臺灣博碩士論文加值系統

本研究以成熟的金桔為原料，分別利用壓榨法、蒸氣蒸餾法、減壓蒸氣蒸餾法作金桔精油之萃取。以三種萃取法之比較與個別適用範圍作研究；以萃取量與萃取物品質來比較壓榨法、蒸氣蒸餾法與減壓蒸氣蒸餾法之優劣。
另將萃取的金桔精油進行生物活性研究，採抗氧化活性(清除DPPH自由基能力)及抗菌性試驗。試驗菌有大腸桿菌( Escherichia coli )、綠膿桿菌( Pseudomonas aeruginosa )、糞便腸球菌( Enterococcus faecalis )三種菌抑菌測試。最後以氣相層析質譜儀( GC-MS )與核磁共振儀( NMR )，作分析金桔精油主要組成份確認。
結果發現，以萃取量與萃取品質而言，以減壓蒸汽蒸餾法萃取為宜；以2.0％金桔精油濃度對清除DPPH自由基最佳(31.0％)；抑菌試驗的結果顯示2.0％金桔精油對糞便腸球菌抑菌效果最佳(抑菌環直徑15.8 mm)；其次為1.5％金桔精油對綠膿桿菌抑菌效果(抑菌環直徑13.7 mm)；但2.0％金桔精油對大腸桿菌抑菌效果仍屬為中度活性(抑菌環直徑12.8 mm)。最後金桔精油之主要成分經確認為檸檬油精(limonene)。

This investigation used the fully ripened Citrus microcarpa Bonge as the raw material for essential oil extraction. Extracts of essential oil from the peels were conducted expression, steam evaporating, and reduced pressure steam distillation, respectively. Yield and the quality of the extracts were studied to evaluate these three methods.
Bioactivities including DPPH scavenging capability and bacteriostasis were conducted on the extracted essential oils. Bacteriostatic ability were performed on Escherichia coli, Pseudomonas aeruginosa, and Enterococcus faecalis. Finally, GC-MS, and NMR were used to confirmed the primary composition of essential oils from Citrus microcarpa Bonge.
The results verified that reduced pressure steam distillation is the best extraction method among these three methods applied, judging by the yield and the quality of essential oils. The best DPPH scavenging ability was found with 2.0% of essential oil (31.0%). The data from bacteriostasis showed that 2.0% of essential oil had the best inhibition on Enterococcus faecalis (diameter of bacteriostatic ring was 15.8 mm). What came next was that 1.5% of essential oil on Pseudomonas aeruginosa (ring diameter was 13.7 mm). The inhibition of 2.0% essential oil on Escherichia coli was found to be moderate (ring diameter was 12.8 mm). Finally the primary composition of Citrus microcarpa Bonge essential oil was confirmed as limonene.

摘要···················································· Ⅰ
Abstract ················································ Ⅲ
誌謝····················································· Ⅴ
目錄····················································· Ⅵ
圖表目錄·················································· Ⅸ
第1 章前言················································· 1
第2 章文獻回顧············································· 2
2.1 金桔·················································· 2
2.2 香氣化合物············································ 2
2.2.1 香氣化合物的生合成································· 2
2.2.2 香氣的主要化合物··································· 3
2.3 天然植物精油·········································· 3
2.3.1 天然植物精油萃取方法······························ 3
2.3.2 植物精油的化學介紹································ 5
2.3.3 植物精油在人體的傳達途徑·························· 6
2.4 抗氧化活性··········································· 11
2.4.1 自由基的定義····································· 11
2.4.2 活性氧的定義····································· 11
2.4.3 柑橘類的抗氧化活性······························· 11
2.5 抗菌活性之研究······································· 13
2.5.1 精油之抗菌活性評估法····························· 13
2.5.2 精油成份與抗菌性之相關··························· 13
2.5.3 三種試驗菌介紹··································· 14
2.6.柑橘類精油的效用與金桔果皮的利用潛力·················· 14
第3 章材料與方法·········································· 16
3.1 實驗儀器設備與藥品··································· 16
3.1.1 實驗儀器········································· 16
3.1.2 實驗材料與藥品··································· 16
3.2 實驗方法與步驟······································20
3.2.1 壓榨法·········································· 20
3.2.2 蒸氣蒸餾法······································ 20
3.2.3 減壓蒸氣蒸餾法·································· 20
3.3 金桔精油的生物活性測試法····························· 20
3.3.1 金桔精油的抗氧化活性測試法······················ 20
3.3.2 金桔精油的抑菌測試法···························· 21
3.4 金桔精油的分析測試法·································· 22
3.4.1 香氣物質的鑑定方法······························· 22
3.4.2 氣相層析質譜儀(GC-MS)簡介······················· 22
3.4.3 核磁共振儀(NMR)·································· 24
第4 章結果與討論···········································
4.1 金桔精油············································· 34
4.1.1 金桔果皮佔全果實重量百分比······················· 34
4.1.2 三種金桔精油萃取方法的比較與萃取率················ 34
4.1.3 三種金桔精油萃取方法的收集量····················· 35
4.2 金桔精油的DPPH 清除百分率··························· 39
4.3 金桔精油的抑菌測試··································· 39
4.4 金桔精油的測試與主要組成分及主要組成分確認············ 45
4.4.1 金桔精油的GC-MS 與NMR 測試結果················· 45
4.4.2 金桔精油的主要組成分····························· 45
4.4.3 金桔精油的主要組成分確認與應用··················· 45
第5 章結論················································ 51
參考文獻················································· 52
附錄1·····················································55
附錄2·····················································58
附錄3·····················································62
附錄4·····················································67
附錄5·····················································70
附錄6·····················································73
附錄7·····················································82
作者簡介··················································83
圖1、冷壓式柑橘類果皮油製造流程···························· 7
圖2、有代表性的單萜烯類···································· 8
圖3、壓榨法-傳統小型壓榨機流程···························· 25
圖4、壓榨法-工業用大型壓榨機流程·························· 26
圖5、減壓蒸氣蒸餾法流程··································· 26
圖6、DPPH 清除百分率測試法流程··························· 27
圖7、凍乾狀態菌株之活化及保存流程························· 28
圖8、冷凍狀態菌株之活化流程······························· 29
圖9、金桔精油的抑菌法試驗流程····························· 30
圖10、金桔精油以壓榨法-傳統小型壓榨機壓榨收集量············ 38
圖11、金桔精油以蒸氣蒸餾法的萃取收集量····················· 38
圖12、金桔精油以減壓蒸氣蒸餾法的萃取收集量················· 39
圖13、金桔精油GC 層析圖··································· 47
圖14、金桔精油MS 質譜圖··································· 48
圖15、金桔精油1H NMR····································· 49
圖16、金桔精油13C NMR····································· 50
表1、金桔營養成分·········································· 3
表2、萜類的分類············································ 9
表3、精油的一般組成成分··································· 10
表4、一般的氧自由基······································· 12
表5、具有氧化損傷的活性氧································· 13
表6、大腸桿菌培養基M-Endo Agar ···························· 17
表7、綠膿桿菌培養基M-PA-C Agar···························· 18
表8、糞便腸球菌培養基K F Agar ····························· 19
表9、大腸桿菌生長菌落數··································· 31
表10、綠膿桿菌生長菌落數··································· 32
表11、糞便腸球菌生長菌落數································· 33
表12、果皮佔全果實重量百分率······························· 36
表13、金桔精油萃取三種方法的萃取率························· 37
表14、金桔精油的DPPH 清除百分率測試結果·················· 41
表15、金桔精油的大腸桿菌抑菌測試結果······················· 42
表16、金桔精油的綠膿桿菌抑菌測試結果······················· 43
表17、金桔精油的糞便腸球菌抑菌測試結果····················· 44
附圖1、金桔花············································ 55
附圖2、金桔果樹···········································56
附圖3、成熟金桔果實果樹···································· 57
附圖4、金桔-酸味··········································58
附圖5、金桔-甜味··········································59
附圖6、金桔棗-甜味呈橢圓形································· 60
附圖7、實驗金桔的橫切面圖·································· 61
附圖8、金桔果實洗淨擦乾···································· 62
附圖9、金桔果實每粒秤重與記錄······························ 63
附圖10、金桔果實分果皮與果肉······························· 64
附圖11、金桔果皮細分約為1cm2 ······························ 65
附圖12、金桔果皮細分約1cm2 後，果肉可食用·················· 66
附圖13、金桔的果皮需秤重與紀錄····························· 67
附圖14、金桔果皮需細分約為1cm2 ···························· 68
附圖15、實際減壓蒸氣蒸餾過程······························· 69
附圖16、實驗編號A.大腸桿菌培養生長情形····················· 70
附圖17、實驗編號B.綠膿桿菌培養生長情形····················· 71
附圖18、實驗編號C.糞便腸球菌培養生長情形··················· 72
附圖19、實驗編號A.大腸桿菌於金桔精油濃度1.0％形成抑菌環··· 73
附圖20、實驗編號A.大腸桿菌於金桔精油濃度1.5％形成抑菌環··· 74
附圖21、實驗編號A.大腸桿菌於金桔精油濃度2.0％形成抑菌環··· 75
附圖22、實驗編號B.綠膿桿菌於金桔精油濃度1.0％形成抑菌環··· 76
附圖23、實驗編號B.綠膿桿菌於金桔精油濃度1.5％形成抑菌環··· 77
附圖24、實驗編號B.綠膿桿菌於金桔精油濃度2.0％形成抑菌環··· 78
附圖25、實驗編號C.糞便腸球菌於金桔精油濃度1.0％形成抑菌環· 79
附圖26、實驗編號C.糞便腸球菌於金桔精油濃度1.5％形成抑菌環· 80
附圖27、實驗編號C.糞便腸球菌於金桔精油濃度2.0％形成抑菌環· 81
附圖28、試驗三種萃取法所得的金桔精油······················· 82

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