臺灣博碩士論文加值系統

糖尿病是全世界先進國家揮之不去的夢魘，絕大多數的糖尿病是由肥胖所導致的，而糖尿病患可以藉由血糖濃度的監控而獲得病情的改善，甚至痊癒，於是一套便宜又能即時監測的血糖感測器正是糖尿病患者所需要的。
本研究希望可以建立一操作簡單而價格便宜的化學放光感測器，並且將其運用在偵測葡萄糖/過氧化氫的濃度偵測。本研究利用矽偵測器搭配上放大器當作化學放光的量測系統，自製生醫晶片，並以蠕動幫浦以及HPLC之高壓幫浦推送，建立流道與晶片兩種系統，進行化學放光實驗的量測。
本實驗結果顯示，流道系統的可以找出濃度與放光強度的線性相關，在晶片系統可以找出濃度與放光強度的相對關係；而在葡萄糖的部分則是探討其未能有效量測發光效果的原因。

Abstract
Diabetes mellitus (DM) is a hot issue for developed countries on earth, and most of the diabetics are due to obese. DM can be controlled by monitoring the change of the amount of blood sugar real time, even be fully recover. Hence, a system of cheaper, easier operating, monitoring real time is very in want of diabetics.
In this study, we want to establish an optical system of cheaper, easilier operating, monitoring real time, and use it to detect the concentration of glucose/ hydrogen peroxide. We use a silicon detector with a amplifier in which to enhance signals, and a biosensor, and obtained power from peristalsis pump and HPLC pump. We established two fluid system (fluid injection system and biosensor system) to detect the intensity of chemiluminescence (CL) from the chemical reaction.
Our experiment shows that the fluid injection system can find the linear correlation between sample concentration and chemiluminescence signal intensity, and we can also find the correlation between sample concentration and signal intensity in sensor system. Finally, we discuss the reason that why chemiluminescence did not react when we injected glucose, can try to decide them.

第一章 緒論………………………………………………………..1
1.1. 糖尿病簡介………………………………………………….1
1.2. 糖尿病所造成的危害……………………….………………3
1.3. 生物晶片……………………………….……………………5
1.4. 研究目的…………………………………….………………6

第二章 原理……………………………………………..…………8
2.1. 化學冷光的介紹與原理…………………………………………8
2.1.1. 化學放光與基本條件………………………………….………8
2.1.2 化學放光電子轉移機制……………………..………………9
2.1.3 化學放光強度…………………………………………………12
2.1.4 常用化學放光材料……………………………………………13
2.2 Luminol與醣類簡介………………………………….…………13
2.2.1 Luminol……………………………………………...………13
2.2.2 葡萄糖…………………………………………………………17
2.3 葡萄糖反應的基本原理…………………………………………18
2.4 酵素固定…………………………………………...…………20
2.4.1 酵素固定法之分類……………………………………………20
2.4.2 固定化酵素的優缺點…………………………………………22
2.4.3 本研究所採用的固定法………………………………………23
2.5化學動力學公式分析……………………………………………23

第三章 實驗方法…………………………………………………26
3.1 葡萄糖濃度偵測系統設計圖………………………..…………26
3.1.1 Luminol - HB2BOB2B - HRP流道系統設計…………………26
3.1.2 Luminol - HB2BOB2B – GOD - HRP晶片系統設計……..27
3.2 實驗流程……………………………….…………………..…28
3.3 藥品………………………………….…………………..……29
3.4 溶液配製…………………………….………………..………29
3.5 儀器裝置……………………………….……………..…………32
3.5.1 光學量測裝置…………………………………………………32
3.5.2 流動注入分析系統 --- 流道系統……………………………34
3.5.3 流動注入分析系統 --- 晶片系統…………………………36
3.6 實驗設計……………………………………………………...…38
3.6.1 矽偵測器放大器製作…………………………………………38
3.6.2 酵素固定化流程 ………………………..…………………40
3.6.2.1 酸洗 …………………………………………...………40
3.6.2.2 矽烷化 …………………………………...………………40
3.6.2.3 酵素固定化………………………………….……………41
3.6.3 晶片置備流程………………………………………………42

第四章 結果與討論………………………………………………47
4.1 自製放大器……………………………………...……………47
4.1.1 設計原理 ………………………………….…..…………47
4.1.2 數種測量儀器之比較…………………………………………48
4.1.3 放大倍率之選擇……………………………………..…..50
4.2 Luminol - HB2BOB2B - HRP流道系統……………..………50
4.2.1 實驗流程…………………………………………………50
4.2.2 流道組參數最佳化設定……………………………..……. 53
4.2.2.1 HB2BOB2B濃度的影響……………………...……...…53
4.2.2.2 HRP濃度的影響………………………………….………57
4.2.2.3 luminol 濃度的影響……………………….…..………62
4.2.2.4 緩衝溶液pH值的影響……….…………………………66
4.2.2.5 蠕動幫浦流速的影響………………………..……………68
4.2.3 參數濃度線性關係分析……………………………………69
4.2.3.1 尋找參數濃度最合適之反應速率方程式……………….70
4.3 Luminol – Glucose - God - HRP晶片系統………………73
4.3.1 Luminol - HB2BOB2B – HRP反應系統……………………73
4.3.1.1 晶片系統與流道系統的比較…………………...……..73
4.3.1.2 晶片系統中HB2BOB2B濃度對放光強度之影響……...……77
4.3.2 Luminol – Glucose - God – HRP 反應系統……………78
4.4 未能有效發光的問題探討………………………………...79
4.5 未來工作……………………………………………………...84
第五章 結論………………………………………………………86
參考資料…………………………………………………………..87

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