臺灣博碩士論文加值系統

本研究以三種共溶劑添加形態的超臨界二氧化碳，萃取甲魚卵粉中膽固醇與游離胺基酸，並以應答曲面實驗設計，探討各萃取的變因，影響膽固醇移除率、游離胺基酸萃出率、油脂萃出量、膽固醇濃縮因子、游離胺基酸濃縮因子的最適值。首先，經由索式乙醇萃取16小時，每克甲魚卵粉得到4.61毫克的膽固醇，索氏水16小時萃取，每克甲魚卵粉可得到10.55毫克的游離胺基酸。接著，以328K與250bar為設計中心點的乙醇修飾超臨界二氧化碳進行萃取。實驗結果顯示，膽固醇移除率，隨著萃取溫度及壓力上升。在338K與300bar時，靜態萃取有58wt%，動態萃取有70%的最大膽固醇移除率出現。
以308K與300bar為設計中心點的水修飾連續式二氧化碳萃取，經敏感度測試後，實驗結果顯示，當壓力為350bar與溫度308K時，游離胺基酸萃出率可達58%，純度由1.05%提昇至33.59%。在乙醇修飾連續萃取的結果顯示，膽固醇移除率，呈現持續上升的趨勢。當WP/WCS值為1/1.3及壓力350bar與溫度338K時，膽固醇移除率可達90%，萃出油中膽固醇濃度由0.46%上升至3.7%。

This study examined co-sovent modified supercritical carbon dioxide extraction of cholesterol and free amino acids from soft-shell turtle fish egg powder. Response Surface Methodology (RSM) was then applied to determine operation conditions in obtaining the optimal responses of cholesterol removing, the free amino acids recovery as well as concentration factors of cholesterol and free amino acids in the extracted oil. The content of cholesterol in the egg powder is 4.61 (mg/g), obtained by 16hour Soxhlet ethanol extraction. The content of free amino acids is 10.55 (mg/g), obtained by 16hour water Soxhlet extraction.
Center composite RSM schemes designed at 328K and 250bar for ethanol modified supercritical carbon dioxide extractions, presented the cholesterol removing rate was still increasing as temperature and pressure increased. The cholesterol removing rate was only up to 58 % for static SC-CO2 extraction and 70% for dynamic SC-CO2 extraction at 328K, 300bar, within the temperature and pressure investigated.
Water modified conutious supercritical carbon dioxide extraction further indicated that temperature and pressure are two major factors to enhance the recovery and purity of free amino acids in the extracted oil, while the recovery of free amino acids increased to 58%, and the purity of AA increased from 1.05% to 33.59%. at 350bar and 308K. Ethanol modified conutious supercritical carbon dioxide extraction of cholesterol at 350bar and 338K indicated that pressure is dominant. The removing efficiency of cholesterol attained 90%, relative concentration of cholesterol increased from 0.46% to 3.7% in the extracted oil.

摘要 I
Abstract II
目錄 IV
表目錄 VI
圖目錄 VIII
符號表 XI
第一章 緒論 1
1-1 甲魚簡介 1
1-2 甲魚成分及功效 2
第二章 文獻回顧 5
2-1 胺基酸種類與分析 5
2-2 膽固醇萃取與分析 8
2-3 卵黃油製備法 11
2-4 甲魚之功能性 12
2-5 超臨界二氧化碳原理與應用 15
研究動機 17
研究目的及方法 18
第三章 實驗方法與步驟 19
3-1 原料 19
3-2 藥品 19
3-3 儀器與設備 22
3-4 甲魚卵粉之樣品萃取 25
3-5 應答曲面(RSM)的實驗設計法 28
3-6 樣品分析與計算 30
第四章 結果與討論 35
4-1 索氏溶劑萃取製程 35
4-2靜態超臨界流體萃取乙醇修飾的卵粉 36
4-2-1 潤濕時間與乙醇添加比例對膽固醇移除率的影響 36
4-2-2 萃取時間對膽固醇移除率的影響 37
4-2-3 溫度與壓力對膽固醇移除率的影響 37
4-2-4 溫度與壓力對膽固醇初始移除速率的探討 38
4-3動態超臨界流體萃取乙醇修飾的卵粉 39
4-3-1 二氧化碳流速對膽固醇移除率的影響 39
4-3-2 二氧化碳用量對膽固醇移除率的影響 40
4-3-3 溫度與壓力對膽固醇移除率的影響 40
4-4連續式超臨界流體萃取共溶劑修飾的卵粉 41
4-4-1 乙醇添加對萃取膽固醇與游離胺基酸的影響 41
4-4-2 水添加對萃取膽固醇與游離胺基酸的影響 41
4-5 實驗設計法探討超臨界二氧化碳萃取膽固醇與胺基酸 42
4-5-1靜態超臨界流體萃取乙醇修飾的卵粉 42
4-5-2動態超臨界流體萃取乙醇修飾的卵粉 44
4-5-3連續式超臨界流體萃取共溶劑修飾的卵粉 45
第五章 結論 49
參考文獻 51

表1、索氏溶劑萃取20克卵粉中膽固醇與游離胺基酸的實驗數據 60
表2、靜態二氧化碳萃取乙醇修飾卵粉之膽固醇實驗數據 61
表3、靜態二氧化碳萃取乙醇修飾卵粉之膽固醇初始移除速率數據 62
表4、動態超臨界二氧化碳萃取乙醇修飾卵粉之膽固醇實驗數據 63
表5、連續式超臨界二氧化碳萃取共溶劑修飾卵粉之膽固醇與游離胺基酸的實驗數據 64
表6、靜態二氧化碳萃取乙醇修飾卵粉之應答曲面實驗設計數據 65
表7、動態二氧化碳萃取乙醇修飾卵粉之應答曲面實驗設計數據 66
表8、連續式二氧化碳萃取乙醇修飾卵粉之應答曲面實驗設計數據 67
表9、連續式二氧化碳萃取水修飾卵粉之應答曲面實驗設計數據 68
附表1、超臨界流體萃取天然物有效成分的代表文獻 91
附表2-1、HPLC分析游離胺基酸之溶媒梯度條件 92
附表2-2、膽固醇與游離胺基酸標準品配置在甲醇溶液之HPLC分析檢量線數據 93
附表2-3、膽固醇與游離胺基酸標準品配置在基質溶液(Soxhlet)的HPLC分析檢量線數據 94
附表2-4、膽固醇與游離胺基酸標準品配置在基質溶液(SCF)的HPLC分析檢量線數據 95
附表3、添加共溶劑之二氧化碳操作條件與臨界狀態 96
附表4、靜態超臨界二氧化碳萃取乙醇修飾之7克卵粉，膽固醇移除率之變方分析與二次回歸方程式 97
附表5、靜態超臨界二氧化碳萃取乙醇修飾之7克卵粉，總萃出量之變方分析與二次回歸方程式 98
附表6、動態超臨界二氧化碳萃取乙醇修飾之7克卵粉，膽固醇移除率之變方分析與二次回歸方程式 99
附表7、動態超臨界二氧化碳萃取乙醇修飾之7克卵粉，總萃出量之變方分析與二次回歸方程式 100
附表8、連續式超臨界二氧化碳萃取乙醇修飾之7克卵粉，膽固醇移除率之變方分析與二次回歸方程式 101
附表9、連續式超臨界二氧化碳萃取乙醇修飾之7克卵粉，游離胺基酸回收率之變方分析與二次回歸方程式 102
附表10、連續式超臨界二氧化碳萃取乙醇修飾之7克卵粉，總萃出量之變方分析與二次回歸方程式 103
附表11、連續式超臨界二氧化碳萃取乙醇修飾之7克卵粉，βCh值之變方分析與二次回歸方程式 104
附表12、連續式超臨界二氧化碳萃取水修飾之7克卵粉，膽固醇移除率之變方分析與二次回歸方程式 105
附表13、連續式超臨界二氧化碳萃取水修飾之7克卵粉，游離胺基酸回收率之變方分析與二次回歸方程式 106
附表14、連續式超臨界二氧化碳萃取水修飾之7克卵粉，βAA值之變方分析與二次回歸方程式 107

圖1、索式溶劑迴流萃取甲魚卵粉的裝置簡圖 69
圖2、超臨界二氧化碳萃取甲魚卵粉的裝置簡圖 70
圖3、溶劑萃取實驗，甲魚卵粉中膽固醇及游離胺基酸的萃出量與時間的關係 71
圖4、預潤濕時間與潤濕量影響靜態二氧化碳的膽固移除率 72
圖5、萃取時間對靜態超臨界二氧化碳移除膽固醇的關係 73
圖6、恆壓時，靜態超臨界二氧化碳萃取乙醇修飾之卵粉，溫度對膽固醇移除率的影響 74
圖7、恆溫時，靜態超臨界二氧化碳萃取乙醇修飾之卵粉，壓力對膽固醇移除率的影響 75
圖8、靜態超臨界二氧化碳萃取乙醇修飾之卵粉，壓力與溫度對膽固醇初始移除率的影響 76
圖9、動態超臨界二氧化碳萃取乙醇修飾之卵粉，CO2使用量對膽固醇移除率的影響 77
圖10、恆壓時，動態超臨界二氧化碳萃取乙醇修飾之卵粉，溫度對膽固醇移除率的影響 78
圖11、恆溫時，動態超臨界二氧化碳萃取乙醇修飾之卵粉，壓力對膽固醇移除率的影響 79
圖12、靜態超臨界二氧化碳萃取乙醇修飾之卵粉，膽固醇移除率與變數因子之等高線與立體應答曲面圖 80
圖13、靜態超臨界二氧化碳萃取乙醇修飾之卵粉，總萃出量與變數因子之等高線與立體應答曲面圖 81
圖14、動態超臨界二氧化碳萃取乙醇修飾之卵粉，膽固醇移除率與變數因子之等高線與立體應答曲面圖 82
圖15、動態超臨界二氧化碳萃取乙醇修飾之卵粉，總萃出量與變數因子之等高線與立體應答曲面圖 83
圖16、連續超臨界二氧化碳萃取乙醇修飾之卵粉，膽固醇移除率與變數因子之等高線與立體應答曲面圖 84
圖17、連續超臨界二氧化碳萃取乙醇修飾之卵粉，游離胺基酸回收率與變數因子之等高線與立體應答曲面圖 85
圖18、連續超臨界二氧化碳萃取乙醇修飾之卵粉，總萃出量與變數因子之等高線與立體應答曲面圖 86
圖19、連續超臨界二氧化碳萃取乙醇修飾之卵粉，βCh值與變數因子之等高線與立體應答曲面圖 87
圖20、連續超臨界二氧化碳萃取水修飾之卵粉，膽固醇移除率與變數因子之等高線與立體應答曲面圖 88
圖21、連續超臨界二氧化碳萃取水修飾之卵粉，游離胺基酸回收率與變數因子之等高線與立體應答曲面圖 89
圖22、連續超臨界二氧化碳萃取水修飾之卵粉，βAA值與變數因子之等高線與立體應答曲面圖 90
附圖1、20種胺基酸的分子結構圖 108
附圖2、膽固醇的結構圖 111
附圖3、特定流體之壓力與溫度相圖 112
附圖4、膽固醇之HPLC分析圖 113
附圖5、17種胺基酸之HPLC分析圖 114
附圖6、甲魚卵粉經萃取之外觀變化 115
附圖7、ASP之三種基質簡量線 116
附圖8、GLU之三種基質簡量線 116
附圖9、SER之三種基質簡量線 117
附圖10、THR之三種基質簡量線 117
附圖11、GLY之三種基質簡量線 118
附圖12、ALA之三種基質簡量線 118
附圖13、ARG之三種基質簡量線 119
附圖14、PRO之三種基質簡量線 119
附圖15、VAL之三種基質簡量線 120
附圖16、MET之三種基質簡量線 120
附圖17、ILE之三種基質簡量線 121
附圖18、LEU之三種基質簡量線 121
附圖19、PHE之三種基質簡量線 122
附圖20、CYS之三種基質簡量線 122
附圖21、LYS之三種基質簡量線 123
附圖22、HIS之三種基質簡量線 123
附圖23、TYR之三種基質簡量線 124
附圖24、膽固醇之三種基質簡量線 124

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