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研究生： 張巍耀
論文名稱： 利用漩渦輔助液液微萃取結合螢光高效率液相層析儀偵測高濃度胺基酸樣品中的微量脂肪胺
Vortex-assisted liquid-liquid microextraction combined with HPLC for the fluorometric determination of trace aliphatic amines in water samples with high levels of amino acids
指導教授： 吳劍侯
口試委員: 林萬寅
黃賢達
鄧金培
吳劍侯
學位類別： 碩士
Master
系所名稱： 原子科學院 - 生醫工程與環境科學系
Department of Biomedical Engineering and Environmental Sciences
論文出版年： 2011
畢業學年度： 99
語文別： 中文
論文頁數： 42
中文關鍵詞： 漩渦輔助液液微萃取法 、鄰苯二甲醛 、高效率液相層析儀 、螢光 、脂肪胺
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    • 利用漩渦輔助液液微萃取法(Vortex-assisted liquid-liquid microextraction)結合螢光高效率液相層析儀分析高濃度胺基酸樣品中的微量直鏈脂肪胺並避免胺基酸的干擾。本論文探討了乳化方式、萃取溶劑選擇、萃取溶劑體積、螢光放射波長、vortex乳化時間、離心時間及萃取時間等變數，其方法線性範圍接近兩個數量級且有良好的線性(R2 > 0.99)，C1至C8直鏈胺的偵測極限介於0.09-0.31 nM，相對標準偏差介於1.9-9.1%，擁有不錯的偵測極限及精密度。最後將此方法應用到市售的優酪乳、啤酒及運動飲料中C1到C8直鏈胺的量測，並監測銅胺基酸光照樣品中所產生的胺產物。

    目錄 中文摘要.....I 英文摘要.....II 目錄.....III 圖目錄.....VI 表目錄.....VIII 附錄目錄.....i 第一章 前言.....1 1.1 簡介.....1 1.2 研究目的.....1 1.3 原理及方法選擇.....2 第二章 文獻回顧.....3 2.1 o-Phthaldialdehyde/2-mercaptoethanol螢光衍生法.....3 2.2 傳統萃取法.....4 2.2.1 固相萃取法(Solid-phase extraction).....4 2.2.2 液相萃取法(Liquid-phase extraction).....4 2.3 微萃取法.....5 2.3.1 固相微萃取法(Solid-phase microextraction).....5 2.3.2 液相微萃取法(Liquid-phase microextraction).....6 2.3.2.1 單滴液相微萃取法(Single drop microextraction).....6 2.3.2.2 液液液三相微萃取法(Liquid-liquid-liquid microextraction).....7 2.3.2.3 應用中空纖維之液相微萃取.....9 2.3.2.4 分散式液液微萃取法(Dispersive liquid-liquid microextraction; DLLME).....10 第三章 實驗方法.....13 3.1 實驗裝置.....13 3.1.1 單光光照系統.....13 3.1.2 高效率液相層析儀(High performance liquid chromatography; 簡稱HPLC).....14 3.1.3 離心機.....14 3.1.4 漩渦混合機.....14 3.2 實驗藥品.....14 3.3 藥品、試劑製備.....15 3.3.1 硼酸緩衝液製備.....15 3.3.2 磷酸緩衝液製備.....16 3.3.3 衍生試劑製備.....16 3.3.4 C1-C8直鍊胺母液製備.....16 3.3.5 21種胺基酸(包括β-alanine)混合母液製備.....17 3.3.6 NaCl母液製備.....17 3.3.7 Cu(II)母液製備.....17 3.3.8 β-Alanine母液製備....17 3.3.9 醋酸鈉沖提液製備.....17 3.4 實驗步驟.....17 3.4.1 標準溶液.....17 3.4.2 真實樣品.....18 3.4.2.1 優酪乳樣品.....18 3.4.2.2 啤酒樣品.....19 3.4.2.3 銅胺基酸光照樣品.....19 3.4.2.4 運動飲料樣品.....20 第四章 結果與討論.....20 4.1 萃取溶劑的選擇.....20 4.2 萃取溶劑體積.....25 4.3 螢光放射波長.....26 4.4 Vortex乳化時間.....27 4.5 離心時間.....28 4.6 分析方法的表現.....30 4.7 真實樣品分析.....31 4.7.1 優酪乳和啤酒樣品.....31 4.7.2 銅胺基酸光照樣品.....34 4.7.3 運動飲料樣品.....35 第五章 結論.....36 第六章 未來展望.....36 參考文獻.....37 附錄.....i 圖目錄 圖1.3.1 聚丙烯多孔中空纖維截面圖.....3 圖2.1 OPA/2-ME衍生試劑與胺反應機制圖.....4 圖2.3.1 固相微萃取(SPME)裝置.....5 圖2.3.2.1.1 單滴液相微萃取裝置.....6 圖2.3.2.1.2 斜口針懸掛溶劑液滴近照.....7 圖2.3.2.2.1 液液液三相微萃取法裝置.....8 圖2.3.2.2.2 液液液三相微萃取法透試圖.....8 圖2.3.2.3.1 中空纖維液相微萃取法裝置.....9 圖2.3.2.3.2 溶液棒液相微萃取法裝置.....10 圖2.3.2.4.1 分散式液液微萃取法流程圖.....11 圖2.3.2.4.2 勻相液液萃取法流程圖.....12 圖3.1.1 單光光照系統.....13 圖4.1.1 使用chloroform萃取溶劑之HPLC圖譜.....21 圖4.1.2 使用醇類萃取溶劑之HPLC圖譜.....22 圖4.1.3 使用isooctane萃取溶劑之萃取前後的HPLC圖譜.....23 圖4.1.4 使用烷類萃取溶劑之HPLC圖譜.....24 圖4.1.5 烷類萃取溶劑的選擇.....25 圖4.2 萃取溶劑體積的影響.....26 圖4.3 螢光放射波長的影響.....27 圖4.4 Vortex時間的影響.....28 圖4.5 離心時間的影響.....29 圖4.6 萃取時間的影響.....30 圖4.7.1.1 優酪乳樣品經isooctane萃取前後及添加20 nM胺類標準品的HPLC圖譜.....32 圖4.7.1.2 啤酒樣品1經isooctane萃取前後的HPLC圖譜.....33 圖4.7.2.1 不同光照時間下的Cu(II)-β-alanine錯合物樣品稀釋10倍後再經isooctane萃取的HPLC圖譜.....34 圖4.7.2.2 Cu(II)-β-alanine錯合物光化學反應機制.....35 圖4.7.3 運動飲料經isooctane萃取前後的HPLC圖譜.....35 表目錄 表3.4.1 HPLC沖提之時間梯度.....18 表4.5 分析參數一覽.....29 表4.6.1 分析方法的表現.....30 表4.6.2 其它偵測脂肪胺的方法比較.....31 表4.7.1市售優酪乳及啤酒樣品量測結果.....33 附錄目錄 A. 乳化方式的比較.....i B. 萃取時間影響.....i C. C1-C8直鏈胺檢量線.....ii D. 濃縮因子和不同衍生條件下兩相強度比較.....iii

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