臺灣博碩士論文加值系統

現有之紅外光感測器在量測的選擇性上僅能取決於吸收峰之不同，對於複雜的樣品在測量上無法達到期待的選擇性，為增加紅外光感測之選擇性，本研究提出利用紅外光減弱式全反射法結合酵素專一性反應的原理，建立一套具有高選擇性之葡萄糖感測系統。本系統採用流動注入法將樣品注入含有葡萄糖氧化酵素的反應器，其反應產物再以紅外光進行感測，由於所偵測的產物為葡萄糖酸其量少又具有高極性，無法直接以一般ATR方法量測。故此，ATR感測晶體利用雙層式改質法改質上一層具有高度水穩定性的高極性分子。此改質層不僅能夠隔絕水分子，並且對高極性分子有極佳的吸附效果，因此可以應用至葡萄糖酸之檢測。另外為增加實用性與經濟效益，酵素以共價鍵結固定於反應器，達到重複使用之目的。對量測過程中相關的變因影響並深入討論，以建立最佳條件。結果顯示，以感測層吸附待測樣品的速率僅需2分鐘即可獲得訊號，而吸附之樣品可在10分鐘內完全脫附，感測器可循環重複利用。使用填充式的酵素反應器，pH值為5.1的條件下進行葡萄糖的檢測，在含有蔗糖或果糖的葡萄糖分析樣品內，其影響的結果是有限的，對於濃度範圍在0.1 mM~10 mM葡萄糖的偵測線性可達0.99。

Glucose is an important biological compound. To improve the current methods to determine this compound, an infrared (IR) chemical sensing method was developed in this work. Because glucose exhibits high polarity and the samples containing this compound are usually very complex, sensitivity and selectivity are the major considerations in determination. To heighten the sensitivity in detecting this compound with the IR sensing method, a two-layer modification method was developed, in order to eliminate the problem associated in conventional IR sensing methods. To increase selectivity, glucose oxidase was employed. Through a flow injection system, the reaction product, gluconic acid, was easily detected by the IR sensing system developed in this work. To optimize the detection system, factors such as the response time, the effect of pH in the solution, the influence of flow rate, the efficiency in regeneration of the sensing device, and chemical interferences were investigated. The results indicated that the response time can be shorter than 2. Meanwhile, pH influenced the analytical signals significantly and the most suitable pH was around 5. In the examined range of flow rate, results indicated that flow rate was not significant. By examining the analytical of glucose in a solution containing fructose or sucrose, results indicated that the influences were limited. Using optimal conditions, the linear regression coefficients in standard curve can be higher than 0.99 for a concentration range from 0.1 mM to 10 mM.

中文摘要………………………………………………………..………Ⅰ
英文摘要…………………………………………….………………….Ⅱ
謝誌……………………………………………………………………..Ⅲ
總目錄…………………………………………………………………Ⅳ
圖表目錄………………………………………………………………Ⅶ
第一章 緒論………………………………………………………….1
1 — 1前言…………………………………………………………………1
1 — 2酵素感測器之回顧…………………………………………………2
1 — 3研究動機…………………………………………………………4
第二章 原理與機制………………………………………………….6
2 — 1減弱式全反射原理介紹……………………………………………6
2 — 2紅外光感測表面改質原理………………………………………..12
2 — 2 — 1環糊精介紹……………………………………………………14
2 — 2 — 2改質原理………………………………………………………19
2 — 3酵素化學…………………………………………………………..20
2 — 3 — 1酵素的基本特質………………………………………………20
2 — 3 — 2酵素的活性……………………………………………………23
2 — 3 — 3酵素的固定化…………………………………………25
第三章 實驗部分…………………………………………………...30
3 — 1儀器與藥品………………………………………………………30
3 — 2藥品配製…………………………………………………………..33
3 — 3光學元件之製作…………………………………………………35
3 — 3 — 1感測層的合成…………………………………………………35
3 — 3 — 2酵素反的固定化………………………………………………38
3 — 4光學感測系統之建立……………………………………………41
3 — 5 實驗步驟…………………………………………………………43
第四章 結果與討論………………………………………………45
4 — 1感測器之性質討論………………………………………………..45
4 — 1 — 1高分子覆膜厚度之確定………………………………………45
4 — 1 — 2感測層之合成與鑑定………………………………………49
4 — 2感測器之性質討論……………………………………………….54
4 — 2 — 1改質層之水穩定性……………………………………………54
4 — 2 — 2指標樣品的吸收訊號…………………………………………58
4 — 2 — 3感測層的再生…………………………………………………67
4 — 3檢測條件最佳化…………………………………………………..70
4 — 3 — 1酵素反應器的選擇……………………………………………70
4 — 3 — 2 pH值對檢測的影響………………………………………..73
4 — 3 — 3樣品流通速度的討論…………………………………………77
4 — 4真實樣品的測試…………………………………………………..79
4 — 4 — 1選擇性之探討……………………………………………….79
4 — 4 — 2吸附曲線的建立………………………………………………82
4 — 5感測器長期穩定性的追蹤………………………………………87
4 — 6 酵素反應器效能追蹤……………………………………………87
第五章 結論………………………………………………………90
參考文獻………………………………………………………………..91

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