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論文名稱: | 市售魚油的品質調查及以固定化石蓴脂氧合酶修飾魚油氣味的研究
| A Survey on Quality of Commercial Fish Oil and Improvement of Its Aroma Using Immobilized Algal Lipoxygenases
研究生: | 劉誌成 Chih-Cheng Liu
指導教授: | 孫寶年 Bonnie Sun Pan
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: | m92320019
畢業學年度: | 93
語文別: | 中文
關鍵字: | 魚油 fish oil
| 脂氧合酶 lipoxygenase
| 氣味 aroma
全文點閱次數: | 25

全文影像連結:

論文全文(pdf)

論文頁數: | 92
摘要: | 魚油中的魚腥味主要來自多元不飽和脂肪酸 ( PUFA ) 之氧化，為了增進魚油的應用性，本研究針對市售食用魚油進行脂肪酸組成與氣味品質分析，再以固定化海藻脂氧合酶修飾魚油氣味，期增加魚油之應用性。
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| 魚油以脂肪酸甲酯化法中反應條件較激烈的BF3 ( boron trifluoride ) 及反應條件較緩和的TMAH ( tetramethylammonium hydroxide ) 酯化後，分別進行氣相層析。由於BF3法以熱迴流進行轉酯化，可將魚油中TG及乙酯化態的EPA與DHA轉為甲酯化態，而TMAH法在室溫反應，僅能甲酯化TG型態的EPA與DHA，經層析圖比較後，顯示美國產魚油樣品膠囊中含TG及乙酯化態之EPA與DHA兩種型態。
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| 以BF3酯化法分析，市售食用魚油膠囊之EPA與DHA總合佔其脂肪酸總量之15-75 %。市售魚油膠囊內之魚油具魚腥味，氣味強度差異甚大。另添加保健成分之複方魚油，其添加物有銀杏、大蒜萃取物、紅麴萃取物、植物醇、卵磷脂、γ-胺基丁酸等，其魚油的腥氣較單方魚油強。
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| 石蓴脂氧合酶修飾魚油氣味，自2004年8月至2005年4月葉狀體製成粗酵素液，其脂氧合酶比活性範圍在1.14 - 2.90 μmol/mg-min。颱風後採集之石蓴，LOX活性高達4.95 μmol/mg-min。採集退潮後水面下之石蓴，其活性範圍介於1 - 2.90 μmol/mg-min。石蓴脂氧合酶以硫酸銨部分純化，在飽和濃度40 - 60 % 的劃分範圍具有活性，在透析液中添加0.2 mM Ca2+可以提升酵素的活性，回收率為59.72 %。
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| 石蓴粗酵素液及經過硫酸銨飽和濃度40-60 % 部分純化之石蓴LOX，分別與載體oxirane acrylic beads混合，於室溫中75 rpm震盪20小時進行固定化處理。固定化之石蓴LOX與0.5μmole亞麻油酸作用於25℃、以75 rpm 震盪反應，反應需2小時達平衡，酵素活性高於固定化之石蓴粗酵素液。固定化之石蓴LOX與市售膠囊魚油於25℃、75 rpm震盪反應2小時後，發現有好的氣味產生，這些氣味分子的化學結構以GC/MS鑑定，為甜瓜味 ( Ethyl acetate )、水果味 ( E,E-3,5-Octadiene-2-one ) 等。
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論文目次: | 表目錄    x
| 圖目錄    xii
| 貳、文獻回顧    3
| 一、魚油的特性    3
| 1.脂肪酸組成    3
| 2.安定性    3
| 3.營養價值    4
| 二、油脂氣味的形成    5
| 1.脂肪酸的氧化    5
| 2.氧化產物    6
| 3.揮發性成分    7
| 三、氣味分析方法    8
| 四、脂氧合酶與水產品氣味的關係    14
| 1.酵素與魚油作用後的氣味    14
| 2.修飾氣味的應用    14
| 五、脂氧合酶固定化方式與特性    15
| 1.固定化方式    15
| 2.固定化LOX的特性    17
| 參、材料與方法    19
| 一、材料    19
| 1.市售魚油    19
| 2.海藻    19
| 3.Oxirane acrylic beads    19
| 二、分析方法    19
| 1.石蓴脂氧合酶酵素液之製備    19
| (1)緩衝溶液    19
| (2)石蓴脂氧合酶粗酵素液萃取    20
| (3)硫酸銨分劃    20
| 2.脂氧合酶活性測定 - 吸光度法 ( LOOH 產物測定 )    21
| 3.蛋白質定量 - 染料結合法 ( dye binding )    22
| 4.固定化酵素的製備 - 化學共價鍵法    22
| 5.固定化脂氧合酶活性之測定    23
| 6.固定化酵素反應時間測定    23
| 7.魚油修飾後之香氣品評    23
| 8. 揮發性化合物鑑定- Headspase Solid Phase Microextraction法    24
| (1)氣味化合物之萃取    24
| (3)氣液相層析-質譜儀GC/MS分析    25
| (4)滯留指數 ( retention index, R.I. ) 測定    25
| 9.脂肪酸分析    27
| (1)甲基酯化    27
| (2)氣相層析分析    28
| 肆、結果與討論    29
| 一、市售魚油脂肪酸之組成    29
| 1. 魚油產品的分類    29
| 2. 魚油脂肪酸之組成    39
| 二、石蓴脂氧合酶之固定化    44
| 1. 石蓴脂氧合酶之活性    44
| 2. 脂氧合酶之部分純化    47
| 3. 固定化石蓴脂氧合酶    51
| 三、石蓴脂氧合酶對魚油氣味之修飾    53
| 1. 石蓴香氣化合物之鑑定    53
| 2. 魚油氣味化合物之鑑定    56
| 3. 修飾魚油之脂肪酸組成與氣味鑑定    62
| 陸、結論    68
| 柒、參考文獻    70
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