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研究生: 周芳妃
Fang-Fei Chou
論文名稱: 電化學壓電晶體液體感測器研製與應用
Preparation and Application of Chemical Electrode Piezoelectric Crystal Liquid Sensors
指導教授: 施正雄
Shih, Jeng-Shong
學位類別: 博士
Doctor
系所名稱: 化學系
Department of Chemistry
論文出版年: 2008
畢業學年度: 96
語文別: 中文
論文頁數: 184
中文關鍵詞: 電化學電極表面聲波轉能器石英晶體微天平鄰近電場金屬離子碳六十固定化酵素葡萄糖氧化酶膽固醇水解酶
英文關鍵詞: electrochemical electrode, surface acoustic wave, transducer, quartz crystal microbalance, fringing electric field, metal ions, fullerene, immobilized enzyme, glucose oxidase, cholesterol esterase
論文種類: 學術論文
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  • 本研究發展一套電極/表面聲波元件(ESAW)系統將超高頻波UHF應用於水溶液化學分析。ESAW系統使用ESAW表面聲波元件為ST-cut石英表面聲波感測元件,此系統應用於水中各種離子濃度偵測及葡萄糖水溶液定量工作。ESAW系統優點是製作成本非常低廉,組裝方法非常容易,而且長時間偵測不會發生訊號能量衰弱現象。建構ESAW系統方法是基於干擾ST-cut石英SAW震盪元件石英晶體表面鄰近電場以產生訊號,製作時以長距離導線及同軸線將放置於水溶液中的各種電極連接到石英SAW震盪元件的金屬外殼。ESAW系統偵測的陽離子包含鹼金族、鹼土族及過渡金屬元素。未經前濃縮步驟,此系統直接測得Cu2+離子偵測極限為7.6 ppm,其靈敏度為2.55 × 105 Hz/(mol/L)。此系統偵測空白訊號標準偏差為10 Hz,信度為99.86 %。ESAW系統亦發展做為葡萄糖生化感測器,其以葡萄糖氧化酶(GOD)催化葡萄糖氧化反應。另外,ESAW系統也應用於葡萄糖與過氧化氫混合溶液系統。ESAW葡萄糖生化感測器在低於10-3M葡萄糖溶液中可測得斜率為9.3 × 102 Hz decade−1 (Hz/logM)的檢量線,且其靈敏度高於QCM葡萄糖生化感測器。
    另外,本研究也以AT-cut石英塗佈碳六十C60大環胺醚方式發展QCM微天平之長鏈脂肪酸與膽固醇酯生化感測器。在非離子型界面活性劑的乳化液中以QCM感測器偵測長鏈脂肪酸或膽固醇酯水解產物。進行膽固醇酯水解反應之酵素添加方式包括兩種方式:在待測溶液中加入膽固醇水解酶以及放置碳六十C60固定化膽固醇水解酶濾片。此感測器系統測得膽固醇偵測極限為62 μM,此與目前醫學上光譜分析法偵測極限比較結果是良好的。目前醫學上光譜分析法仍需使用三種酵素反應系統,但本研究研發的膽固醇感測器只需應用使用一種酵素反應系統,再加上碳六十固定化酵素技術,因此大幅降低目前膽固醇定量分析的成本。

    An electrochemical electrode/surface acoustic wave (ESAW) system was developed to explore the possibility of the application of UHF waves, 300–3000MHz for chemical analysis in solution. The ESAW system with a ST-cut surface-acoustic wave (SAW)/quartz transducer was prepared for detection of metal ions and glucose in aqueous solutions. The ESAW system has the advantages of very low cost, easy fabrication and detection without quick energy-loss. The ESAW system was an on-line detection system and it was built-up by the interference on the fringing electric field of the ST-cut SAW quartz resonator. A set of electrodes welded with long-distance wires and coaxial cables was used to contact to the metal shell of the 315MHz SAW quartz resonator. The ESAW system was applied to detect various metal ions, e.g., alkaline metal, alkaline-earth metal and transition-metal ions. Without pre-concentration technique, the detection limit of Cu2+ ion with the ESAW detection system was estimated to be 1.2 × 10−4 mol L-1 (i.e. 7.6 ppm, from an analytical sensitivity of 2.55 × 105 Hz/(mol/L) and the standard deviation of the blank signal of 10 Hz with a confidence level of 99.86 %. The ESAW detection system was also applied as a biosensor for glucose to detect the glucose oxidation reaction by glucose oxidase (GOD) in aqueous solutions. The glucose oxidase (GOD) enzyme-catalyzed system was also studied on the detection of glucose / H2O2 mixture. The glucose ESAW biosensor with glucose oxidase exhibited a linear frequency response to the log concentration of glucose with a slope of approximately 9.3 × 102 Hz decade−1 (Hz/logM). The ESAW detection system also showed a good selectivity and a good detection limit of < 10−3M for glucose in aqueous solution. Furthermore, the ESAW detector showed much more sensitive than QCM crystal sensor for glucose.
    QCM crystal sensors for long-chain fatty acid and the cholesterol ester were also built up by using the AT-cut quartz crystal with fullerene C60-cyptand-22 coating. The QCM crystal sensors detected the long-chain fatty acid and the cholesterol ester concentration in a non-ionic surfactant emulsion solution. The hydrolysis of cholesterol ester was carried out with catalysts of free and fullerene C60-immobilized cholesterol esterase, respectively. The detection limit of cholesterol with the QCM crystal sensors was estimated to be 62 μM in good comparison with the clinical spectroscopic method. The clinical spectroscopic method is a tri-enzyme reactions system with very expensive cost but the QCM cholesterol sensors with fullerene-immobilized cholesterol esterase was a mono-enzyme system with low cost substantially for the quantitative measurement of cholesterol.

    英文摘要 1 中文摘要 3 目錄 5 圗目錄 8 表目錄 11 第一章 緒論 12 1-1 碳六十自組裝薄膜化學 12 1-2 碳六十壓電感測器 19 1-3 表面聲波元件 32 1-4 通訊電磁波輻射與電磁干擾 41 第二章 ESAW系統的建立 49 2-1 前言 49 2-2 實驗部份 53 2-2-1 儀器設計 53 2-2-2 藥品及試劑 59 2-2-3 實驗步驟 59 2-2-3.1 ESAW系統的硬體設計的研發 59 2-2-3.2 偵測電極的製作 59 2-2-3.3 偵測水溶液性質的基本實驗流程 63 2-3 結果與討論 64 2-3-1 ESAW系統的硬體設計 64 2-3-2 建立ESAW系統的硬體參數 71 2-3-2.1 導線長度效應 71 2-3-2.2 電源電壓效應 74 2-3-2.3 電極面積效應 76 2-3-2.4 電極間的距離效應 78 2-3-2.5 電極種類影響 81 2-3-2.6 溶液溫度效應 85 2-3-2.7 溶液介電性質效應 87 第三章 ESAW系統在離子水溶液的感測應用 91 3-1 前言 91 3-2 實驗部份 92 3-2-1 儀器 92 3-2-2 藥品及試劑 92 3-2-3 實驗步驟 93 3-3 結果與討論 94 3-3-1 強電解質水溶液的偵測 94 3-3-1.1 陽離子之偵測 94 3-3-1.2 陰離子之偵測 99 3-3-1.3 離子之偵測極限 101 3-3-1.4 ESAW系統在電解質水溶液中的延伸應用 102 3-3-2 ESAW系統在電解質水溶液中的理論探討 106 3-3-2.1 離子移動率的效應 106 3-3-2.2 不同濃度硝酸鉀水溶液訊號之預測 109 3-3-2.3 ESAW系統在強電解質水溶液中訊號干擾機制 110 第四章 ESAW系統/過氧化氫/葡萄糖感測器 113 4-1 前言 113 4-2 實驗部份 114 4-2-1 儀器 114 4-2-2 藥品及試劑 114 4-2-3 實驗步驟 115 4-3 結果與討論 116 4-3-1 C/Ag-型電極催化H2O2分解反應 116 4-3-2 酵素催化H2O2分解反應 120 4-3-3 ESAW延遲時間和H2O2/葡萄糖濃度之關係 123 4-3-4 葡萄糖水溶液的偵測 126 第五章 碳六十固定化酵素脂肪酸及膽固醇酯壓電感測器 133 5-1 前言 133 5-2 實驗部份 135 5-2-1 儀器 135 5-2-2 藥品及試劑 135 5-2-3 實驗步驟 136 5-2-3.1 C60-Cryptand-22與QCM電極之製備 136 5-2-3.2 QCM偵測系統之設計 137 5-2-3.3 膽固醇酯類的乳化液之配製 141 5-2-3.4 碳六十固定化酵素製作與保存 142 5-3 結果與討論 145 5-3-1 油酸濃度的測定 145 5-3-1.1 非離子型界面活性劑的濃度效應 145 5-3-1.2 QCM電極的C60-Cryptand-22塗佈效應 149 5-3-2 膽固醇酯濃度的測定 154 5-3-3 碳六十固定化膽固醇水解酶的測試實驗 164 第六章 結論 170 參考資料 174 圖1-1 碳六十之反應性 4 圖1-2 碳六十在基材表面形成化學吸附薄膜之方式 5 圖1-3 碳六十在金(Au)表面形成薄膜之自組裝反應 5 圖1-4 在金(Au)表面的碳六十排列方式 6 圖1-5 碳六十在矽氧化物表面形成薄膜之自組裝反應 6 圖1-6 碳六十固定化脂肪酵素濾片的裝置 7 圖1-7 生化感測器的構成與原理 10 圖1-8 生化感測器的生化辨識元與轉換器感測元 11 圖1-9 生化感測器的構成與原理 13 圖1-10 石英壓電晶體生化感測器的硬體設計 18 圖1-11 碳六十—血紅素塗佈於鉭酸鋰表面聲波元件 19 圖1-12 表面聲波壓電晶體生化感測器的硬體設計 19 圖1-13 生化感測器的發展潛能 20 圖1-14 聲波能量與縱深距離之關係 22 圖1-15 幾種常見的表面聲波型態 23 圖1-16 指叉型電極在石英晶片上產生表面聲波 24 圖1-17 表面聲波IDTs電極設計示意圖 26 圖1-18 Rayleigh-SAW與液體接觸示意圖 27 圖1-19 各式感測器中的表面聲波元件 29 圖1-20 電磁波頻譜 31 圖1-21 無線通訊電波範圍 33 圖1-22 導線四周空間的電場輻射方式 36 圖1-23 無所不在的電磁波的干擾現象 36 圖2-1 ESAW系統硬體設計 52 圖2-2 (A) ESAW系統硬體設計示意圖 (B)改裝一套市售的高頻震盪電路板的方式 55 圖2-3 315 MHz表面聲波共振元件 57 圖2-4 表面聲波共振元件的內部結構 58 圖2-5 三種電極的照片 61 圖2-6 去除電極外殼的方法 62 圖2-7 比較剪去SAW元件A腳的干擾效應 65 圖2-8 評估外界環境的阻抗變化 66 圖2-9 研究SAW元件之A接腳與地線間的導線的長度 裝置圖 67 圖2-10 頻率訊號干擾設計 69 圖2-11 ESAW系統的組裝過程的照片 70 圖2-12 金屬銅殼底部及高頻震盪電路板之間的導線長度 效應 73 圖2-13 高頻震盪電路板上的電源電壓效應 75 圖2-14 平板式銅箔電極面積的對靈敏度的效應 77 圖2-15 兩片平板式銅箔電極間距的對靈敏度的效應 79 圖2-16 電極的製作 81 圖2-17 不同安排電極的方式所造成的鄰近電磁場之電力線圖形 83 圖2-18 指叉型電極間隔距離與電磁波電力線穿透表層距離的關係 83 圖2-19 三種電極用來偵測各種濃度之Cu(NO3)2 溶液的訊號 84 圖2-20 ESAW系統在溶液中的溫度效應 86 圖2-21 溶液的介電性質效應 88 圖2-22 S-型偵測電極之水溶性醇類效應 89 圖2-23 Q-型偵測電極偵測各種濃度之乙醇和甘油溶液的 訊號 90 圗3-1 ESAW系統偵測鹼金族與銨根之硝酸鹽水溶液的訊號變化 96 圗3-2 ESAW系統偵測鹼土族之硝酸鹽水溶液的訊號變化 97 圗3-3 ESAW系統偵測一些過渡金屬之硝酸鹽水溶液的訊號變化 98 圗3-4 ESAW系統偵測一些鉀鹽水溶液的濃度效應 100 圗3-5 ESAW系統以C-型電極測量弱酸—弱鹼滴定。 103 圗3-6 ESAW系統C-型電極兩電極間的填充物對於頻率 訊號的影響 105 圗3-7 (ΔF/ C )值對(μ×q)的作圖 108 圗3-8 (A)水分子形成排列狀態與恢復散亂狀態的示意圖。 (B) ESAW系統中電極的315 MHz電磁波脈衝示意圖 112 圖4-1 ESAW系統之C/Ag型電極在過氧化氫水溶液中 的頻率訊號 118 圗4-2 ESAW系統C/Ag型電極兩電極間的填充物對於 過氧化氫水溶液頻率訊號的影響 119 圗4-3 不同醣類對GOD催化H2O2分解反應的影響 122 圖4-4 H2O2濃度對GOD催化H2O2分解反應延遲時間的 影響 124 圖4-5 葡萄糖濃度對GOD催化H2O2分解反應延遲時間 的影響 125 圗4-6 ESAW系統在1 mM葡萄糖水溶液中偵測的頻率 訊號 128 圗4-7 ESAW系統在葡萄糖與半乳糖水溶液中偵測的頻率訊號 129 圗4-8 ESAW系統搭配葡萄糖氧化酶所測得的葡萄糖濃度效應 131 圗4-9 ESAW生化感測器偵測葡萄糖氧化酶的濃度效應 132 圖5-1 QCM偵測系統硬體設計之示意圖 138 圖5-2 裝設QCM偵測系統的基本流程 139 圖5-3 實驗裝置中放置QCM電極及待測溶液的方法 140 圖5-4 製作C60-膽固醇水解酶固定化酵素的裝置 144 圖5-5 QCM偵測系統在油酸/非離子型界面活性劑NS 乳化液中的頻率訊號 147 圖5-6 油酸乳化液頻率訊號變化的非離子型界面活性劑 濃度效應 148 圖5-7 石英壓電晶片電極之C60-Cryptand-22塗佈效應 151 圖5-8 油酸的頻率感應檢量線 153 圖5-9 非離子型界面活性劑的對照組空白實驗 155 圖5-10 非離子型界面活性劑與膽固醇混合物的對照組空白實驗 156 圖5-11 非離子型界面活性劑與膽固醇水解酶混合物的對照組空白實驗 157 圖5-12 QCM偵測系統在膽固醇水解酶催化下的頻率訊號 159 圖5-13 油酸膽固醇酯的檢量線 162 圖5-14 偵測油酸膽固醇酯水解後的訊號與直接測定油酸的訊號比較 163 圖5-15 QCM電極塗佈C60吸附膽固醇水解酵素的頻率訊號 165 圖5-16 QCM偵測系統在膽固醇水解酶催化下的頻率訊號 167 圖5-17 C60固定化膽固醇水解酶濾片的活性期 168 圖5-18 油酸膽固醇酯的檢量線(固定化酵素) 169 表1-1 碳六十在不同溶劑中溶解度 2 表1-2 電化學與光學檢測法之比較 14 表1-3 常用之壓電基材的物理性質 26 表1-4 各種電磁波輻射所構成的雜訊來源 37 表2-1 所用C-型電極規格 76 表3-1 金屬陽離子的半徑 95 表3-2 陰離子的半徑 99 表3-3 ESAW 系統在離子水溶液中的偵測極限 102 表3-4 離子在無限稀釋水溶液中的移動率 106 表3-5 一些離子的性質 107 表3-6 KNO3水溶液濃度10-3 ~10-2 M範圍內的訊號變化 110

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