臺灣博碩士論文加值系統

中 文 摘 要
本研究係利用功能性四氧化三鐵-聚乙烯亞胺奈米微粒 (Fe3O4–PEI) 結合電子傳遞介質–赤血鹽與葡萄糖氧化酵素 (glucose oxidase, GOD)，以滴覆乾燥的方式修飾於網印電極上 (screen-printed elec- trodes, SPEs)，進而製備成電流式葡萄糖酵素感測電極。文中利用循環伏安法 (cyclic voltammetry, CV) 及計時安培法 (chronoamperometry, CA) 針對 SPEs 之修飾過程及電化學特性進行分析。結果顯示：經由Fe3O4–PEI修飾的電極，其氧化還原反應呈現準可逆與典型的擴散控制行為，且Fe3O4–PEI會提升電流訊號，是因為它的高親水性與可以在生物分子中扮演奈米級的電極。此葡萄糖生物感測器亦呈現快速應答、高靈敏度 (約 0.33�n�嫀/mM)、以及寬廣之線性應答濃度 (可達1,000 mg/dl)。

Abstract
An amperometric glucose biosensor was successfully constructed by simply drop-coating iron oxide-polyethylenimine (Fe3O4-PEI) and ferri- cyanide (as a mediator) onto screen-printed electrodes (SPEs), and then layering-on glucose oxidase (GOD). The electrochemical characteristics of the modified SPEs were analyzed by cyclic voltammetry (CV), electro- chemical impedance spectroscopy (EIS) and chronoamperometry (CA). Results showed that the redox reaction at Fe3O4-PEI modified SPEs was quasi-reversible and demonstrated a typical diffusion-limited behavior. In addition, Fe3O4-PEI could enhance the response current of glucose biosensor because of its high degree of hydrophilicity and acting like nanoscaled electrodes for the biomolecules. The glucose biosensors also exhibit a relatively fast response and high sensitivity (ca. 0.33�n�嫀/mM) with a wide linear concentration range up to 1000 mg/dl of glucose.

目 錄
頁次
中文摘要 I
英文摘要 II
目 錄 IV
表 目 錄 VII
圖 目 錄 VIII
符號說明 IX

第 一 章 緒論 1
1.1 前言 1

第 二 章 文獻回顧與研究動機 4
2.1 葡萄糖生物感測器的發展史 4
2.2 電子傳遞介質 5
2.3 網印電極 (screen-printed electrodes, SPEs) 6
2.4 奈米顆粒修飾性生物感測器 8
2.5 磁性奈米顆粒修飾性生物感測器 9
2.6 polyethylenimine (PEI) 於生物感測器之應用 11
2.7 實驗動機與目的 17

第 三 章 電化學理論與分析方法 19
3.1 電化學理論 19
3.1.1 電化學反應 19
3.1.2 法拉第定律 19
3.1.3 電雙層與電極-溶液界面 21
3.2 電化學分析方法 23
3.2.1 循環伏安法 (cyclic voltammetry, CV) 23
3.2.2 計時安培法 (chronoampermetry, CA) 25
3.2.3 電化學阻抗分析法 (electrochemical impedance spectro- scopy,
EIS) 26

第 四 章 材料與實驗方法 32
4.1 藥品與材料 32
4.2 儀器與設備 32
4.2.1 量測儀器 32
4.2.2 其他設備 33
4.3 實驗架構
34
4.4 實驗步驟 35
4.4.1 Fe3O4-PEI 奈米微粒之製備 36
4.4.2 評估 Fe3O4-PEI 奈米微粒在網印電極上之電化學特性 37
4.4.3 Ferri 和Ferri/Fe3O4-PEI 奈米微粒之掃描速率的動力學參數 38
4.4.4 Ferri/Fe3O4-PEI 奈米微粒之葡萄糖感測器的最適化條件之
Ferri/Fe3O4-PEI 奈米微粒之覆蓋量 39
4.4.5 Ferri/Fe3O4-PEI奈米微粒葡萄糖感測器的最適化條件之電位 40
4.4.6 Ferri/Fe3O4-PEI奈米微粒葡萄糖感測器的最適化條件之酵素滴覆量 41
4.4.7 Ferri/Fe3O4-PEI奈米微粒葡萄糖感測器的最適化條件之pH 42
4.4.8 比較Ferri/GOD、Ferri-Fe3O4/GOD、Ferri/PEI-Fe3O4/GOD修飾性
SPEs 於葡萄糖感測器之檢量線 43

第 五 章 結果與討論 45
5.1 評估Fe3O4-PEI磁性奈米微粒在網印電極上之電化學特性 45
5.1.1 評估Fe3O4-PEI磁性奈米微粒在網印電極上無電子傳遞介質下之電化學
特性 45
5.1.2 評估Ferri/Fe3O4-PEI磁性奈米微粒在網印電極上之電極修飾過程的界
面現象 47
5.1.3 評估Ferri/Fe3O4-PEI奈米微粒在葡萄糖氧化酵素網印電極上的催化特
性 50
5.2 Ferri 和 Ferri/Fe3O4-PEI 奈米微粒之改變不同掃描速率並探討動力
學參數 52
5.3 Ferri/Fe3O4-PEI 奈米微粒於葡萄糖感測器的最適化條件 55
5.3.1 最適化Ferri/Fe3O4-PEI覆蓋量 55
5.3.2 最適化電位 57
5.3.3 最適化酵素滴覆量 59
5.3.4 最適化pH 61
5.4 比較Ferri/GOD、Ferri/Fe3O4/GOD、Ferri/Fe3O4-PEI/GOD 修飾性
SPEs 於葡萄糖感測器之檢量線 63
5.6 Ferri/Fe3O4-PEI/GOD 修飾性SPEs與其他奈米微粒修飾性電極之葡萄
糖生物感測器之性質比較 66

第 六 章 結論 68

參考文獻 69


































表目錄
頁次
表 2.1 利用不同修飾物修飾四氧化三鐵奈米微粒並加以運用於不同領域 12
表 2.2 利用不同修飾物修飾四氧化三鐵奈米微粒在生物感測器上的應用 13
表 5.1 修飾性SPEs的動力學參數 53
表 5.2 不同修飾性 SPEs 葡萄糖感測器性質之比較 65
表 5.3 不同奈米微粒修飾性葡萄糖生物感測器之性質探討 67
















圖目錄
頁次
圖 1.1 生物感測器的示意圖 2
圖 2.1 第一代葡萄糖生物感測器反應機制圖 4
圖 2.2 第二代葡萄糖生物感測器的反應機制圖 6
圖 2.3 網印電極示意圖 7
圖 2.4 polyethylenimine (PEI) 結構圖 15
圖 3.1 電雙層示意圖 22
圖 3.2 循環伏安法原理示意圖；(a) 電位控制，(b) 電流響應 23
圖 3.3 計時安培法原理示意圖；(a) 電位階躍，(b) 電流響應 27
圖 3.4 (a) 電極與溶液的介面及整體溶液 (b) 電子電路圖 28
圖 3.5 典型的等效電路圖形 29
圖 3.6 經修正後的等效電路圖 30
圖 3.7 典型的奈式圖形 30
圖 4.1 研究架構流程圖 34
圖 5.1 循環伏安圖：(a) Fe3O4-PEI修飾性 SPEs滴入PBS (pH = 7，0.1M )測試；(b)
Fe3O4-PEI/GOD 修飾性 SPEs滴入葡萄糖溶液 (200mg/dl) 測試。操作條件：掃
描速率為 50 mV/s。 46
圖 5.2 循環伏安圖：(a) Ferri 修飾性 SPEs；(b) Ferri/GOD 修飾性 SPEs；(c)
Ferri/Fe3O4-PEI 修飾性 SPEs；(d) Ferri/Fe3O4-PEI/ GOD 修飾性 SPEs。分
別滴入PBS (pH = 7，0.1 M) 測試。操作條件：掃描範圍為－0.8∼＋0.8 V；
掃描速率為 50 mV s-1。 49
圖 5.3 循環伏安圖：(a)(b) Ferri/Fe3O4-PEI/GOD 修飾性 SPEs分別滴入PBS (pH =
7、0.1 M) 與滴入葡萄糖溶液 (200 mg/dl) 測試。操作條件：掃描速率為 50
mV/s。 51
圖 5.4 循環伏安圖：(A) Ferri修飾性SPEs；(B) Ferri/Fe3O4-PEI 修飾性 SPEs
分別滴入PBS (pH = 7，0.1 M) 測試，掃描速率由 (a) 至 (h) 分別為: 5 、
10、20、40、60、100、150、200 mV/s。(A)(B)嵌入之圖為掃描速率開根號對波
峰電流的作圖。 54
圖 5.5 Fe3O4-PEI奈米微粒添加量對計時安培法 (CA) 之影響 56
圖 5.6 不同的應用電位對計時安培法 (CA) 之影響 58
圖 5.7 不同的酵素滴覆量對計時安培法 (CA) 之影響 60
圖 5.8 不同的 pH 之葡萄糖溶液對計時安培法 (CA) 之影響 62
圖 5.9 不同的Ferri-GOD、Ferri-Fe3O4/GOD、Ferri/Fe3O4-PEI/GOD 修飾性SPEs以不同
葡萄糖濃度依計時安培法 (CA) 進行檢量線實驗 65

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