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研究生:曾家澤
研究生(外文):Chia-Tse Tseng
論文名稱:蘿蔔嬰、綠花椰菜芽及油菜芽甲醇萃取物之抗氧化活性研究
論文名稱(外文):Study on the antioxidant activity of methanolic extracts of radish sprouts, broccoli sprouts and rapeseed sprouts
指導教授:翁瑞光 林世斌
指導教授(外文):Ray-Guang Wong, Shih-Bin Lin
學位類別:碩士
校院名稱:國立宜蘭大學
系所名稱:食品科學系碩士班
學門:農業科學學門
學類:食品科學類
論文種類:學術論文
論文出版年:2008
畢業學年度:96
語文別:中文
論文頁數:88
中文關鍵詞:芽菜抗氧化十字花科一氧化氮巨噬細胞RAW264.7
外文關鍵詞:sproutsantioxidantcruciferous vegetablesnitric oxideRAW264.7 macrophage
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本研究旨在探討不同發芽時間(39天)對蘿蔔嬰(radish sprouts)、綠花椰菜芽(broccoli sprouts)及油菜芽(rapeseed sprouts)甲醇萃取物之抗氧化活性及有效成分之影響,藉此篩選出最佳的抗氧化活性天數,以進行體外細胞抗發炎實驗,了解抗氧化活性與抑制RAW264.7細胞產生NO之相關性。
抗氧化實驗的結果顯示,蘿蔔嬰、綠花椰菜芽及油菜芽於不同發芽時間皆有抑制亞麻油酸氧化、還原力、清除DPPH自由基及螯合亞鐵離子之能力,其中皆以發芽第七天有較佳的抗氧化活性。在抑制亞麻油酸氧化之效果其IC50分別為33 g/mL、0.5 g/mL及40.5 g/mL。蘿蔔嬰、綠花椰菜芽及油菜芽發芽時所含之總多酚類化合物及總葉綠素含量會隨發芽時間有不同的變化,但皆以發芽第七天的含量較高,其中總酚類化合物與抑制亞麻油酸氧化、還原力及清除DPPH自由基皆有顯著之相關 (p < 0.05),其相關係數分別為0.792、0.900及0.699。體外細胞抗發炎實驗的結果顯示,發芽第7天的芽菜甲醇萃取物濃度在83 g/mL以下對細胞沒有毒性,於此濃度下蘿蔔嬰、綠花椰菜芽能抑制LPS誘導RAW264.7巨噬細胞生成的NO,其抑制率各為20.5%及36.3%,而油菜芽則沒有抑制NO生成的效果,此外芽菜甲醇萃取物在抑制體外細胞NO生成與抑制亞麻油酸氧化、還原力、清除DPPH自由基及螯合亞鐵離子皆呈正相關(r = 0.682、0.715、0.683及0.593),顯示抗氧化力表現越高,在抑制巨噬細胞生成NO的能力也越強。
Effects of germination time on the production of antioxidant activity and antioxidant compounds of methanolic extracts of radish sprouts, broccoli sprouts and rapeseed sprouts were studied. The correlation between optimal antioxidation and antiinflammatory activity on RAW264.7 macrophage cell was also investigated.
The results showed that the methanolic extracts of three kinds of sprouts from different germination time had ability of inhibition on the linoleic acid oxidation, reducing power, scavenging DPPH free radicals and Fe2+chelating activity. The seventh-day grown sprouts had the best antioxidant activity. The IC50 of linoleic acid oxidation by radish sprouts, broccoli sprouts and rapeseed sprouts showed 33 g/mL, 0.5 g/mL and 40.5 g/mL, respectively. The contents of total phenolic compounds and total chlorophyll contents in the seed sprouts were changing during the germination time and the highest contents occurred at the seventh day. There are significant correlation between phenolic compounds and inhibition on the linoleic acid oxidation, reducing power and scavenging DPPH free radicals (r = 0.792, 0.900 and 0.699). In the in vitro of experiment antiinflammatory the results showed that the extracts (83 g/mL) of radish sprouts, broccoli sprouts rapeseed sprouts had no cytotoxic effect on RAW264.7 macrophage. The extracts (83 g/mL) of radish sprouts and broccoli sprouts were able to suppress the NO production by LPS-induced RAW264.7 macrophage (20.5% and 36.3%), but rapeseed sprouts was not. Moreover we found the positive correlation between the suppression of NO production and inhibition on the linoleic acid oxidation, reducing power, scavenging DPPH free radicals and Fe2+chelating activity (r = 0.682, 0.715, 0.683 and 0.593). Experiments data showed that the high antioxidant activity also had good suppressing power on NO produced by RAW264.7 macrophage.
中文摘要……………………………………………………………… I
英文摘要……………………………………………………………… III
目錄…………………………………………………………………… V
表目錄………………………………………………………………… IX
圖目錄………………………………………………………………… X
壹、前言……………………………………………………………… 1
貳、文獻整理………………………………………………………… 5
一、氧化與抗氧化防禦……………………………………………… 5
(一)何謂反應性氧分子及反應性氮分子…………………………… 5
(二)反應性氧分子及反應性氮分子之來源………………………… 5
(三)氧化傷害與疾病………………………………………………… 10
(四)抗氧化防禦物質………………………………………………… 12
二、芽菜之介紹……………………………………………………… 13
(一)發芽與芽菜……………………………………………………… 13
(二)發芽與營養成分的改變………………………………………… 14
(三)芽菜之抗氧化活性……………………………………………… 15
三、十字花科蔬菜之介紹…………………………………………… 17
(一)十字花科蔬菜及其營養價值…………………………………… 17
(二)十字花科蔬菜之抗氧化活性…………………………………… 18
(三)十字花科之生理活性…………………………………………… 19
(四)蘿蔔之簡介……………………………………………………… 21
(五)綠花椰菜之簡介………………………………………………… 21
(六)油菜之簡介……………………………………………………… 22
參、材料與方法……………………………………………………… 23
一、實驗架構………………………………………………………… 23
二、實驗材料………………………………………………………… 23
(一)樣品……………………………………………………………… 23
(二)藥品……………………………………………………………… 24
(三)儀器設備………………………………………………………… 25
三、樣品製備………………………………………………………… 25
(一)芽菜培育………………………………………………………… 25
(二)芽菜樣品之製備………………………………………………… 26
(三)芽菜萃取液之製備……………………………………………… 26
四、實驗方法與流程………………………………………………… 27
(一)抗氧化成分分析測定…………………………………………… 27
1.總多酚類化合物含量測定………………………………………… 27
2.總葉綠素含量測定………………………………………………… 27
(二)抗氧化活性測定………………………………………………… 28
1.硫氰酸鐵法測定…………………………………………………… 28
2.還原力測定………………………………………………………… 28
3.DPPH自由基清除能力測定………………………………………… 28
4.亞鐵離子螯合能力測定…………………………………………… 29
(三)體外細胞實驗…………………………………………………… 29
1.細胞培養…………………………………………………………… 29
2.細胞存活率測定…………………………………………………… 30
3.NO生成量測定…………………………………………………… 30
(四)萃取物對抗氧化實驗之半數清除濃度反應模式……………… 31
(五)統計分析………………………………………………………… 31
肆、結果與討論……………………………………………………… 32
一、蘿蔔嬰、綠花椰菜芽及油菜芽於不同發芽時期之生長變化… 32
二、不同發芽時期之蘿蔔嬰、綠花椰菜芽及油菜芽甲醇萃取物抗氧化成分之變化…………………………………………………………………… 32
(一)總酚類化合物……………………………………………………32
(二)總葉綠素…………………………………………………………34
三、不同發芽時期之蘿蔔嬰、綠花椰菜芽及油菜芽甲醇萃取物之抗氧化性質……………………………………………………………………… 36
(一)總抗氧化力……………………………………………………… 36
(二)還原力…………………………………………………………… 39
(三)DPPH自由基清除力…………………………………………… 41
(四)亞鐵離子螯合能力……………………………………………… 44
四、蘿蔔嬰、綠花椰菜芽及油菜芽萃取物之體外細胞實驗……… 47
(一)細胞存活率……………………………………………………… 47
(二)一氧化氮生成量之測定………………………………………… 49
伍、結論……………………………………………………………… 53
陸、參考文獻………………………………………………………… 54
柒、表………………………………………………………………… 71
捌、圖………………………………………………………………… 75
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