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研究生：	鄭易修 Cheng, I-Hsiu
論文名稱：	以共價合成單體以製作模版高分子對膽紅素之電化學感測分析 Synthesis of functional monomer via covalent bondeing for the fabrication of imprinted polymer electrode for the electrochemical sensing of bilirubin
指導教授：	許梅娟 Syu, Mei-Jywan
學位類別：	碩士 Master
系所名稱：	工學院 - 化學工程學系 Department of Chemical Engineering
論文出版年：	2010
畢業學年度：	98
語文別：	中文
論文頁數：	73
中文關鍵詞：	膽紅素、分子模版高分子、電化學感測
外文關鍵詞：	bilirubin, imprinted polymer, electrochemical
相關次數：	點閱：79 下載：0
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膽紅素為血紅蛋白降解之產物，正常人每日可以生成 250 ~ 350 mg 的膽紅素，而醫學檢測中，總膽紅素的濃度的標準值為 0.2 ~ 1.5 mg/dL，而直接膽紅素的標準值小於 0.4 mg/dL，利用兩者之間的差值為間接性膽紅素的濃度，並藉以膽紅素濃度的判斷是否肝機能出現異常，而人體內總膽紅素濃度若大於 2 mg/dL 則會出現黃疸症狀。醫學上的檢測重氮法檢測總膽紅素含量，而以氧化法檢測直接膽紅素含量，方法較為費時且不方便。因應生物感測器微小化的發展，利用對膽紅素分子具特異性結合之高分子薄膜，並同時利用電化學感測的方式來偵測膽紅素濃度。
製備對膽紅素分子模版高分子的材料選用上，乃是以 MAA、histidine (His) 自行合成單體 MAA-His，搭配交聯劑 EGDMA (ethylene glycol dimethylacrylate) 進行自由基加成聚合反應，並與模版分子膽紅素產互補作用力，於高分子結構中留下具有模印效果的辨識性孔洞。本研究亦探討不同單體與模版分子之間的作用、預聚合液體積的影響以及不同電極基材之應用，並利用 SEM、potentiostat 分析 MIP、NIP 薄膜之表面構形及感測訊號之影響。
依據不同的電化學感測條件所對應之結果分析，poly((MAA-His)-co-EGDMA) 電極的靈敏度為 2.96 (μA/cm2)/(mg/dL)，模印因子為 5.879 ± 0.305，偵測線性範圍在膽紅素濃度 3 mg/dL 以下，對膽紅素與對應膽綠素的選擇性為 1.75 ± 0.076。研究數據顯示模板高分子可與電化學電流式結合偵測膽紅素。

Imprinting of bilirubin, a significant biomarker for hyperbilirubia, was accomplished by the co-polymerization of a synthesized monomer, MAA-His with EGDMA crosslinker. MAA-His was first proposed in this work and was prepared by coupling to achieve the formation of covalent bond between methacrylic acid (MAA) and histidine (His). Normally, to achieve a precise control on the formation of specific binding cavity site for the absorption of bilirubin, a large proportion of EGDMA was applied. Heated polymerization was used in the following experiments because of its rapid and easy implementation. The pre-polymerization solution containing bilirubin as the template was dropped and spread uniformly onto the Au surface. After the accomplishment of polymerization, proper porogen was applied to extract the imbedded bilirubin templates from the polymer matrix so that the binding cavity was formed in the matrix. The imprinting effect from different amounts of pre-polymerization solution was under investigation. Imprinting effect from the addition of MWCNTs into the pre-polymerization solution was also discussed. Sensitivity as well as selectivity of the fabricated imprinted polymer/Au toward bilirubin was obtained. The electrochemical performance of the imprinted sensing film for the specific sensing of bilirubin was also evaluated from several aspects such as limit of detection (LOD), repeatability, stability, and clinical feasibility from serum samples.

中文摘要........................................................................................................................ I
Abstract .........................................................................................................................II
誌謝..............................................................................................................................III
目錄............................................................................................................................. IV
表目錄........................................................................................................................ VII
圖目錄.......................................................................................................................VIII
第一章 緒論..................................................................................................................1
1-1 分子模版高分子 (Molecularly imprinted polymer, MIP) ............................1
1-1-1 原理介紹.............................................................................................1
1-1-2 製備流程.............................................................................................2
1-1-3 分子模版高分子的組成.....................................................................3
1-1-3-1 模版..........................................................................................3
1-1-3-2 功能性單體..............................................................................3
1-1-3-3 交聯劑......................................................................................4
1-1-3-4 溶劑..........................................................................................4
1-1-4 模印型態.............................................................................................5
1-1-4-1 非共價性模印..........................................................................5
1-1-4-2 共價性模印..............................................................................5
1-1-4-3 共價與非共價式模印..............................................................5
1-1-5 聚合方式.............................................................................................6
1-1-6 分子模版高分子近期發展.................................................................6
1-2 生物化學感測器............................................................................................8
1-2-1 生化感測元件 (Biological sensing element) .....................................8
1-2-2 生化感測分析.....................................................................................8
1-2-2-1 光學式感測器..........................................................................8
1-2-2-2 質量式感測器..........................................................................8
1-2-2-3 電化學感測器 (Electrochemical sensor)................................9
1-3 分子模版高分子與電化學的結合與應用..................................................10
1-3-1 表面模印高分子層...........................................................................10
1-3-2 電聚合高分子層...............................................................................10
1-3-3 表面修飾...........................................................................................10
1-4 膽紅素 (Bilirubin) .......................................................................................11
1-4-1 膽紅素的生成...................................................................................11
1-4-2 膽紅素在血液中的輸送...................................................................12
1-4-3 黃疸症狀 (Jaundice) ........................................................................13
1-4-4 膽紅素結構.......................................................................................14
1-4-5 膽紅素檢測方法...............................................................................16
1-5 研究動機......................................................................................................19
第二章 實驗方法與材料............................................................................................20
2-1 膽紅素的測定..............................................................................................20
2-1-1 呈色法...............................................................................................20
2-2-1 在氧化鋁板上濺鍍金層...................................................................22
2-2-2 奈米碳管的酸化...............................................................................22
2-2-3 膽紅素 MIP 製備............................................................................22
2-2-3-1 Poly(AMPS-co-EGDMA) 模印高分子.................................22
2-2-3-2 Poly(vinylimidazole-co-EGDMA) 模印高分子....................23
2-2-3-3 Poly(MAA-histidine-co-EGDMA) 模印高分子...................23
2-3 電化學分析..................................................................................................27
2-3-1 定電位 I-t 曲線圖...........................................................................27
2-3-2 選擇性測試與血清測試...................................................................27
2-4 實驗藥品目錄..............................................................................................28
2-5 實驗儀器目錄..............................................................................................29
第三章 結果與討論....................................................................................................30
3-1 分子模版高分子材料探討..........................................................................30
3-1-1 FT-IR 圖譜分析................................................................................31
3-1-2 電化學感測分析...............................................................................35
3-2-2 電極改質之電流分析.......................................................................39
3-3 不同單體的分子模版高分子......................................................................40
3-3-1 以 AMPS 螯合 imidazole 製備poly(AMPS-EGDMA) 模版高分子
......................................................................................................................40
3-3-2 以 vinylimidazole 為單體製備 poly(vinylimidazole-EGDMA) 模版高
分子..............................................................................................................42
3-3-3 以 MAA-histidine 作為單體製備 poly((MAA-His)-EGDMA) 模版高
分子..............................................................................................................44
3-3-4 不同單體的總比較...........................................................................46
3-4 分子模版高分子對膽紅素感測之探討......................................................49
3-4-1 改變模版的濃度...............................................................................49
3-4-2 添加濃度的變化測試.......................................................................50
3-4-3 電化學感測的可逆性測試...............................................................54
3-5 選擇性與干擾性測試..................................................................................57
3-5-1 膽紅素與膽綠素的選擇性...............................................................57
3-5-2 血清干擾性感測...............................................................................64
第四章 結論................................................................................................................68
參考文獻......................................................................................................................70
                                    

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