臺灣博碩士論文加值系統

本實驗的主要目的是利用超臨界二氧化碳萃取綠茶（龍井茶）中的機能性成分，並利用超臨界層析，希望能以一套完全潔淨的設備，分離純化出高附加價值的兒茶素。萃取方面，改變溫度、壓力、修飾劑種類以及流速等各種變因，探討出萃取效果最佳的操作條件，並將傳統萃取法與超臨界萃取法的產物做比較；層析方面，先使用HPLC作初步的分析，大致了解流動相將成分分離的機制後，再使用SFC分析，改變溫度壓力來調整二氧化碳的溶解度，並調整修飾劑種類及比例等各種變因，以得到最佳的分離效果。
在HPLC方面，使用的是逆相層析系統，流動相的最佳沖提比例為0.6％H3PO4：ACN：EA＝88：10.5：1.5，流速為1ml/min，使用的波長在280nm時會有最大的吸收值。
目前研究結果顯示，超臨界靜態萃取較動態萃取有更優良的萃取效率，主要抗氧化的兒茶素（EGCG）在3500psi之後才會慢慢被萃取出來，並隨著壓力的升高有增加的趨勢，溫度方面在60℃會有最優良的效果產生，且當修飾劑乙醇的流速提升至0.4ml/min，萃取率會有非常明顯的增加。
而SFC在210nm時有最佳吸收波長，且在50℃、2000psi、二氧化碳流速2ml/min以及修飾劑乙醇流速為0.5ml/min的狀態下可以成功的分離其中四種目標物質。
由於SFE與SFC皆使用修飾劑乙醇而得到最佳效果，因此可以初步的將兩個系統結合在一起，達到預期的目標。

In this thesis, a supercritical carbon dioxide dynamic extraction of functional compositions of green tea was applied, and the extracts were analyzed by supercritical fluid chromatography. The goal is to separate the highly value-added catechins from the green tea with a absolutely pure extraction facility.
In order to find the best operation condition, various factors such as temperature, pressure, different modifiers, and the speed of flows were discussed. Furthermore, the comparisons between the productions of traditional extraction and supercritical fluid extraction have been made; the HPLC and SFC analysis were applied respectively. The HPLC figures out the mechanism of the separation, meanwhile, different temperatures and pressures were set to adjust the solubility of carbon dioxide. Also, different modifiers were used to test the separation efficiency.
A reversed HPLC method was applied, and the best proportion for mobile phase is 0.6% H3PO4 : ACN : EA = 88 : 10.5 : 1.5 with 1 ml/min flow speed. The maximum absorbance is at 280 nm wavelength. According to parent literatures, the efficiency of static extraction is better than dynamic extraction. EGCG starts to be extracted at 3500 psi when the temperature is 60℃. Besides, the percentage of extraction is high when the flow speed of ethanol in the modifier is 0.4 ml/min. For SFC, the best absorbent wavelength is 210 nm, and four of the compositions could be separated at 50℃, 2000 psi with a 2 ml/min CO2 flow speed and 0.5 ml/min ethanol flow speed. Due to the result from the modifier, ethanol, is promising, the SFE and SFC could be combined to obtain the expected solution.

指導教授推薦書……………………………………………………………
口試委員會審定書…………………………………………………………
國科會授權書………………………………………………………………iii
誌謝…………………………………………………………………………iv
中文摘要……………………………………………………………………v
英文摘要……………………………………………………………………vi
目錄………………………………………………………………………vii
圖目錄………………………………………………………………………x
表目錄……………………………………………………………………xiii
第一章 緒論………………………………………………………………1
第二章 文獻回顧…………………………………………………………3
2.1茶葉簡介……………………………………………………………3
2.1.1茶葉的化學組成……………………………………………3
2.1.2兒茶素類對人體的生理效應………………………………5
2.1.3兒茶素類之化學結構式……………………………………6
2.1.4兒茶素的製造流程…………………………………………10
2.2超臨界流體………………………………………………………11
2.2.1超臨界流體簡介……………………………………………11
2.2.2超臨界流體萃取方式………………………………………17
2.2.3溶解度參數…………………………………………………17
2.2.4修飾劑效應…………………………………………………20
2.3層析法原理………………………………………………………21
2.3.1分配係數、容量因子與選擇因子…………………………24
2.3.2層析法之比較………………………………………………26
2.4超臨界層析（SFC）………………………………………………29
2.4.1超臨界層析儀的操作變因…………………………………31
第三章 實驗方法…………………………………………………………34
3.1實驗材料與藥品…………………………………………………34
3.2實驗儀器…………………………………………………………35
3.2.1超臨界流體萃取系統………………………………………35
3.2.2超臨界流體層析系統………………………………………36
3.2.3其他儀器……………………………………………………37
3.3實驗流程…………………………………………………………38
3.3.1實驗基本架構………………………………………………38
3.3.2茶葉萃取……………………………………………………39
3.3.3 HPLC層析…………………………………………………39
3.3.4校正曲線的繪製……………………………………………42
3.3.5超臨界二氧化碳層析………………………………………43
第四章 結果與討論………………………………………………………44
4.1 HPLC層析的結果………………………………………………44
4.1.1調整流動相…………………………………………………45
4.1.2咖啡因的分離………………………………………………48
4.2超臨界萃取………………………………………………………51
4.2.1傳統萃取法與SFE之比較…………………………………51
4.2.2超臨界靜態萃取與動態萃取………………………………54
4.2.3溫度與壓力的影響…………………………………………57
4.2.4修飾劑效應…………………………………………………61
4.2.5萃取時間的影響……………………………………………66
4.2.6茶葉種類的影響……………………………………………69
4.2.7與文獻比較的結果…………………………………………71
4.3 SFC分析的結果…………………………………………………74
4.3.1兒茶素標準品之分析結果…………………………………74
4.3.2不同管柱的比較……………………………………………87
4.3.3超臨界萃取物之分析結果…………………………………89
第五章 結論………………………………………………………………90
參考文獻…………………………………………………………………91

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