本研究主要分為兩個部分。第一部分是利用電激發化學發光法( ECL, Electrogenerated chemiluminescence )搭載自動化流注系統( flow injection analysis, FIA )偵測糖尿病用藥米格列醇，主要是利用電化學的方式使Ru(bpy)32+在電極表面氧化產生Ru(bpy)33+並催化三級胺類米格列醇使米格列醇形成自由基，米格列醇自由基再與Ru(bpy)33+反應形成激發態Ru(bpy)32+＊而其由激發態返回基態時放出冷光(λ=620nm)以偵測冷光的方式達到定量米格列醇的目的，此FIA-ECL偵測系統在最佳化的條件下線性範圍為0.1 μM ~ 10 μM (R2=0.999)，偵測極限(S/N=3)為55.3 nM，更大的線性範圍可達15 μM (R2=0.998)，偵測極限(S/N=3)為58.11 nM，相對標準偏差(RSD)為0.52%。第二部分則是將此系統應用於高效液相層析儀( HPLC )上，將干擾物與米格列醇分離以達到在尿液生化樣品上分析應用的目的其線性範圍為1 μM~30 μM，偵測極限(S/N=3)為0.78 μM，此分析特性也符合在臨床上檢測之使用。

There are two parts in this study. The first one, determination of miglitol with Electrogenerated chemiluminescence ( ECL ) with flow injection analysis ( FIA ) was fabricated. In electrode surface Ru(bpy)32+ go through electron-transfer reaction with miglitol (co-reactant) to form excited state Ru(bpy)32+＊, and excited state Ru(bpy)32+＊ back to the ground state that emit light by vibrational energy relaxation( VR ). In this analysis system, the linear range of miglitol is between 0.1 μM and 10 μM (R2=0.999), sensitivity is 570.5 /μM, detection limit (S/N=3) is 58.11 nM, and relative standard deviation of twenty repetitive detection is 0.52%. 
The second one, we combine ECL with HPLC to seprate interference in urine samples. HPLC is used to separate miglitol and matrix in urine samples and determination of miglitol by PMT. The linear range in HPLC-ECL system is between 1 μM and 30 μM, detection limit (S/N=3) is 0.78 μM.

總目錄
總目錄	iii
圖目錄	vi
表目錄	viii
第一章 緒論	1
1-1藥物偵測之重要性	1
1-2米格列醇( Miglitol )	2
1-2-1米格列醇的藥理及用途	2
1-2-2米格列醇的偵測方法	6
1-3糖尿病	7
1-3-1糖尿病及其診斷方式	7
1-3-2糖尿病種類與病因	8
1-3-3胰島素	11
1-3-4蛋白質的糖化	12
1-3-5糖尿病的併發症	14
1-2-6糖尿病常見用藥及種類	15
1-4電激發化學發光法	17
1-4-1電激發化學發光法的發展與應用	17
1-4-2電激發化學發光法的原理	19
1-4-3電激發化學發光法的淬滅	29
1-5自動化流注分析系統	30
1-6高效液相層析系統	32
1-7 LabVIEW簡介	40
1-8本研究之目的	42
第二章 實驗步驟	43
2-1儀器	43
2-2藥品	47
2-3實驗步驟與設計	48
2-3-1系統操作電位的探討	48
2-3-2初步緩衝液酸鹼值與增強劑的探討	49
2-3-3緩衝溶液酸鹼值的探討	49
2-3-4緩衝溶液種類的探討	49
2-3-5緩衝溶液濃度的探討	50
2-3-6發光體Ru(bpy)32+濃度的探討	50
2-3-7樣品迴路體積的探討	51
2-3-8光電倍增管操作電壓的探討	51
2-3-9分析特性評估	51
2-3-10電激發化學發光法在HPLC上之應用	52
第三章 結果與討論	53
3-1工作原理的探討	53
3-2最佳化條件的探討	57
3-2-1系統操作電位	57
3-2-2初步緩衝液酸鹼值與增強劑	60
3-2-3緩衝溶液酸鹼值	63
3-2-4 緩衝溶液種類	66
3-2-5緩衝溶液濃度	68
3-2-6發光體Ru(bpy)32+濃度	71
3-2-7樣品迴路體積	73
3-2-8光電倍增管操作電壓	75
3-3分析特性的探討	77
3-4米格列醇電激發化學發光系統於HPLC上之應用	84
3-4-1尿液生化樣品與米格列醇之分離條件	85
3-4-2米格列醇在HPLC-ECL偵測系統上之校正曲線	92
3-5結論	95
參考文獻	96
附錄	104
 
圖目錄
圖 1米格列醇之化學結構	4
圖 2米格列醇在小腸中的作用	5
圖 3糖化作用之反應機制	13
圖 4消滅型電激發化學方光之反應機制	21
圖 5 Oxalate / Ru(bpy)32+系統之ECL反應機制	24
圖 6胺類ECL反應機制1	26
圖 7胺類ECL反應機制2	26
圖 8胺類ECL反應機制3	27
圖 9胺類ECL反應機制4	27
圖 10 TPrA / Ru(bpy)32+系統產生激發態Ru(bpy)32+之途徑	28
圖 11六向注射閥原理	34
圖 12滯留時間關係圖	36
圖 13分析物在分離管柱中之分散	37
圖 14解析度與波峰之關係圖	40
圖 15電激發化學發光系統之儀器設備	44
圖 16放大之樣品反應槽結構	45
圖 17HPLC-ECL系統之儀器設備	46
圖 18 Ru(bpy)32+之化學結構	47
圖 19米格列醇循環伏安圖與電激發化學發光訊號。	55
圖 20米格列醇之可能反應機制。	56
圖 21米格列醇偵測系統之操作電位的探討。	59
圖 22初步緩衝溶液酸鹼值探討。	61
圖 23電激發發光系統之增強劑的探討。	62
圖 24米格列醇偵測系統之緩衝溶液酸鹼值的探討。	65
圖 25米格列醇偵測系統之緩衝溶液種類的探討。	67
圖 26米格列醇偵測系統之磷酸緩衝溶液濃度的探討。	69
圖 27米格列醇偵測系統之硼酸緩衝溶液濃度的探討。	70
圖 28米格列醇偵測系統之發光體Ru(bpy)32+濃度的探討。	72
圖 29米格列醇偵測系統之樣品迴路體積的探討。	74
圖 30米格列醇偵測系統之PMT操作電壓的探討。	76
圖 31米格列醇偵測系統之校正曲線。	79
圖 32 PMT操作電壓對校正曲線之影響	80
圖 33米格列醇偵測系統之再現性。	81
圖 34米格列醇偵測系統之干擾物探討。	83
圖 35米格列醇在HPLC-ECL系統上酸鹼值與訊號強度的探討。	87
圖 36米格列醇在HPLC-ECL系統上酸鹼值與解析度的探討。	88
圖 37 米格列醇在HPLC-ECL偵測系統上酸鹼值之真實訊號。	91
圖 38尿液生化樣品中干擾物之真實訊號	93
圖 39米格列醇濃度對滯留時間之影響。	94

表目錄
表 1糖尿病之診斷方法	8
表 2抗糖尿病藥物種類及機制	16
表 3米格列醇之分析特性表	82

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