臺灣博碩士論文加值系統

近年來對活細胞氧化釋放的活性氧(Reactive oxygen species)的即時追蹤和定量越來越感興趣，因為它們是發病機理的潛在生物標記。本文描述了利用普魯士藍微立方體(PB MCs, Prussian blue Mircocube)修飾的石墨烯化奈米碳管(Graphenated carbon nanotubes, g-CNTs)的快速且靈敏的無酵素H2O2測定。我們通過簡單的水熱合成法製備PB MCs，並與g-CNTs混合，得到PB MCs / g-CNTs三維分層網狀結構的奈米複合材料。同時透過掃描式電子顯微鏡(Scanning electron microscope, SEM)、能量散佈X光分析儀(Energy-dispersive x-ray spectroscopy, EDX)，X光繞射分析儀(X-ray diffraction, XRD)，傅立葉轉換紅外光譜儀(Fourier transform infrared spectroscopy, FT-IR)驗證了複合材料的結構和組成。並以g-CNT / PB MCs薄膜修飾電極進行電化學檢測，其在最小的過電位值下顯示出優異的電催化活性使H2O2還原。由結果顯示，我們成功構建出一種具快速、靈敏、選擇性和可重複利用的電流傳感器，其有效的工作範圍為0.02-1935μM，最低檢測極限為5±1.2 nM，具有低過電位值、增強訊號、反應迅速和電極的穩定性等優點。最後在實際樣品檢測部分，除了成功地測定了人類血清中外加H2O2之外，也證明該方法可用於追蹤Raw 264.7哺乳動物細胞中的體內產生的H2O2。

Recent years have seen a growing interest towards real-time tracking and quantification of reactive oxygen species such as, H2O2 released in living cells, as they are potential biomarkers for pathogenesis. Herein, we described a rapid and sensitive enzymeless H2O2 assay using Prussian blue microcubes (PB MCs) decorated graphenated carbon nanotubes (g-CNTs) and this assay was successfully demonstrated for the tracking of in-vivo H2O2 production in Raw 264.7 mammalian cells. The PB MCs was prepared through facile hydrothermal route and blended with g-CNTs to yield 3D hierarchical network of PB MCs/g-CNTs nanocomposite. The structure and composition of the composite was verified by SEM, EDX, mapping, XRD, FT-IR, impedance and electrochemical methods. The g-CNTs/PB MCs film modified electrode was fabricated which displayed excellent electrocatalytic activity to H2O2 reduction at minimized overpotential. A rapid, sensitive, selective, durable and reproducible amperometric sensor was constructed that displayed working range of 0.02–1935 µM, and low limit of detection of 5±1.2 nM. The method was successful in the determination of H2O2 spiked in human serum. The other advantages of the method are low overpotential, enhanced signal, quick response time, practicality in mammalian cells and robustness of the electrode.

摘要 i
ABSTRACT ii
誌謝 iv
目錄 v
圖目錄 viii
表目錄 x
第一章 前言 1
第二章 文獻探討 2
2.1化學修飾電極 2
2.2生化感測器 2
2.3過氧化氫的生理關聯性 4
2.3.1簡介 4
2.3.2發炎反應 5
2.3.3 神經退化性疾病 7
2.4現今檢測過氧化氫的方法 8
2.4.1 螢光探針法 8
2.4.2 化學發光法 9
2.4.3 分光光度法 11
2.4.4 電化學分析法 12
2.5普魯士藍 13
2.5.1 簡介 13
2.5.2 生物感測器應用 13
2.6石墨烯化奈米碳管 16
2.6.1 石墨烯 18
2.6.2 奈米碳管 20
第三章 研究動機與分析方法 22
3.1研究動機 22
3.2實驗儀器 23
3.3特性分析方法 24
3.3.1掃描式電子顯微鏡觀察法 24
3.3.2 X光繞射分析法 25
3.3.3傅立葉轉換紅外線光譜法 26
3.3.4電交流阻抗分析法 27
3.4電化學偵測法 28
3.4.1 循環伏安法 28
3.4.2安培法 30
第四章 材料合成與實驗設計 31
4.1實驗藥品列表 31
4.2材料合成 32
4.2.1石墨烯化奈米碳管 32
4.2.2普魯士藍微立方體 32
4.3以g-CNTs/PB MCs薄膜修飾工作電極 33
4.4在RAW264.7真實樣本中即時監測與定量H2O2 33
第五章 結果與討論 34
5.1 g-CNTs, PB MCs與g-CNTs/PB MCs的SEM, EDX與Mapping分析 34
5.2 g-CNTs, PB MCs與g-CNTs/PB MCs的XRD和FTIR分析圖譜 35
5.3 g-CNTs/PB MCs的阻抗分析與電化學研究 36
5.4 g-CNTs/PB MCs/GCE對H2O2的電催化活性研究 38
5.5利用安培法對g-CNTs/PB MCs/GCE進行H2O2的測定 40
5.6 g-CNTs/PB MCs/GCE之選擇性、穩定性與再現性的研究 42
5.7 g-CNTs/PB MCs/GCE於RAW 264.7中對H2O2的即時監測 44
第六章 結論 46
參考文獻 47

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