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| (以作者名查詢海洋大學圖書館館藏系統)
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| (以作者名查詢全國博碩士系統)
論文名稱: | 大豆屬一條根抑制 15-LOX成分之鑑定
| Identification of Anti 15-LOX Compounds in I-Tiao Gung (Glycine tomentella Hayata)
研究生: | 王懷德 Huai-Der Wang
指導教授: | 孫寶年 Bonnie Sun Pan, Ph. D.
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: | M95320068
畢業學年度: | 96
語文別: | 中文
關鍵字: | 一條根 I-Tiao Gung
| 15-脂氧合酶 15-lipoxygenase
全文點閱次數: | 16

全文影像連結:

論文全文(pdf)

論文頁數: | 89
摘要: | 摘要
| 大豆屬一條根 (Glycine tomentella Hayata) 具有抑制不飽和脂肪酸自氧化、清除自由基、螯合鐵離子之能力及延緩低密度脂蛋白氧化。於吳郭魚低溫緊迫試驗中，可降低吳郭魚血液流動之起始應力、黏稠度及血小板 5-, 12-, 15- 脂氧合酶活性。在一條根 95% 乙醇萃取物 (GTE) 中，確定含有 daidzein、daidzin、malonyldaidzin、genistein 及 glycitin，但尚有較強於異黃酮類之抗氧化功效成分未被鑑定。本研究為找出 GTE 抗氧化力最高之成分，以抑制 15-脂氧合酶之活性能力為篩選依據，並檢測其對環氧合酶之抑制功效。
| GTE 經乙酸乙酯萃取、silica gel 管柱層析後可得 5 個區分，以第三區分 (GTE-EA-III) 抑制 15-脂氧合酶之能力最高，於濃度 2 µg/mL 之抑制率約為 92.76%。GTE-EA-III 經 silica gel 管柱層析得 GTE-EA-III-A~J 共十區分物，其中 GTE-EA-III-C於濃度 2 µg/mL 對 15-LOX 抑制率約為 98.70%，高於 genistein 且與脂氧合酶抑制劑 (nordihydroguaiaretic acid, NDGA) 於濃度為0.2 µg/mL 之抑制率相似。GTE-EA-III-C 經 silica gel 管柱層析得 GTE-EA-III-C-a~m，共十三區分，經檢測抑制 15-脂氧合酶活性，得知以 GTE-EA-III-C-f & g 區分抗氧化力較強，而其中以 GTE-EA-III-C-g 區分回收量較 f 區分高，因此進一步以半製備型 RP-HPLC分離 GTE-EA-III-C-g ，共分 GTE-EA-III-C-g-1~4 四區分物，GTE-EA-III-C-g-3 抑制 15-脂氧合酶活性之 IC50  為 0.3 µg/mL，劑量為 NDGA 的 7.9倍。
| GTE-EA-III-C-g-3的紫外光光譜中於 245 nm 與 345 nm 有吸收波峰，在 306 nm 有小肩峰；再以 LC/MS 分析，發現陽離子質量為 464 Da，以核磁共振光譜分析，初步推測為 prenylate alkaloids。
|
論文目次: | 目錄
| 摘要    I
| Abstract    III
| 目錄    IV
| 圖目錄    VIII
| 表目錄    X
| 壹、前言    1
| 貳、文獻回顧    4
| 一、豆科植物之異黃酮 (isoflavone) 成分    4
| 二、一條根生理活性    5
| 1. 一條根之來源    5
| 2. 一條根的藥理作用    5
| 3. 一條根之成份及藥理活性    6
| 4. 闊葉大豆之毒性試驗    7
| 三、黃酮類與異黃酮化合物之結構    8
| 四、黃酮類對心血管之影響    10
| 1. 預防紅血球溶血    11
| 2. 預防心血管疾病    13
| 五、異黃酮類之生理活性    14
| 1. 減緩更年期症候群    15
| 2. 預防骨質疏鬆症    16
| 3. 預防癌症    16
| 4. 預防心血管疾病    17
| 六、黃酮類之抗氧化機制    18
| 1. 清除活性氧自由基    18
| 2. 螯合金屬離子    19
| 3. 抑制脂質氧化    20
| 七、發炎及過敏的發生機制    20
| 八、Lipoxygenase (LOX) 生成物及其生理活性    22
| 1. 5-Lipoxygenase (5-LOX) 之氧化產物及發炎反應    23
| 2. 12-Lipoxygenase (12-LOX) 氧化作用及相關疾病(Ref)    23
| 3. 15-Lipoxygenase (15-LOX) 氧化作用及對血管的影響    24
| 九、Cyclooxygenase (COX) 區分及氧化產物    25
| 十、黃酮類抑制 LOX 及 COX 之作用    26
| 参、 實驗流程    27
| 一、 一條根    28
| 二、 方法    28
| 1. 一條根萃取物 (GTE) 之製備    28
| 2. 一條根萃取物之分離    29
| 2.1 乙酸乙酯極性區分    29
| 2.2 silica gel 管柱層析    29
| 2.3 薄層層析分析    30
| 2.4 分離物之 HPLC 分析    31
| 2.5 半製備型HPLC分析    32
| 3. 抗氧化性分析測定脂氧合酶總活性測定    32
| 4. LC/MS/MS測定分析    33
| 5. 核磁共振光譜    33
| 6. 統計分析    33
| 伍、結果與討論    34
| 一、一條根抗氧化成分之分離    34
| 1. 一條根經 95% 乙醇萃取    34
| 2. GTE經乙酸乙酯極性劃分    35
| 3. Silica gel 60 管柱層析    37
| 3.1 GTE-EA 之區分產率及對 15-LOX 抑制活性    37
| 3.2 GTE-EA-III 之區分產率及對 15-LOX 抑制活性    40
| 3.3 GTE-EA-III-C 之區分產率及對 15-LOX 抑制活性    43
| 4. GTE-EA-III-C-g半製備型 HPLC 之區分及對 15-LOX 抑制活性    46
| 二、    一條根區分物之鑑定    49
| 1. 異黃酮之 HPLC 圖譜檢測    49
| 1.1 GTE 及 GTE-EA 之RP-HPLC 圖譜比較    49
| 1.2 GTE-EA-I~V 區分之 RP –HPLC 圖譜    51
| 1.3 GTE-EA-III-A~J 區分之 RP-HPLC 圖譜    53
| 1.4 genistein之 LC/MS 圖譜與定量    59
| 1.5 GTE-EA-III-C-a~m 區分之 RP-HPLC 圖譜    65
| 2. 類黃酮分類之鑑定    70
| 陸、結論    82
| 柒、參考文獻    83
|
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