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研究生:洪筱鈞
研究生(外文):Hung Hsiao-Chun
論文名稱:大豆中分離高純度Genistin之程序
論文名稱(外文):The isolation process for Genistin of high purity from soybeans
指導教授:黃銘賢
學位類別:碩士
校院名稱:國立高雄應用科技大學
系所名稱:化學工程系碩士班
學門:工程學門
學類:化學工程學類
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:71
中文關鍵詞:大豆配糖型異黃酮大孔樹脂去糖型異黃酮
外文關鍵詞:soybeansisoflavone glycosidesmacroreticular resinisoflavone aglycones
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由於大豆異黃酮在抗癌、舒緩婦女更年期徵狀和預防酒精中毒等方面都有相當多的報導,因此在製藥、食品及化妝品等領域都將擁有非常廣闊的應用前景。本研究旨在發展一套簡易、快速又高效的萃取分離程序,從大豆全豆中以60%甲醇溶劑萃取,萃取液經濃縮後形成大豆蜜,經由HP-2MG樹脂管柱吸附異黃酮,以60%甲醇脫附製得含去糖型與配糖型之異黃酮產物(daidzein、genistein、daidzin、glycitin與genistin),產率為0.26%;再將所得產物藉由HP-2MG樹脂管柱進行分離,以pH=12氫氧化鈉溶液脫附,得到產率為0.04%,透過FT-IR、UV-Vis與HPLC分析鑑定為高純度的genistin成分;樹脂管柱接續以60%甲醇作完全脫附,得到配糖型大豆異黃酮混合物,產率為0.11%,經由相同儀器檢測,得知該混合物是由genistin、daidzin與glycitin組成。
Soy isoflavones are often reported on the prevention of cancer, women menopausal symptoms and alcoholism, and have broad application in the pharmaceutics, foodstuff and cosmetics. This research purports to develop a simple, fast and effective extraction and separation procedure. The methanol of 60 per cent was used to extract the whole soy beans. The extract was formed the soy molasses after being concentrated and absorbed by the HP-2MG column. The yield after being desorbed by the methanol of 60 per cent was 0.26 per cent of isoflavone aglycones and isoflavone glycosides. Then the products was passed through the HP-2MG column to carry on the separation procedure. The desorption was made by sodium hydroxide of 0.01N. The 0.04% yield was obtained. The FT-IR, UV-Vis and HPLC analysis identified it as a high-grade component of genistin. Subsequently using 60 percent methanol to pass through the same column to complete desorption. The mixture of isoflavone glycosides was finally obtained. The yield was 0.11%. The mixture is composed of genistin, daidzin and glycitin.
致謝 ............................................................Ⅰ
中文摘要 ........................................................ Ⅱ
Abstract ...................................................... Ⅲ
目錄 ........................................................... Ⅳ
圖目錄 ......................................................... Ⅶ
表目錄 ......................................................... Ⅸ
壹、文獻回顧 .................................................... 1
1.1 大豆概述 ................................................ 1
1.2 大豆蛋白(Soy Protein) .................................. 3
1.3 大豆低聚糖(Soy Oligosaccharides) ........................ 4
1.4 大豆皂苷(Soy Saponin) .................................. 5
1.5 大豆異黃酮(Soy Isoflavones) ............................. 7
1.5.1 成分之結構與含量 ......................................... 7
1.5.2 物理與化學性質 .......................................... 10
1.6 大豆異黃酮之生理益處 ..................................... 10
1.7 大豆異黃酮萃取、純化分離方法 .............................. 13
1.7.1 萃取方法 ................................................ 13
1.7.2 純化分離方法............................................. 13
1.7.3 配糖型異黃酮之水解 ....................................... 14
1.7.3.1 酸水解 ................................................. 14
1.7.3.2 鹼水解 ................................................. 14
1.7.3.3 酶水解 ................................................. 15
1.7.3.4 酯化反應 ................................................ 15
1.8 大豆異黃酮分析方法 ....................................... 15
1.9 研究動機及目的 .......................................... 17
貳、最佳實驗製程 ................................................. 18
2.1 材料、藥品與儀器 ......................................... 18
2.1.1 原料 ................................................... 18
2.1.2 化學原料 ............................................... 18
2.1.3 儀器設備 ............................................... 18
2.2 分析方法 ............................................... 20
2.2.1 傅立葉紅外線光譜(FT-IR)定性分析 .......................... 20
2.2.2 紫外線-可見光光譜儀(UV-Vis)定性分析 ....................... 21
2.2.3 高效液相層析儀(HPLC)分析 ................................. 22
2.2.4 產率計算 ............................................... 24
2.3 最佳實驗製程 ............................................ 24
2.3.1 萃取步驟 ............................................... 26
2.3.2 純化與成分分離步驟 ....................................... 26
2.3.2.1 純化去糖型和配糖型異黃酮混合物 ............................ 26
2.3.2.2 純化Genistin ........................................... 28
2.3.2.3 純化配糖型異黃酮混合物 ................................... 31
2.3.3 大豆皂苷 ................................................ 33
2.3.4 製程所需材料設備與產率 ................................... 35
參、大豆萃取之製程選擇 ........................................... 37
3.1 製程方法選擇 ............................................ 37
3.1.1 熱水法(A) .............................................. 38
3.1.2 熱鹼水法(B) ............................................ 38
3.1.3 60 %甲醇法(C) .......................................... 38
3.2 結果討論 ................................................ 38
3.3 結論 ................................................... 39
肆、大豆蜜中分離去糖型和配糖型異黃酮混合物之製程選擇 ................. 40
4.1 製程方法選擇 ............................................ 40
4.1.1 溶劑法(A) .............................................. 41
4.1.2 酸沉澱法(B) ............................................ 41
4.1.3 樹脂法(C) .............................................. 42
4.2 結果討論 ................................................ 42
4.2.1 溶劑法(A) .............................................. 43
4.2.1.1 鹼式醋酸乙酯產物(A1) .................................... 43
4.2.1.2 酸式醋酸乙酯產物(A2) .................................... 43
4.2.2 酸沉澱法(B) ............................................ 44
4.2.3 樹脂法(C) .............................................. 45
4.2.3.1 HP 20 column(C1) ...................................... 46
4.2.3.2 HP 2MG column( C2) .................................... 46
4.3 結論 ................................................... 48
伍、Genistin分離及純化之製程選擇 .................................. 49
5.1 製程方法選擇 ............................................ 49
5.1.1 pH=12之NaOH脫附(A1) .................................... 49
5.1.2 pH=12之60%甲醇脫附(A2) .................................. 50
5.1.3 pH=3.0之HCl脫附(B1) .................................... 50
5.1.4 pH=3.0之60%甲醇脫附(B2) ................................. 50
5.2 結果討論 ................................................ 50
5.2.1 pH=12之NaOH脫附(A1) .................................... 50
5.2.2 pH=12之60%甲醇脫附(A2) .................................. 50
5.2.3 pH=3.0之HCl脫附(B1) .................................... 50
5.2.4 pH=3.0之60%甲醇脫附(B2) ................................. 51
5.3 結論 ................................................... 52
陸、總結 ........................................................ 53
參考文獻 ........................................................ 54
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