本研究主要針對綠莖九層塔(Ocimum basilicum L.)、紅莖九層塔(Ocimum basilicum L.)兩種樣品之揮發性抗菌活性成分加以探討。試驗中利用水蒸氣蒸餾法萃取其揮發性成分，並以GC和GC/MS分離及鑑定，再進行抑菌活性分析。
結果顯示，可由綠莖九層塔、紅莖九層塔兩種羅勒之全株、花及葉子三個部位，共鑑定出五十三種化學成分。主要揮發性成分為香艾菊腦(estragole)、沈香醇(linalool)、1,8-桉油酚(1,8-cineole)及甲基丁香酚(methyl eugenol)等。
進一步擇取綠莖、紅莖九層塔精油及其中主要成份香艾菊腦(estragole)進行抗菌活性探討。將三中樣本以濾紙擴散法(Paper Disk Diffusion Technique) 分別就六株細菌進行測試。結果顯示，香艾菊腦對六種菌株均有顯著的抑制效果。而兩種羅勒精油對上皮葡萄球菌（Staphylococcus epidermidis）、金黃色葡萄球菌（Staphylococcus aureus）、沙門氏桿菌（Salmonella typhimurium）及大腸桿菌（Escherichia coli）四株也具有抑制效果。並且發現香艾菊腦及羅勒精油在低於10 ％濃度時對實驗菌株即有抑制效果。

The aim of this study is to identify the volatiles and their antimicrobial activites of Green stem basil (Ocimum basilicum L.), and Red stem basil (Ocimum basilicum L.). Volatiles were extracted through steam distillation, and then characterized by gas chromatography equipped with mass detector before antimicrobial analysis.
Results showed that, more than fifty-three volatiles were identified from overall, fowers, and leaves of two varieties in basil. Mainly estragole, linalool, 1,8-cineole, and methyl eugenol were identified from basil. The analysis further showed that estragole, linalool, 1,8-cineole, and methyl eugenol were the main volatile chemical components.
Green stem basil essential oil, red stem basil essential oil, and the main element ‘estragole’ were then extracted and analyzed for its antimicrobial activities. Paper Disk Diffusion Technique was used to test six species of bacteria separately. The results showed that estragole was able to inhibit all the six species of bacteria.The two essential oils of basils were able to inhibit Staphylococcus epidermidis, Staphylococcus aureus, Salmonella typhimurium, and Escherichia coli. The findings showed that estragole and essential oils at less 10% concentration were inhibitory effects on bacteria.

摘要……….…………………………………………………………………Ⅰ
Abstract………………………………………………………………………Ⅱ
誌謝……………………………………………………………………………Ⅳ
目錄……………………………………………………………………………Ⅴ
圖目錄…………………………………………………………………………Ⅷ
表目錄…………………………………………………………………………Ⅸ
第1章 前言……………………………………………………………………1
第2章 文獻回顧………………………………………………………………3
2.1 羅勒簡介…………………………………………………………………3
2.2 羅勒的植株特色與生長習性....................................................................4
2.3 植物的揮發性成分組成.............................................................................5
2.3.1 植物香氣化合物..................................................................................5
2.3.2 植物精油之萃取法............................................................................10
2.3.2.1 冷壓榨法......................................................................................10
2.3.2.2 水蒸餾法......................................................................................10
2.3.2.3 水蒸氣蒸餾法..............................................................................12
2.3.2.4溶劑萃取法...................................................................................12
2.3.2.5 油脂分離法..................................................................................12
2.3.2.6 分餾法..........................................................................................12
2.3.2.7 同時水蒸氣蒸餾-有機溶劑抽取法............................................12
2.3.2.8 冷吸法..........................................................................................13
2.3.2.9 超臨界流體香精抽取法..............................................................13
2.3.2.10 微波抽取法...............................................................................13
2.4 羅勒應用..................................................................................................14
2.5 精油抗菌活性..........................................................................................15
2.5.1 香料的殺菌力 ..................................................................................15
2.5.2 精油抗菌活性....................................................................................15
2.5.3 萜類化合物的抗菌活性....................................................................16
2.5.3.1 單萜類化合物的抗菌活性..........................................................16
2.5.3.2 倍半萜類化合物的抗菌活性......................................................18
2.5.4 抗菌活性試驗方法............................................................................18
2.5.4.1 洋菜稀釋法..................................................................................18
2.5.4.2 濾片擴散法..................................................................................19
第3章 材料與方法...........................................................................................20
3.1 實驗材料...................................................................................................20
3.1.1 羅勒....................................................................................................20
3.1.2 藥品試劑............................................................................................20
3.1.2.1 標準品..........................................................................................20
3.1.2.2 藥品..............................................................................................20
3.1.3 儀器設備............................................................................................20
3.2 實驗方法...................................................................................................22
3.2.1 實驗架構............................................................................................22
3.2.2 羅勒之栽培........................................................................................22
3.2.3 精油之製備........................................................................................22
3.2.4 氣相層析儀及氣相層析質譜儀分析................................................24
3.2.4.1 氣相層析儀..................................................................................24
3.2.4.2 氣相層析質譜儀分析..................................................................24
3.2.5 抗菌性活性測定................................................................................25
3.2.5.1 試驗菌株......................................................................................25
3.2.5.2 培養基組成份..............................................................................25
3.2.5.3 菌種保存與更新..........................................................................27
3.2.5.4 試驗菌種之製備..........................................................................27
3.2.5.5 紙錠擴散法之測定......................................................................27
3.2.6 統計分析............................................................................................28
第4章 結果與討論...........................................................................................29
4.1 羅勒精油收率..........................................................................................29
4.2 羅勒精油揮發性成分分析與質譜鑑定...................................................29
4.3 羅勒精油與香艾菊腦抑菌活性之探討...................................................56
第5章 結論.......................................................................................................66
參考文獻.............................................................................................................68
作者簡介.............................................................................................................75
圖 目 錄
頁次
圖1、異戊二烯結構式…………………………………………………………8
圖2、植物性香料的採油法……………………………...……………………11
圖3、精油中具有抗菌活性成分之化合物………………..…………………17
圖4、簡易精油萃取裝置圖…………………………………..………………21
圖5、實驗流程設計圖…………………………………………..……………23
圖6、羅勒採樣的四個不同部位：嫩葉、老葉、花、全株…..……………31
圖7、綠莖九層塔全株揮發性成分之氣相層析TIC圖…………..…………33
圖8、綠莖九層塔花揮發性成分之氣相層析TIC圖…………………..……34
圖9、綠莖九層塔葉子揮發性成分之氣相層析TIC圖……………………35
圖10、紅莖九層塔全株揮發性成分之氣相層析TIC圖……………………36
圖11、紅莖九層塔花揮發性成分之氣相層析TIC圖………………………37
圖12、紅莖九層塔葉子揮發性成分之氣相層析TIC圖……………………38
圖13、羅勒不同部位精油含量之比較……………………………...………46
圖14、羅勒中香艾菊腦的質譜 ……………………………………..………47
圖15、羅勒中沈香醇的質譜 …………………………………………..……48
圖16、羅勒中1,8-桉油酚的質譜 …………………………………..………49
圖17、羅勒中甲基丁香酚的質譜 …………………………………..………50
圖18、羅勒中樟腦的質譜 ……………………………………………..……51
圖19、羅勒中β-欖香烯的質譜 …………………………………..…………52
圖20、羅勒中α-佛手柑油烯的質譜 ………………………………..………53
圖21、羅勒中大根香葉烯-D的質譜 ………………………………….……54
圖22、羅勒中α-杜松烷醇的質譜 ……………………………………….…55
圖23、不同精油對Staphylococcus epidermidis菌株抑制圈之影響 …...…60
圖24、不同精油對Listeria monocytogenes菌株抑制圈之影響 ……….…61
圖25、不同精油對Staphylococcus aureus菌株抑制圈之影響 ……………62
圖26、不同精油對Pseudomonas aeruginosa菌株抑制圈之影響 ……...…63
圖27、不同精油對Salmonella typhimurium菌株抑制圈之影響 …….……64
圖28、不同精油對Escherichia coli菌株抑制圈之影響 ………………..…65

表 目 錄
頁次
表1、不同羅勒品種園藝性狀比較...................................................................6
表2、不同羅勒品種植株生育比較.................................................................7
表3、萜的分類.................................................................................................9
表4、試驗菌株及其來源................................................................................26
表5、不同覆蓋遮光率之羅勒生長情形.........................................................30
表6、兩種不同品種的羅勒精油之萃取率....................................................32
表7、綠莖羅勒及紅莖羅勒中全株、花、葉三部分揮發性成分
組成的GC-MS波峰面積比較……………………………………..…41
表8、綠莖九層塔精油對試驗菌株抑制圈之影響 …………………………57
表9、紅莖九層塔精油對試驗菌株抑制圈之影響 …………….……..……58
表10、香艾菊腦對試驗菌株抑制圈之影響 ………………….……………59

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