離子液體應用於脂溶性蛋白之快速萃取及檢測

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姓名

王執仲(Zhi-Zhong Wang) 查詢紙本館藏

畢業系所

生醫科學與工程學系

論文名稱

離子液體應用於脂溶性蛋白之快速萃取及檢測
(Ionic liquid used for rapid extraction and detection of liposoluble protein)

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摘要(中)

本研究論文進行一系列離子液體之合成與分析並將其應用於生物脂溶性蛋白之萃取與檢測測試。脂溶性蛋白之萃取、純化與濃縮，在生物醫學相關研究與實際應用上是相當重要的工程問題，傳統生化工程上萃取分離脂溶性蛋白的方式有反微胞萃取與雙水相溶劑工程等方式，但其程序與效率仍有需多可改善之空間。離子液體具有低溶點、高熱穩定性、高極性、耐強酸與低蒸氣壓等特殊性質，並可根據所需的物理或化學特性改變其陰陽離子之組合與變化以進行難溶性分子之溶解，此外，離子液體具有安全可回收等特性，可有效降低環境之汙染，有機會取代現行常用之有機溶劑。因此，本研究採用了離子液體並針對脂溶性蛋白之萃取與純化進行相關應用與分析。
在合成與驗證部分，本研究論文首先針對一系列不同結構之離子液體進行合成與分析，並探討不同濃度和結構下的離子液體對於脂溶性蛋白之溶解能力。研究結果中顯示[1-MIM-Pentane][OMs-] 對麩質蛋白有最佳的溶解能力，在室溫下，1毫升的40％ [1-MIM-Pentane][OMs-] 的離子液體可溶解737ppm的麩質蛋白。此外，在驗證部分也利用合成之離子液體以麩質蛋白為例結合免疫磁珠與磁電化學系統進行脂溶性蛋白的快速萃取檢測應用，在整體檢測上僅需數分鐘內既可成功完成脂溶性蛋白萃取與檢測分析，所測得之線性度達99%，LOD為2 ppm相較於一般的酵素免疫分析法，大幅減少樣品前處理時間並有效提升檢測速度。
本研究論文成功完成一系列離子液體之合成與分析並以麩質蛋白為例成功將其應用於生物脂溶性蛋白之萃取與檢測測試。本研究成果除了提供未來在脂溶性蛋白液相萃取相關技術上之方式之選擇外，也拓展了離子液體之相關應用層面。

摘要(英)

In this research carries out the synthesis and analysis of a series of ionic liquids and applies them to the extraction and detection of biological liposoluble protein. The extraction, purification, and concentration of liposoluble protein are very important engineering issues in biomedical research and applications. The methods of extracting and separating liposoluble protein in traditional biochemical engineering include reverse microcell extraction and aqueous two-phase solvent engineering. However, those methods are still for improvement in its procedures and efficiency. Ionic liquids have special properties such as low melting point, high thermal stability, high polarity, resistance to strong acids and low vapor pressure. Ionic liquids can change the combination and change of anions and cations according to the required physical or chemical characteristics to dissolve insoluble molecules. Also, ionic liquids are safe and recyclable, which can effectively reduce environmental pollution and have the opportunity to replace the current commonly used organic solvents. Therefore, in this study, ionic liquids were used for liposoluble protein extraction and purification. Then the results were used to application and analysis.
In the synthesis and verification section, this research first synthesized and analyzed a series of ionic liquids with different structures, and explored the solubility of ionic liquids with different concentrations and structures to liposoluble protein. The results show that [1-MIM-PEG550][OMs-] has the best solubility for fat-soluble gluten protein. At room temperature, 1 ml of 40% [1-MIM-PEG550][OMs-] ionic liquid can dissolve 855ppm fat-soluble gluten protein. Also, in the verification part, the synthetic ionic liquid is used to take the fat-soluble gluten protein as an example, combined with the immune micromagnetic beads and the electrochemical system for rapid extraction and detection of liposoluble protein. In the overall detection, it only takes a few minutes to finish the extraction and detection of liposoluble protein. The correlation coefficient is 0.99 and the limit of detection is 12.6 pg/mL. Compared with the general enzyme-linked immunosorbent assay (ELISA) it greatly reduces the time of sample preparation and effectively improves the detection speed.
This research completed the synthesis and analysis of a series of ionic liquids and took the fat-soluble gluten protein as an example to successfully apply it to the extraction and detection of biological liposoluble protein. This research not only provide the technique of liposoluble protein liquid phase extraction in the future choices, but also expanded the application level of ionic liquids.

關鍵字(中)

★ 離子液體
★ 脂溶性蛋白
★ 微米磁珠
★ 電化學
★ 麩質蛋白

關鍵字(英)

★ Ionic liquid
★ Liposoluble protein
★ Magnetic beads
★ Electrochemistry
★ Gluten

論文目次

中文摘要 i
誌謝 iv
目錄 v
圖目錄 vii
表目錄 viii
專有名詞縮寫對照表 ix
符號說明 x
第一章緒論 1
1-1 前言 1
1-2 文獻回顧 2
1-3 研究動機和目的 4
1-4 論文架構 4
1-5 藥品與儀器 5
1-5-1 藥品 5
1-5-2 儀器 6
第二章離子液體 7
2-1 背景 7
2-2 離子液體合成 8
2-2-1 [X][OMs-]離子液體合成流程 9
2-2-2 [X][Cl-]離子液體合成流程 10
2-3 以核磁共振光譜儀驗證離子液體之合成 11
2-3-1 NMR樣品製備流程 12
2-3-2 離子液體之1-H NMR 12
2-4 離子液體溶解麩質蛋白定量分析 16
2-4-1 麩質蛋白樣品製備與量測 21
2-4-2 量測結果與分析 21
第三章微米磁珠 23
3-1 背景 23
3-2 微米磁珠合成 24
3-2-1 合成策略 24
3-2-2 奈米磁性粒子合成流程 29
3-2-2 矽殼層生成 30
3-2-3 MCB-silatrane修飾微米磁珠 31
3-3 微米磁珠粒徑分析 31
3-3-1 穿透式電子顯微鏡 31
3-3-2 穿透式電子顯微鏡樣品製備 32
3-3-2 粒徑分析結果 32
3-4 磁珠XPS分析 35
3-4-1 背景與原理 35
3-4-2 XPS樣品製備 35
3-4-3 磁珠XPS結果與分析 36
3-5 固定生物分子於微米磁珠 37
3-5-1 背景與原理 37
3-5-2 樣品製備 38
第四章脂溶性蛋白於電化學系統檢測 40
4-1 電化學背景與原理 40
4-2 電化學系統操作流程與參數設定 45
4-3 電化學系統檢測麩質蛋白濃度之檢量線 46
第五章結論 47
參考文獻 49

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指導教授

黃貞翰(Chen-Han Huang)

審核日期

2020-10-5

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