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論文名稱: | 大豆屬一條根異黃酮類化合物含量與抗氧化性之鑑定
| Identification and antioxidant activity of isoflavones in I- Tiao Gung extract of Glycine tomentella Hayata
研究生: | 陳冠良 Kuan-Ling Chen
指導教授: | 孫寶年 Bonnie Sun
| 廖若川 York Liao
學位類別: | 碩士
學校名稱: | 國立臺灣海洋大學
系所名稱: | 食品科學系
學號: | M91320018
畢業學年度: | 93
語文別: | 中文
關鍵字: | 大豆屬一條根 Glycine tomentella Hayata
| 抗氧化 antioxidation
全文點閱次數: | 8

全文影像連結:

論文全文(pdf)

論文頁數: | 91
摘要: | 金門大豆屬(Glycine tomentella Hayata)一條根宣稱有舒筋活血、鎮痛解熱、解毒消腫等功能。本實驗室已知大豆屬一條根中含黃酮類與異黃酮類，具抑制脂氧合酶 (lipoxygenase, LOX)活性及抗氧化能力，故本研究為比較大豆屬一條根以acetonitrile與methanol加鹽酸水解後，異黃酮類化合物含量之變化與對抗氧化性及脂氧合酶活性之影響。
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| 大豆屬一條根95％乙醇萃取物 (95E) 以acetontrile與鹽酸混合，其萃取層之262 nm吸光值會隨0~0.3 N鹽酸濃度之提高而上升 (r2= 0.95)，水解物中daidzein含量隨著鹽酸濃度的提升而增加 (r2= 0.98)，以methanol與鹽酸混合，其水解層之262 nm吸光值會隨0.2與2 N鹽酸濃度及1與2小時之鹽酸水解時間提高而上升 (r2= 0.98)，水解物中daidzein含量隨著鹽酸濃度的提升而增加 (r2= 0.99)，此鹽酸水解物經HPLC分析含daidzin, glycitin, malonyldaidzin, daidzein, glycitein及 genistein，其中daidzin之含量隨著鹽酸濃度提升而減少，daidzein之含量隨之增加，表示daidzin會被鹽酸水解醣基而形成daidzein。
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| 95E以acetonitrile加0~0.3 N鹽酸水解物與以methanol加0.2 N及2 N鹽酸水解1及2小時水解物之掃除DPPH自由基試驗、對亞麻油酸(linoleic acid)自氧化及黃豆脂氧合酶 (soybean lipoxygenase, LOX) 抑制作用之半抑制濃度 (IC50)，以methanol加2 N鹽酸水解2小時之水解層 (MeOH-2N-2h)抑制效果最高，IC50值分別為0.046, 0.13及0.01 mg/mL，加鹽酸是爲了將95E中含醣苷之異黃酮水解成不含醣苷之異黃酮，以提升抗氧化與抑制脂氧合酶活性之能力，此三者之抑制作用均隨著水解物中daidzein及genistein含量之增加而提升，故鹽酸水解物對掃除DPPH自由基、抑制亞麻油酸自氧化及脂氧合酶之抑制作用隨daidzein及genistein含量增加而提升。
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| 95E鹽酸水解物以methanol為酸水解溶劑萃取出之daidzein 含量較以acetonitrile為溶劑萃取可增加13％，其中以MeOH-2N-2h之抗氧化性較高，將此水解物以製備型TLC (silica gel)分離成四個區分，再將第四區分以製備型TLC (silica gel)再分離成四個區分，以分析型HPLC、TLC及呈色反應，可知其為flavonol、flavanone、isoflavones及chalcone類化合物，為抗氧化力及抑制脂氧合酶能力的提升原因。
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論文目次: | 中文摘要………………………………………………………………Ι
| 英文摘要………………………………………………………………Ⅲ
|
| 壹、前言    …………………..……………………………..1
| 貳、文獻整理    ……………………………………………..3
| 一、    黃酮類與異黃酮類化合物之結構…………………….....3
| 二、    黃酮類之生理功能…...……………...………………….5
| 1.    預防紅血球溶血………………………………………………….5
| 2.    預防心血管疾病………………………………………………….5
| 三、    異黃酮之生理功能………………………………………….7
| 1.    生理功能………………………………………………………….7
| 2.    抑制血小板活化………………………………………………….9
| 3.    活性與非活性異黃酮之差異……………………………………..9
| 四、脂氧合酶對人體所造成的影響….………………………9
| 1. 與動脈粥狀硬化之關係……………..………………..…..10
| 2. 與血液黏度之關係……………………………………...11
| 3. 脂氧合酶與血小板之關係……………………………....12
| 五、異黃酮之抗氧化性……………….……………...………13
| 1.    螯合金屬離子………………………………………...13
| 2.    清除活性氧或自由基………………………………..14
| 六、異黃酮之分析方法………………………………………14
| 參、材料與方法…………………………….………………16
| 一、材料    …………………………………………………….16
| 二、分析方法    …………………………………………….16
| 實驗流程  …………………………………………….17
| 1.一條根之萃取………………………..………..……......18
| 2.酸水解…………………………………..…….………...18
| (a) 以acetonitrile加鹽酸水解…………………………………18
| (b) 以methanol加鹽酸水解…………………………………...18
| 3.脂氧合酶總活性測定…………………………………..19
| 4.掃除自由基試驗………..………….……..……...……..19
| 5.自氧化之抑制作用…………….………………..……...20
| 6.分離物之HPLC分析…..…..……………………..……21
| 7.製備式HPLC……….…...……………………..…...…..22
| 8.TLC分析法……………..………………………………22
| 9.製備式TLC…………………………………………..…23
| 10.呈色反應………………………..……………..……....24
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| 肆、結果與討論…………………………………..…………25
| 1.大豆屬一條根95％乙醇萃取物之產率…………….25
| 2.以acetonitrile加酸水解……………………………..25
| 3.掃除自由基的能力…………………………………..28
| (a)以acetonitrile加鹽酸水解之掃除自由基能力………28
| (b)以methanol加酸水解之掃除自由基能力……………28
| 4.對亞麻油酸自氧化之抑制作用……………………..30
| (a)以acetonitrile加鹽酸水解之掃除自由基能力………30
| (b)以methanol加酸水解之掃除自由基能力……………30
| 5.對脂氧合酶活性的抑制……………………………..32
| (a)以acetonitrile加鹽酸水解之掃除自由基能力……….32
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| (b)以methanol加酸水解之掃除自由基能力……………32
| 6.以TLC及呈色反應判別化合物種類………………...35
| 7.HPLC之定量、分離與鑑定………………………….39
| 8.大豆屬一條根經鹽酸水解後之活性成分分離……...48
| 9.以製備式HPLC分離黃酮類化合物…………………50
| 10.以紫外燈判定化合物最大吸收波峰及種類………..50
| 11.以NMR鑑定化合物結構…………………………...51
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| 伍、結論……………………………………………………..59
| 陸、參考文獻………………..………………………………61
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