臺灣博碩士論文加值系統

本研究以紅、綠紫蘇(Perilla frutescens) 葉為原料，將其攪碎後，以水蒸氣蒸餾技術萃取精油。所得精油再以矽膠管柱層析以不同溶劑區分之，其區分物再以氣相層析質譜分析儀（GC/MS）鑑定其揮發性成分。同時也以溶劑(水及乙醇)萃取紅、綠紫蘇葉中之機能性成分，並探討紅、綠紫蘇葉精油、水萃物及乙醇萃取物之抗氧化性、以及抗菌及抗發炎能力之研究。研究結果如下:
紅、綠紫蘇葉精油之收率分別為0.04%及0.03%。在紅紫蘇葉精油中共鑑定出40種化合物，其主要的成分為perilla aldehyde、limonene其含量分別為54.35%及23.81%，紅紫蘇葉精油以矽膠管柱層析法區分，其Pentane層的主要成分為trans-caryopnyllene，含量為38.94%，在Ether層的主成分為perilla aldehyde其含量為93.19%。綠紫蘇葉精油中共鑑定出43種化合物，其主成分為perilla aldehyde及limonene，其含量分別為65.26%及12.49%。經矽膠管柱層析法區分後，其Pentane層的主成分為cis,trans-α-farnesene其含量為41.96%，在Ether層的主成分為perilla aldehyde其含量為84.40%。
在抗氧化活性分析中，清除DPPH自由基能力，當濃度為5mg/mL時以紅紫蘇葉水萃取物清除力最佳達64.52%。分析螯合亞鐵離子能力，當濃度為5mg/mL時以綠紫蘇葉水萃取物螯合力最佳達34.87%。清除SOD-like之能力，當濃度為0.5mg/mL時以綠紫蘇Ether區分物清除力最佳達56.52%。清除H2O2之能力，當濃度0.5mg/mL時以紅紫蘇Ether區分物清除力最佳達56.12%。
機能性成份之總酚類化合物(Total polyphenolics)、芸香苷(Rutin)、檞皮素(Quercetin)含量分析結果，每100克中紅、綠紫蘇葉總酚含量分別為578.49、614.02毫克，芸香苷含量分別為20.44及13.92毫克，檞皮素77.41及29.65毫克。
在抗菌活性分析中，未區分前紅、綠紫蘇葉精油在濃度0.1%時對不同病原菌其抑制效果為70%以上，經區分後Pentane層精油其對各病原菌的抑制效果皆不好，而Ether層精油在濃度0.1%時對E. coli、S. aureus、V.parahaemolyticus、S. typhimurium皆可高達95%以上的抑菌效果，對T. mentagrophytes只有85%的抑菌效果，另外也發現綠紫蘇葉精油對H.pylori的抑菌效果比紅紫蘇葉精油還要好。
在抗發炎試驗中，紅、綠紫蘇水萃取物幾乎抑制TNF-α的產生，結果顯示，水萃取物抑制TNF-α產生是優於紅、綠紫蘇乙醇萃取物，而當中又以紅紫蘇比綠紫蘇有更好的抗發炎效果。
綜合上述結果，紫蘇葉確實具有抗氧化、抗菌及抗發炎之能力。
關鍵字：紫蘇、精油、抗氧化、抗菌、抗發炎

Red and green Perilla frutescens leaves were examined for their antioxidant antibacterial, and anti-inflammatory bioactivities. Essential oils (EO) were first separated by the steam distillation. The obtained EO were further subjected to solvent fractionation on a silica gel chromatorgraphy. The separated sub EO fractions were analyzed by GC/MS.
The yields of the essential oil were 0.04% and 0.03% for the red and green leaves of Perilla frutescens, respectively. Fourty kinds of volatiles were found in the essential oil of red Perilla frutescens, among which the major components were perilla aldehyde and limonene, reaching respectively 54.35% and 23.81% in the whole essential oils. On fractionation with pentane, trans-caryopnyllene was found to be the major chemical and reached a content of 38.94%. While the ether layer contained 93.19% of perilla aldehyde. Fourty three kinds of volatiles were found in the essential oil of green Perilla frutescens, perilla aldehyde and limonene, the major two constituents, were found to reach 65.26 and 12.49%, respectively. The corresponding pentane layer contained 41.96% of trans-caryopnyllene, and the ether layer 84.40% of perilla aldehyde.
By antioxidant analysis, the aqueous extract of red leaves at concentration of 5mg/mL showed 64.52% of DPPH free radical scavenging capability.In contrast, the aqueous extract of green leaves at the same concentration exhibited a 34.87% of Fe2+-chelating capability. Its ether fractioante at 0.5 mg/mL showed a 56.52% of SOD scavenging capability. In contrast, the ether extract of red leaves exhibited a 56.12% H2O2 cleavage capability.
The total polyphenolics were analyzed. Data showed that each 100g of the red/green leaves contained a total poplyphenolics of 578.49/614.02mg. The content of rutin and quercetin reached 20.44/13.92mg and
77.41/29.65mgrespectively.
By antibacterial analysis, we recognized that the crude essential oil at 0.1% revealed 70% of antibacterial properties. On a chromatographic separation, the ether fractionate attained a 95% of antibacterial activity against the common pathogens E. coli, S. aureus, V.parahaemolyticus, and S. typhimurium, while only 85% effective against the strain T. mentagrophytes, comparing to the better effect of the essential oil of green leaves in inhibition of H. pylori.
By anti-inflammatory analysis, we showed that all the aqueous extracts of red and green leaves revealed substantial effect to suppress the expression of TNF-α, and indeed more potent than the ethanol extracts. In contrast, the extracts of red leaves possessed better anti-inflammatory bioactivity.
In conclusion, Perilla frutescens leaves, despite the red or green, have the merit to provide moderate anti-oxidants, anti-bacterial and anti-inflammatory capability.

中文摘要………………………………………………………… 1
英文摘要………………………………………………………… 3
前言……………………………………………………………… 5
文獻回顧………………………………………………………… 7
一、紫蘇簡介…………………………………………………… 7
(ㄧ)種類及特性………………………………………………… 7
(二)品種………………………………………………………… 7
(三)紫蘇之生理活性及機能性分…………………………… 9
(四) 精油特性……………………………………………… 16
(五) 揮發性成分…………………………………………… 17
(六) 精油萃取方式………………………………………… 20
(七) 精油分離與鑑定……………………………………… 22
(八)精油抗菌活性之研究…………………………………… 23
(九) 巨噬細胞與抗發炎的關係…………………………… 26
實驗架構………………………………………………………… 29
材料與方法……………………………………………………… 32
結果與討論……………………………………………………… 55
ㄧ、 紅、綠紫蘇葉成分及機能性成分分析…………………… 55
(ㄧ) ㄧ般組成份含量之變化……………………………… 55
(二) 紅、綠紫蘇葉水及乙醇萃取物有機酸之含量………… 55
(三) 紅、綠紫蘇葉水及乙醇萃取物總酚、芸香苷及槲
皮酮之含量……………………………………………… 56
(四) 紅、綠紫蘇葉水及乙醇萃取物葉黃素之含量………… 57
二、紅、綠紫蘇精油香氣成分之探討…………………………… 58
三、紅、綠紫蘇水及乙醇萃取物之抗氧化性質………………… 60
(ㄧ) 紅、綠紫蘇葉水及乙醇萃取物之收率………………… 60
(二) 紅、綠紫蘇葉水及乙醇萃取物、精油及區分物清
除DPPH自由基之能力………………………………… 60

(三) 紅、綠紫蘇葉水及乙醇萃取物、精油及區分物螯
合亞鐵離子之能力…………………………………… 62
(四) 紅、綠紫蘇葉精油及其區分物清除超氧陰離子之能力 63
(五) 紅、綠紫蘇葉精油及其區分物清除過氧化氫之能力 64
四、紅、綠紫蘇葉精油及其區分物之抑菌活性評估………… 64
(一) 紅、綠紫蘇葉精油及其區分物對大腸桿菌之抑菌效果 66
(二) 紅、綠紫蘇葉精油及其區分物對金黃色葡萄球菌之抑
菌效果………………………………………………… 66
(三) 紅、綠紫蘇葉精油及其區分物對腸炎弧菌之抑菌效果 67
(四) 紅、綠紫蘇葉精油及其區分物對沙門氏菌之抑菌效果 67
(五) 紅、綠紫蘇葉精油及其區分物對足癬菌之抑菌效果… 68
五、紅、綠紫蘇葉水及乙醇萃取物之抗發炎活性評估………… 69
結論………………………………………………………………… 117
參考文獻…………………………………………………………… 119









圖次
圖一 紫花紅葉紫蘇…………………………………………… 9
圖二 白花青葉紫蘇…………………………………………… 9
圖三 芸香苷的化學結構……………………………………… 14
圖四 槲皮素的化學結構……………………………………… 15
圖五 L P S 誘發巨噬細胞T N F - α表現p a t h w a y……… 28
圖六 紅、綠紫蘇葉水及乙醇萃取物清除DPPH自由基
之能力…………………………………………………… 84
圖七 紅紫蘇葉精油及其區分物清除DPPH之能力………… 86
圖八 綠紫蘇葉精油及其區分物清除DPPH之能力………… 88
圖九 紅、綠紫蘇葉水及乙醇萃取物對亞鐵離子之螯合能力… 90
圖十 紅紫蘇葉精油及其區分物對亞鐵離子之螯合能力…… 92
圖十一 綠紫蘇葉精油及其區分物對亞鐵離子之螯合能力…… 94
圖十二 紅紫蘇葉精油及其區分物對SOD-like之清除能力…… 96
圖十三 綠紫蘇葉精油及其區分物對SOD-like之清除能力…… 98
圖十四 紅紫蘇葉精油及其區分物對H2O2之清除能力……… 100
圖十五 綠紫蘇葉精油及其區分物對H2O2之清除能力……… 102
圖十六 水蒸氣蒸餾所得紅、綠紫蘇葉精油及其區分物對
大腸桿菌之抑菌率……………………………………
104
圖十七 水蒸氣蒸餾所得紅、綠紫蘇葉精油及其區分物對
金黃色葡萄球菌之抑菌率……………………………
106
圖十八 水蒸氣蒸餾所得紅、綠紫蘇葉精油及其區分物對
腸炎弧菌之抑菌率……………………………………
108
圖十九 水蒸氣蒸餾所得紅、綠紫蘇葉精油及其區分物對
沙門氏菌之抑菌率…………………………………… 110
圖二十 水蒸氣蒸餾所得紅、綠紫蘇葉精油及其區分物對
足癬菌之抑菌率……………………………………… 112
圖二十一 紅、綠紫蘇葉水萃取物影響RAW264.7巨噬
細胞 TNF-α的產生…………………………………… 113
圖二十二 紅、綠紫蘇葉乙醇萃取物影響RAW264.7巨噬
細胞 TNF-α的產生…………………………………… 114
圖二十三 紅、綠紫蘇葉水萃取物對巨噬細胞 RAW 264.7細
胞存活率之影響……………………………………… 115
圖二十四 紅、綠紫蘇葉乙醇萃取物對巨噬細胞 RAW
264.7細胞存活率之影響……………………………… 116










表次
表一 紫蘇中芸香苷與槲皮素分析之HPLC沖提梯度……… 37
表二 紅、綠紫蘇葉之ㄧ般組成分分析………………………… 72
表三 紅、綠紫蘇葉水及乙醇萃取物中有機酸之含量………… 73
表四 紅、綠紫蘇葉水及乙醇萃取物中總酚、芸香苷、槲皮
素、葉黃素之含量……………………………………… 74
表五 以水蒸氣蒸餾所得紅紫蘇葉精油之揮發性化合物百
分組成之比較…………………………………………… 75
表六 以水蒸氣蒸餾所得綠紫蘇葉精油之揮發性化合物百
分組成之比較…………………………………………… 78
表七 比較紅、綠紫蘇葉精油中不同揮發性化合物之組成 81
表八 紅、綠紫蘇葉水及乙醇萃取物之收率………………… 82
表九 紅、綠紫蘇葉精油及其區分物之收率………………… 82
表十 紅、綠紫蘇葉水及乙醇萃取物清除DPPH之能力…… 83
表十一 紅紫蘇葉精油及其區分物清除DPPH之能力………… 85
表十二 綠紫蘇葉精油及其區分物清除DPPH之能力………… 87
表十三 紅、綠紫蘇葉水及乙醇萃取物對亞鐵離子之螯合能力 89
表十四 紅紫蘇葉精油及其區分物對亞鐵離子之螯合能力…… 91
表十五 綠紫蘇葉精油及其區分物對亞鐵離子之螯合能力…… 93
表十六 紅紫蘇葉精油及其區分物對SOD-like之清除能力…… 95
表十七 綠紫蘇葉精油及其區分物對SOD-like之清除能力…… 97
表十八 紅紫蘇葉精油及其區分物對H2O2之清除能力………… 99
表十九 綠紫蘇葉精油及其區分物對H2O2之清除能力………… 101
表二十 水蒸氣蒸餾所得紅、綠紫蘇葉精油及其區分物對大腸
桿菌之抑菌率…………………………………………… 103
表二十一 水蒸氣蒸餾所得紅、綠紫蘇葉精油及其區分物對金黃
色葡萄球菌之抑菌率…………………………………… 105
表二十二 水蒸氣蒸餾所得紅、綠紫蘇葉精油及其區分物對腸炎
弧菌之抑菌率………………………………………… 107
表二十三 水蒸氣蒸餾所得紅、綠紫蘇葉精油及其區分物對沙門
氏菌之抑菌率………………………………………… 109
表二十四 水蒸氣蒸餾所得紅、綠紫蘇葉精油及其區分物對足癬
菌之抑菌率…………………………………………… 111

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