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研究生: 柳承佑
Liu, Cheng-You
論文名稱: 以酪氨酸酶、殼聚醣、氧化還原石墨烯製備高靈敏度和選擇性的網印印刷碳電極用於多巴胺檢測
High Sensitivity and Selectivity Screen-Printed Carbon Electrode Fabricated by Tyrosinase, Chitosan, and Reduced Graphene Oxide for Detection of Dopamine
指導教授: 葉怡均
Yeh, Yi-Chun
學位類別: 碩士
Master
系所名稱: 化學系
Department of Chemistry
論文出版年: 2018
畢業學年度: 106
語文別: 中文
論文頁數: 60
中文關鍵詞: 網印印刷電化學生物感測器酪氨酸酶殼聚醣多巴胺氧化還原石墨烯
英文關鍵詞: screen-printed electrochemical biosensor, tyrosinase, chitosan, dopamine, reduced graphene oxide
DOI URL: http://doi.org/10.6345/THE.NTNU.DC.026.2018.B05
論文種類: 學術論文
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  • 多巴胺是人體中重要的神經傳遞物質,其對於帕金森氏症、阿茲海默症皆有重大的影響。在實驗中以酪胺酸酶 (tyrosinase)、殼聚醣 (chitosan)、氧化還原石墨烯 (reduced graphite oxide,rGO) 修飾於網印印刷碳電極 (Screen-printed carbon electrode,SPCE) 作為電化學生物感測器,用於多巴胺的檢測。並且針對抗壞血酸以及尿酸,這類在生物體內常見的干擾物,可以避免此感測器不被干擾而影響偵測誤判。利用循環伏安法測量證明了,所提出的電化學感測器的高靈敏度和選擇性,偵測極限為22 nM,並且與先前文獻相比,有較廣的線性範圍為0.4-8 μM和40-500 μM。此外,所提出的電極被應用於健康人體的尿液樣品時,取得了令人滿意的準確率,表示其適用於生理樣品中多巴胺的分析。

    Dopamine (DA) is an important neurotransmitter in human body. It has an great effect on Parkinson's disease and Alzheimer's disease. In this study, we fabricated the electrochemical biosensor of screen-printed carbon electrode (SPCE) modified with tyrosinase, chitosan and redox graphene oxide (rGO) and the sensor was used for the detection of DA without the interference from ascorbic acid or uric acid. We demonstrated the high sensitivity and selectivity of the electrochemical sensors with detection limit of 22 nM, and broad linear ranges of 0.4-8 μM and 40-500 μM by cyclic voltammetry. In addition, the proposed electrode exhibited a satisfactory recovery rate when it applied to healthy human urine samples, indicating that it is suitable for the analysis of DA in physiological samples.

    誌謝 I 中文摘要 II ABSTRACT III 目錄 IV 圖目錄 VII 表目錄 X 第一章 緒論 1 1 生物感測器 1 1-1-1 生物感測器介紹 1 1-1-2 電化學生物感測器 2 2 兒茶酚胺類神經傳遞物質 3 3 電化學 5 1-3-1 循環伏安法3 (cyclic voltammetry,CV) 6 1-3-2 三電極系統 8 1-3-3 工作電極 8 1-3-4 電化學沉積電極 9 1-3-5 網印印刷電極 (screen‐printed electrode,SPE) 9 4 網印印刷電極的修飾 11 1-4-1 金屬的修飾 11 1-4-2 電子傳遞媒介質的修飾 11 1-4-3 選擇性薄膜的修飾 13 1-4-4 電化學前處理法 14 1-4-5 氧化還原石墨烯 (reduced graphene oxide,rGO ) 修飾 15 1-4-6 殼聚醣修飾電極 17 5. 酵素的修飾 18 1-5-1 吸附 (adsorption) 修飾 19 1-5-2 共價鍵 (covalent bond) 修飾 19 1-5-3 化學交聯法 (chemical cross-linking) 21 1-5-4 基質包埋法 (entrapment) 22 6. 酪胺酸酶 (tyrosinase) 23 7. 多巴胺電化學生物感測器文獻回顧 24 8. 研究動機 26 第二章 實驗器材與方法 27 2-1實驗器材 27 2-2 實驗藥品 28 2-3 實驗設計 29 2-4 實驗方法 30 2-4-1 rGO 溶液配製 30 2-4-2 rGO 與 chitosan 溶液配製 30 2-4-3 玻璃基材的清洗及OTS修飾 30 2-4-4 chitosan/rGO 溶液與碳膠電極組裝 30 2-4-5 酵素的配製 31 2-4-6 固定酵素 31 2-4-7 在相同溶液下不同電極的電化學行為 31 2-4-8 pH 值的優化 32 2-4-9 生物電極的電化學活性對多巴胺的感測 32 2-4-10 不同掃描速率下的電化學反應 32 2-4-11 干擾物測試 32 2-4-12 穩定性測試 33 2-4-13 真實樣品的感測 33 第三章 實驗結果與討論 34 3-1 利用網印技術修飾玻璃基材表面 34 3-2 利用化學鍵結修飾tyrosinase/chitosan/rGO於SPCE 表面 34 3-2-1 掃描式電子顯微鏡(SEM)的觀測結果 35 3-2-2 衰減全反射傅利葉轉換紅外線光譜儀 (ATR-FTIR) 的鑑定 37 3-2-3 水接觸角觀察電極表面親疏水性 40 3-3 標準氧化還原系統下的循環伏安法圖 41 3-4 循環伏安法偵測多巴胺 42 3-4-1 四種電極系統對多巴胺電流感測值 42 3-4-2 生物複合材料電極對酸鹼值的影響 43 3-4-3 生物複合材料電極對多巴胺的感測 44 3-4-4 掃描速率與生物複合材料電極對多巴胺的變化 46 3-5 生物複合材料電極的選擇性與穩定性 47 3-5-1 選擇性 47 3-5-2 穩定性 49 3-6 生物複合材料電極對真實樣品的測量 50 3-7 與先前研究文獻之多巴胺電化學感測器比較 51 第四章 總結 52 參考資料 53

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