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研究生:林政雲
研究生(外文):Cheng-Yun Lin
論文名稱:以逆流層析分離幾丁寡醣可行性之探討
論文名稱(外文):A Feasibility Study of Chitooligosaccharides Seperation in Countercurrent Chromatography
指導教授:余艇
指導教授(外文):Tiing Yu
學位類別:碩士
校院名稱:國立交通大學
系所名稱:應用化學系
學門:自然科學學門
學類:化學學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:中文
論文頁數:66
中文關鍵詞:逆流層析雙水相幾丁寡醣
外文關鍵詞:countercurrent chromatographyaqueous-two phase solvent systemchitooligosaccharides
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目前用來分離幾丁聚醣的方法有高效能液相層析、毛細管電泳,但此兩種方法皆無法收集大量樣品。本研究以逆流層析儀(countercurrent chromatography)結合雙水相溶劑系統(aqueous two phase solvent system)或有機溶劑/水相系統分離並收集幾丁寡糖。試驗的條件包括,改變溶劑的組成、比例、濃度與pH值。其結果顯示,在雙水相溶劑系統中,此類分子量較小、極性較高的幾丁寡醣分子的分離效果不佳,即醣類分子大都留在上層聚乙二醇相( Polyethylene Glycol,PEG )中,儘管提高PEG的分子量、濃度,改變溶劑中所含鹽的種類(包括: 磷酸鹽、檸檬酸鹽)及其所佔的比例,或將溶劑系統的pH值降低,均無法將寡醣流析分離出,此可能是因為幾丁寡醣分子與聚乙二醇間的親和力較大所造成的。而在有機溶劑/水相系統中,無論如何提高溶劑系統的極性或pH值,亦無法提高寡糖分子在兩相間的分配係數,即醣類分子均留在水相當中。
Two methods which are used to separate chitooligosaccharides are HPLC and CE, but they cannot handle great sample volume either. In this study, we try to separate and collect chitooligosaccharides in either of aqueous two phase solvent system and organic/aqueous solvent system in countercurrent chromatography. Experimental factors are composition of solvent, sample concentration and pH value. As the result, in aqueous two phase solvent system, chitooligosaccharides with low molecular weight and high polarity are hardly separated while they almost stay in the upper layer, PEG(polyethylene glycol) - rich phase. Even though we increase the molecular weight or concentration of PEG, add different kinds of salts including phosphate and citrate, change the concentration of salt, or lower the pH value in solvent system, chitooligosaccharides cannot be eluted out. It is suggested that a good affinity exists between chitooligosaccharides and PEG. In organic/aqueous solvent system, however, no matter how greatly we try to increase the polarity or the pH value of our solvent systems, the partition coefficient of chitooligosaccharides between the two phase cannot be increased, so they remain in the aqueous phase.
第一章 緒論……………………………………………………… 1
第二章 研究背景與理論………………………………………… 3
2.1逆流層析……………………………………………………. 3
2.1.1 簡介………………………………………………… 3
2.1.2 起源與發展………………………………………… 3
2.1.3 基礎理論…………………………………………… 4
2.1.4 儀器系統…………………………………………… 6
2.1.5 影響分離效果的因素……………………………… 13
2.1.6 環形螺旋離心式逆流層析………………………… 14
2.1.7 逆流層析的應用…………………………………… 17
2.2 雙水相溶劑系統……………………………………… 18
2.2.1 簡介………………………………………………… 18
2.2.2 理論………………………………………………… 19
2.2.3 相系統的性質……………………………………… 21
2.2.4 影響分配係數的變因……………………………… 22
2.3 偵測法…………………………………………………… 26
2.3.1 醣類的分析…………………………………………… 26
2.3.2 直接偵測……………………………………………… 27
2.3.3 間接偵測……………………………………………… 29
2.4 幾丁寡醣…………………………………………………… 31
2.4.1 來源與化學構造……………………………………… 31
2.4.2 幾丁寡醣的基本性質………………………………… 33
2.4.3 幾丁寡醣的應用……………………………………… 34
第三章 實驗……………………………………………………… 37
3.1 藥品與溶劑………………………………………………… 37
3.1.1 藥品…………………………………………………… 37
3.1.2 標準品………………………………………………… 38
3.1.3 溶劑…………………………………………………… 38
3.2 儀器裝置…………………………………………………… 38
3.3. 實驗步驟…………………………………………………… 40
第四章 結果與討論………………………………………… 42
4.1 雙水相溶劑系統…………………………………………… 42
4.1.1 實驗參數設定………………………………………… 43
4.1.2 間接偵測共競離子的選擇…………………………… 44
4.1.3 改變聚乙二醇的分子量……………………………… 45
4.1.4 改變聚乙二醇的濃度………………………………… 46
4.1.5 改變鹽的種類與比例………………………………… 48
4.1.6 改變pH值……………………………………………… 50
4.2 有機/水 溶劑系統………………………………………… 56
4.3 溶劑與儀器上的問題……………………………………… 59
第五章 結論……………………………………………………… 61
參考文獻…………………………………………………………… 62
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