臺灣博碩士論文加值系統

柑橘水果為台灣重要經濟作物之一，許多柑橘水果經加工後其果皮廢棄物數量極為龐大，柑橘果皮之油囊中富含精油及許多具保健功效之化合物，其中以Limonene含量最多。
一般民眾常於食用燒烤食品尤其是海鮮類之燒烤食品時，常塗佈檸檬汁以降低腥味與提升燒烤物風味。由於碳烤過程中產生多環芳香族碳氫化合物(Polycyclic Aromatic Hydrocarbons；PAHs)類的致癌物質，而柑橘類精油中富含具有抗致突變效果的terpene類物質(如Limonene) ，因此本研究擬探討海鮮類之燒烤食品表面塗佈檸檬汁液是否能夠降低食用碳烤食品所導致的致突變之風險。此外文獻指出，柑橘類皮中含有豐富的抗氧化及抗菌性物質，因此本研究擬以柑橘皮甲醇萃取物探討其抗氧化及抗菌效果，並探討將Limonene用於葡萄酒製程中取代二氧化硫，製作無二氧化硫葡萄酒之可行性。
實驗結果顯示，魚皮經210℃燒烤20分鐘後其PAHs含量顯著增加，利用Ame’s test檢測亦發現碳烤後的魚皮具有較高的致突變率。但燒烤物經Limonene, 檸檬果皮 Cold pressure oil 或水蒸氣蒸餾精油處理後，則致突變率顯著降低，顯示在食用燒烤類食物前，適量塗佈檸檬汁液有助於降低致突變及致癌的風險。此外，柑橘類果皮之甲醇抽出物對於 B. subtilis 及 S. aureus有很好的抗菌效果。若將Limonene添加於葡萄果醪後進行酒精發酵，發現所釀製之葡萄酒不論在酒精度、pH值等數據與添加二氧化硫組相近，但在總生菌數則遠低於二氧化硫組，顯示在無二氧化硫葡萄酒之製造上，Limonene相當具有潛力。綜合以上結果，柑橘果皮抽出物除了可改善燒烤食物本身的風味及降低pH以利於食物的保存外，研究成果顯示還可以降低致癌風險以及用於製造無二氧化硫葡萄酒。

Being one of major economic crops in Taiwan , the waste from citrus fruits was very tremendous mass. However, the peelings of citrus fruits are rich in essential oil, especially, the limonene are the major.To enrich the flavors and mask the undesirable flavors in grilled sea food meats, we often spread it with lemon juice. Whether the anti-carcinogenesis activities of terpene, such as limonene, in citrus fruits essential oil extraction can diminish the carcinogenesis risk of PAHs, from grilled sea food meats, was our present purpose.
In addition, it was well known that the peelings of citrus fruits are also rich in antioxidant and anti-bacterial activities. In present study, we also evaluated the feasible application of citrus fruits essential oil, especial the limonene, on anti-bacterial activities to substitute for the addition of sulfur dioxide. The PAHs contents in fish skin increased markedly after being grilled at 210℃ for 20 minutes. The greater carcinogenesis risk of grilled fish skin was observed by Ame’s test. The reduction of carcinogenesis risk of grilled fish skin, which is treated with limonene, cold pressure oil from lemon and extract from lemon peelings, indicated that the spreading of lemon juice can facilitate the diminishing carcinogenesis risk. The obviously antibiotics effects against the B. subtilis and S. aureus can be fond in citrus fruits essential oil extraction. The similar results of alcoholic content and pH value between the musts added limonene and sulfur dioxide has shown that substitution of limonene is potentially feasible to produce the sulfur dioxide free wines. Furthermore, the total count of bacteria in the musts added limonene was much lower than in sulfur dioxide. In summary, citrus fruits essential oil extraction can not only diminish the carcinogenesis risk of PAHs, from grilled sea food meats, but also can produce the sulfur dioxide free wines.

目錄

中文摘要……………………………………………………………………Ⅰ
英文摘要……………………………………………………………………Ⅲ
致謝………………………………………………………………………….Ⅴ
壹、文獻整理…………………………………………………………………1
一．PAHs的簡介……………………………………………………..…1
二．精油簡介…………………………………………………………10
三．葡萄酒簡介………………………………………………………24
四．抗氧化簡介………………………………………………………29
五．實驗架構………………………………………………………32

第一部份、Limonene及柑橘果皮抽出物之抗致突變性評估……………33
一．前言………………………………………………………………34
二．材料與方法………………………………………………………36
三．結果與討論………………………………………………………47
四．結論………………………………………………………………75

第貳部份、柑橘果皮萃取物及Limonene之生物活性並添加於葡萄釀製中
取代二氧化硫之可行性評估…………………………………76
一．前言………………………………………………………………77
二．材料與方法………………………………………………………79
三．結果與討論………………………………………………………88
四．結論………………………………………………………………128
貳、參考文獻………………………………………………………… 129




















表次

表1. 不同萃取方式之柑橘精油以GC-MS分析其揮發性化合物組成…52
表2. 柑橘精油對於鼠傷寒沙門氏菌株 TA100之毒性試驗……………53
表3. 柑橘精油對於鼠傷寒沙門氏菌株 TA100之致突變性試驗………55
表4. 柑橘精油對於鼠傷寒沙門氏菌株 TA100之抗致突變性試驗……57
表5. 燒烤魚皮萃取物對鼠傷寒沙門氏菌TA 100之毒性試驗…………..59
表6. 燒烤魚皮萃取物對傷寒沙門氏菌TA 100之致突變性試驗………61
表7. Limonene及檸檬精油針對燒烤魚皮萃取物 (200 μg/plate) 於鼠傷寒沙門氏菌TA 100之抗致突率……………………………………63
表8. Limonene及檸檬精油針對燒烤魚皮萃取物 (50 μg/plate) 於鼠傷寒沙門氏菌TA 100之抗致突變率……………………………………65
表9. 浸泡Limonene及檸檬精油之魚皮燒烤後萃取物對鼠傷寒沙門氏菌TA 100之毒性試驗…………………………………………………67
表10. 浸泡Limonene及檸檬精油之魚皮燒烤後在同一濃度下萃取物對
鼠傷寒沙門氏菌之致突變性試驗………………………………69
表11. 燒烤後浸泡Limonene及檸檬精油之魚皮萃取物對鼠傷寒沙門氏菌TA 100之毒性試驗…………………………………………………71
表12. 燒烤後浸泡Limonene及檸檬精油之魚皮萃取物對鼠傷寒沙門氏菌TA 100之致突變性試驗……………………………….……..73
表13. Limonene對七株病原菌之抑菌圈大小……………………………99
表14. 檸檬果皮甲醇萃取物對七株病原菌之抑菌圈大小……………100
表15. 晚崙西亞橙果皮甲醇萃取物對七株病原菌之抑菌圈大小………101
表16. 葡萄柚果皮甲醇萃取物對七株病原菌之抑菌圈大小…………102
表17. Limonene對於七株病原菌其抑制百分比……………………103
表18. 檸檬果皮之甲醇萃取物對於七株病原菌其抑制百分比………104
表19. 晚崙西亞橙果皮之甲醇萃取物對於七株病原菌其抑制百分…105
表20. 葡萄柚果皮之甲醇萃取物對於七株病原菌其抑制百分比……106
表21. Limonene及柑橘果皮甲醇萃取物對七株病原菌其最低抑制濃度及
最低殺菌濃度………………………………………………...……107
表22. 添加Limonene於葡萄釀造酒中其Lab值在不同發酵天數之影
響………………………………………………………………….115





圖次

圖1. 實驗架構圖……………………………………………………………32
圖2. 柑橘精油對於鼠傷寒沙門氏菌株 TA100之毒性試驗……………54
圖3. 柑橘精油對於鼠傷寒沙門氏菌株 TA100之致突變性試驗………56
圖4. 柑橘精油對於鼠傷寒沙門氏菌株 TA100之抗致突變性試驗……58
圖5. 燒烤魚皮萃取物對鼠傷寒沙門氏菌TA 100之毒性試驗…………..60
圖6. 燒烤魚皮萃取物對傷寒沙門氏菌TA 100之致突變性試驗………62
圖7. Limonene及檸檬精油針對燒烤魚皮萃取物 (200 μg/plate) 於鼠傷寒沙門氏菌TA 100之抗致突變率…………………………………64
圖8. Limonene及檸檬精油針對燒烤魚皮萃取物 (50 μg/plate) 於鼠傷寒沙門氏菌TA 100之抗致突變率……………………………………66
圖9. 浸泡Limonene及檸檬精油之魚皮燒烤後萃取物對鼠傷寒沙門氏菌TA 100之毒性試驗…………………………………………………68
圖10. 浸泡Limonene及檸檬精油之魚皮燒烤後在同一濃度下萃取物對鼠傷寒沙門氏菌之致突變性試驗…………………………………70
圖11. 燒烤後浸泡Limonene及檸檬精油之魚皮萃取物對鼠傷寒沙門氏菌TA 100之毒性試驗…………………………………………………72
圖12. 燒烤後浸泡Limonene及檸檬精油之魚皮萃取物對鼠傷寒沙門氏菌TA 100之致突變性試驗………………………………….……..74
圖13. 三種柑橘果皮甲醇萃取物在不同濃度下其自由基清除能力之影
響…………………………………………………………………108
圖14. 三種柑橘果皮甲醇萃取物在不同濃度下其還原能力之影響…109
圖15. 添加Limonene於葡萄釀造酒中其乙醇濃度在不同發酵天數之
影響………………………………………………………………110
圖16. 添加Limonene於葡萄釀造酒中其可溶性固形物在不同發酵天
數之影響…………………………………………………………111
圖17. 添加Limonene於葡萄釀造酒中其pH值在不同發酵天數之
影響………………………………………………………………112
圖18. 添加Limonene於葡萄釀造酒中其可滴定酸百分比在不同發酵
天數之影響………………………………………………………113
圖19. 添加Limonene於葡萄釀造酒中其透光率在不同發酵天數之影
響…………………………………………………………………114
圖20. 添加Limonene於葡萄釀造酒中其褐變度在不同發酵天數下之影
響…………………………………………………………………116
圖21. 添加Limonene於葡萄釀造酒中其OD 620nm在不同發酵天數下
之影響……………………………………………………………117
圖22. 添加Limonene於果醪對於發酵後葡萄酒總酚含量之影響……118
圖23. 添加Limonene於果醪對於發酵後葡萄總色素含量評之影響…119
圖24. 添加Limonene於果醪對於發酵後葡萄酒類黃酮含量之影響…120
圖25. 添加Limonene於果醪對於發酵後葡萄酒蘋果酸含量之影響…121
圖26. 添加Limonene於果醪對於發酵後葡萄酒乳酸含量評之影響…122
圖27. 添加Limonene於果醪對於發酵後葡萄酒黏度之影響…………123
圖28. 添加Limonene於果醪對於發酵後葡萄酒自由基清除能力之影響…………………………………………………………………124

圖29. 添加Limonene於果醪對於發酵後葡萄酒三氯價鐵還原能力之影響…………………………………………………………………125
圖30. 添加Limonene於葡萄釀造酒中其總生菌數在不同發酵天數之影
響…………………………………………………………………126
圖31. 添加Limonene於果醪對於發酵後葡萄酒感官品評之影響……127

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