臺灣博碩士論文加值系統

氧化碳超臨界萃取技術具有較低溫度，高擴散能力之特性，可將天然物中的熱不穩定性物質萃出，並保留活性成分，所以本研究以二氧化碳超臨界流體萃取技術萃取洛神花多酚類物質咖啡酸。洛神花所含有的多酚類物質具有許多的生物活性，其中的有效成分咖啡酸，具備有抗氧化、捕捉自由基、抑制酪氨酸酶的功能。高效能液相層析儀分析水萃取法、乙醇萃取法、二氧化碳超臨界流體萃取法對於洛神花的咖啡酸萃取效率。結果發現使用水萃取法與酒精萃取法均能將咖啡酸完全萃取出來，使用二氧化碳超臨界流體萃取法萃取出來的咖啡酸含量稍低，但是高效能液相層析儀的分析圖譜顯示使用二氧化碳超臨界流體萃取法的萃取物成分較為單純。另外將不同比例的水、乙醇作為共溶劑添加至二氧化碳超臨界流體中進行修飾，並且改變不同的臨界壓力、臨界溫度、與超臨界流體的流速藉此評估使用共溶劑修飾的超臨界流體萃取洛神花咖啡酸效率的最佳化條件。實驗結果發現，提高咖啡酸的萃取條件需要降低臨界壓力、提高臨界溫度、增加修飾共溶劑加入二氧化碳超臨界流體的流速、並且同時使用含有水與乙醇當作修飾共溶劑。根據高效能液相層析儀針對萃取物的咖啡酸含量分析結果，可以得到的最佳化萃取條件為臨界壓力250 bar、臨界溫度40℃、共溶劑乙醇濃度85%、共溶劑流量1.2 ml/min。總合全部的實驗結果，可以得知以水和酒精當作共溶劑進行修飾可以在二氧化碳超臨界流體中提高針對咖啡酸的萃取效率，但是水分含量過高容易使二氧化碳超臨界流體的狀態不穩定而形成次臨界流體，加入乙醇為修飾共溶劑則可以進一步穩定含有水分的二氧化碳超臨界流體，因此同時以兩種以上的共溶劑對二氧化碳超臨界流體進行修飾，是提高咖啡酸萃取效率的關鍵。在抗氧化活性當中，其氧化還原能力、清除DPPH自由基及酪氨酸酶之能力測定發現當咖啡酸萃取率上升，清除DPPH自由基則提升，而氧化還原能力上咖啡酸萃取率也頗佳，但酪氨酸酶能力上，則看不出相同趨勢，但在HPLC圖譜上較複雜，可能萃取出含有較優異抑制酪氨酸酶之成分。

Supercritical carbon dioxide (SC-CO2) can provide a benefit of lower temperature and high diffusion ability in nature product extraction process, not only preserved the thermo unstable but also keep the bioactivity of the extracts. Owing to this, we use supercritical carbon dioxide extract the caffeic acid (CA), a kind of polyphenol, from Roselle. Caffeic acid has abilities with antioxidant, radical catching and tyrosinase inhibition. We also use high performance liquid chromatography analysis the components differences between water extraction, alcohol extraction and supercritical carbon dioxide extraction. The result show that caffeic acid was complete extracted by water and alcohol extraction, but lower in supercritical carbon dioxide extraction. The components of supercritical carbon dioxide extraction are much simple. Following, we add water and alcohol as the co-solvent to modify the polarity of the supercritical carbon dioxide, and use this modified supercritical carbon dioxide in the caffeic acid extraction with differ critical temperatures, critical pressures and flow rates. In the results of the further analysis by high performance liquid chromatography, the best optimization in modified supercritical carbon dioxide extraction of caffeic acid needs critical pressure at 250 bar, critical temperature at 40℃, Co-solvent 85% ethanol as co-solvent at the flow rate 1.2ml/min. In conclusion, add water and alcohol as co-solvent into supercritical carbon dioxide can optimize the caffeic acid extraction efficiency from Roselle. Actually, add only water into supercritical carbon dioxide will make supercritical unstable and become sub-supercritical. Adding alcohol can provide a better stability when water is a necessary co-solvent to the supercritical carbon dioxide extraction. Both adding water and alcohol as co-solvents are the key point of increasing the efficiency of caffeic acid supercritical carbon dioxide extraction from Roselle.

中文摘要………………………………………………………………I
Abstract………………………………………………………………III
誌謝……………………………………………………………………V
目錄……………………………………………………………………VI
圖目錄………………………………………………………………IX
表目錄………………………………………………………………XII
符號說明……………………………………………………………XIII
第一章 緒論…………………………………………………………1
1.1 前言……………………………………………………1
1.2 研究動機與目的………………………………………5
第二章 文獻回顧…………………………………………………7
2.1 超臨界流體之簡介……………………………………7
2.1.1 超臨界流體之特性……………………………11
2.1.2 超臨界流體萃取技術之工業上應用…………16
2.1.3 二氧化碳超臨界流體萃取技術………………20
2.1.4 超臨界流體基礎理論…………………………26
2.1.5 二氧化碳超臨界流體萃取設備………………28
2.1.6 二氧化碳超臨界流體共溶劑選擇及添加……32
2.1.6-1 共溶劑修飾的種類…………………34
2.1.6-2 二氧化碳超臨界流體共溶劑添加方
式……………………………………35
2.2 洛神花之簡介…………………………………………37
2.3 多酚類化合物之簡介…………………………………42
2.4 咖啡酸(Caffeic acid) 之簡介…………………………44
第三章 實驗設備與方法………………………………………………45
3.1 實驗材料與藥品………………………………………45
3.2 實驗設備………………………………………………47
3.3 實驗架構及方法 ………………………………………53
3.3.1 超臨界二氧化碳萃取法與溶劑萃取法…………53
3.3.2 超臨界二氧化碳萃取法(不添加共溶劑) ………54
3.3.3 有機溶劑乙醇萃取法……………………………54
3.3.4 純水萃取法………………………………………55
3.3.5 高效能液相層析儀分析法(HPLC) ……………55
3.3.6超臨界萃取設備參數之改變以及氧化活性實驗58
3.3.7 二氧化碳超臨界流體萃取參數之改變…………59
3.4 抗氧化之活性測定……………………………………60
3.4.1 氧化還原能力(TPTZ)之測定……………………60
3.4.2 清除DPPH自由基能力之測定…………………60
3.4.3 酪氨酸酶(Tyrosinase)之測定……………………61
第四章 結果與討論……………………………………………………62
4.1 二氧化碳超臨界流體萃取法與傳統萃取法之比較……62
4.1.1 熱水萃取法……………………………………62
4.1.2 有機溶劑乙醇萃取法…………………………63
4.1.3 二氧化碳超臨界流體萃取法……………………63
4.2 二氧化碳超臨界流體萃取改變參數…………………69
4.2.1萃取臨界壓力改變之影響………………………71
4.2.2共溶劑流量改變之影響…………………………75
4.2.1共溶濃度改變之影響……………………………79
4.2.4萃取臨界溫度改變之影響………………………83
4.2.5 最佳萃取條件之選擇……………………………85
4.3 氧化活性之分析………………………………………87
4.3.1氧化還原能力(TPTZ)之能力……………………87
4.3.2清除DPPH自由基能力之能力…………………89
4.3.3酪氨酸酶(Tyrosinase)之能力……………………91
第五章 結論…………………………………………………………93
參考文獻……………………………………………………………… 95
附錄1 未添加共溶劑與最佳化萃取條件之咖啡酸效率比較………106
附錄2 標準品咖啡酸與各項Sample之HPLC圖譜………………106
附錄3 作者簡介及論文發表……………………………………125






圖目錄
Fig 1-1 咖啡酸Caffeic acid (CA)…………………………………6
Fig 2-1 物質之溫度與壓力關係示意相圖……………………………7
Fig 2-2不同狀態下分子間勢能與平均分子動能關係……………15
Fig 2-3 二氧化碳溫度對壓力之相圖………………………………21
Fig 2-4 超臨界流體之三相變化……………………………………22
Fig 2-5 超臨界流體萃取之基本操作圖……………………………29
Fig 2-6 洛神花………………………………………………………38
Fig 2-7 本研究所用之洛神花之花萼………………………………39
Fig 3-1二氧化碳超臨界流體萃取設備簡圖………………………49
Fig 3-2 二氧化碳超臨界萃取設備，金屬工業研究發展中心……50
Fig 3-3超臨界二氧化碳萃取與 ASES 法微粉成形實驗設備圖…51
Fig 3-4 第一部分實驗結構圖………….……………………………53
Fig 3-5 HPLC結構圖………………………………………………56
Fig 3-6 HPLC實體圖………………………………………………57
Fig 3-7第二部分實驗結構圖………………………………………58
Fig 4-1 咖啡酸 CA(Coffeic acid)標準品HPLC偵測圖譜………65
Fig 4-2 熱水萃取法HPLC偵測圖譜………………………………66
Fig 4-3 有機溶劑乙醇萃取法HPLC偵測圖譜……………………67
Fig 4-4二氧化碳超臨界萃取法(不添加共溶劑)HPLC偵測圖譜…68
Fig 4-5 共溶劑乙醇濃度95 %，臨界溫度為40 ℃，不同臨界壓力、
共溶劑流速之萃取條件，萃取物咖啡酸濃度分析曲線……72
Fig 4-6 共溶劑乙醇濃度90 %，臨界溫度為40 ℃，不同臨界壓力、
共溶劑流速之萃取條件，萃取物咖啡酸濃度分析曲線……73
Fig 4-7 共溶劑乙醇濃度85 %，臨界溫度為40 ℃，不同臨界壓力、
共溶劑流速之萃取條件，萃取物咖啡酸濃度分析曲線……73
Fig 4-8 共溶劑乙醇濃度95 %，臨界溫度為40 ℃，不同臨界壓力、
共溶劑流速之萃取條件，萃取物咖啡酸濃度分析曲線……76
Fig 4-9 共溶劑乙醇濃度90 %，臨界溫度為40 ℃，不同臨界壓力、
共溶劑流速之萃取條件，萃取物咖啡酸濃度分析曲線……77
Fig 4-10 共溶劑乙醇濃度85 %，臨界溫度為40 ℃，不同臨界壓力、
共溶劑流速之萃取條件，萃取物咖啡酸濃度分析曲線……77
Fig 4-11共溶劑乙醇流速0.6 ml/min，臨界溫度為40 ℃，不同臨界
壓力、共溶劑乙醇濃度之萃取條件，萃取物咖啡酸濃度分
析曲線………………………………………………………80
Fig 4-12 共溶劑乙醇流速0.9 ml/min，臨界溫度為40 ℃，不同臨界
壓力、共溶劑乙醇濃度之萃取條件，萃取物咖啡酸濃度分
析曲線………………………………………………………80
Fig 4-13 共溶劑乙醇流速1.2 ml/min，臨界溫度為40 ℃，不同臨界
壓力、共溶劑乙醇濃度之萃取條件，萃取物咖啡酸濃度分析
曲線…………………………………………………………81
Fig 4-14 共溶劑流速0.9 ml/min，共溶劑濃度95%，不同臨界溫度、
臨界壓力下之動態萃取曲線……………………………84
Fig 4-15 咖啡酸萃取效率與氧化還原力比較……………………88
Fig 4-16咖啡酸萃取效率與清除DPPH自由基能力比較…………90
Fig 4-17咖啡酸萃取效率與抑制酪氨酸酶能力比較………………92






















表目錄
Table 2-1 超臨界流體之重要發展史………………………..………10
Table 2-2 流體在不同狀態下之基本物理性質………………..……12
Table 2-3 常用的超臨界流體及臨界參數………………………..…13
Table 2-4 超臨界萃取技術應用於工業上之研究……………..……19
Table 2-5 常見之共溶劑與溶解度參數…………………………..…28
Table 2-6 分子極性的推測……………………………………..……33
Table 4-1各項萃取法之多酚類成分咖啡酸萃取結果………………65
Table 4-2 各項超臨界二氧化碳參數及產率活性結果………..……70
Table 4-3 超臨界萃取洛神花實驗數據之參數(改變壓力) ……..…74
Table 4-4 超臨界萃取洛神花實驗數據之參數(改變乙醇流量) ..…78
Table 4-5 超臨界萃取洛神花實驗數據之參數(改變乙醇濃度) …..82
Table 4-6 超臨界萃取洛神花實驗數據之參數(改變溫度) ……..…85
Table 4-7　洛神花粗取物超臨界操作參數與氧化還原力……..….…88
Table 4-8洛神花粗取物超臨界操作參數與清除DPPH自由基能力分
析……………………………………………………………………..…90
Table 4-9洛神花粗取物超臨界操作參數與抑制酪氨酸酶(Tyrosinase)
的能力分析…………………………………………………………..…92

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