圖8. 文旦花…. …………………………………………………..…..……..23
圖9. 文旦果皮…. …………………………………………………….……23圖10. 文旦花之不同部位…………………………….…………………….25
圖11. 甘蔗榨汁機……………………………………….………………….25
圖12. 不同萃取方法萃取文旦精油收率之比較…………………………..44
圖13. 新鮮文旦葉及文旦葉水蒸氣蒸餾精油之主要揮發性成分..........…62
圖14. 新鮮文旦花及水蒸氣蒸餾文旦花精油之主要揮發性成分………..65
圖15. 不同濃度之文旦精油對S. aureus之抑菌力分析…………….….....74
圖16. 不同濃度之文旦精油對E. coli之抑菌力分析…….…………….....75
圖17. 不同濃度之文旦精油對P. aeruginosa之抑菌力分析……….…….76
圖18. 文旦精油對S. aureus之時間殺菌效力評估………………….…….78
圖19. 文旦精油對E. coli之時間殺菌效力評估………………….……….79
圖20. 文旦精油對P. aeruginosa之時間殺菌效力評估…………...………80
圖21. 文旦精油之總酚含量………..………………………………………85
圖22. 文旦精油清除DPPH自由基能力之測定……..……………………86
圖23. 文旦精油螯合亞鐵離子能力………………………..………………87

表目錄
表1. 新鮮文旦不同部位之揮發性成分……………………………………35
表2. 文旦花之不同部位之揮發性成分……………………………………40
表3. 以直接注射法（DI）結合GC及GC-MS鑑定文旦精油之成分……..47
表4. 以頂空固相微萃取法（HS-SPME）結合GC及GC-MS鑑定文旦精油成分………………………………………..……………………...51
表5. 冷壓法與水蒸氣蒸餾法萃取之文旦果皮精油成分之比較.………. 56
表6. 新鮮文旦葉及其精油之揮發性化合物之比較……….…….……….60
表7. 新鮮文旦花及其精油之揮發性化合物比較…………....…………. .63
表8. 新鮮文旦果皮及其精油之揮發性化合物比較…………….………. 67
表9. 文旦精油對不同菌株最小抑菌濃度之比較…..…………………….70
表10. 文旦精油對不同菌株IC50之比較…...….………………..…………73
表11. 文旦精油之IC50與殺菌時間之比較……………………..…………81


 
附錄目錄
附錄1. 2010年台灣主要文旦產區之種植面積及產量…………………101
附錄2. 文旦之營養成分……………………………………………........102

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