鐵是維持生命的重要微量元素之一，分析及測定生物體及環境中的鐵物種，對於提升人類健康與生活品質是不可缺乏的重要一環，由於濃度低與易受鹽類基質干擾等因素的限制，發展高效率、低污染風險且操作簡單的樣品前處理系統，對於了解生物醫學與環境科學領域中鐵物種相關研究而言，至今仍是一個十分重要的課題；為解決上述難題，本研究利用PVC粉填充微管柱連結微透析取樣裝置，搭配閥相輔助注入分析系統與感應耦合電漿質譜儀，發展自動化MD-PVC-packed dual minicolumn-ICP-MS連線分析系統，用來監測活體大鼠腦中，因急性去極化效應，所造成腦細胞間液內Fe(II)與Fe(III)的即時動態變化趨勢，根據所獲得之結果說明，當受到急性去極化刺激之後，腦細胞間液內Fe(II)與Fe(III)的濃度確實會發生顯著降低的情形，亦證實此分析系統確實具有監測活體動物腦中鐵物種動態變化趨勢的能力；此外，本研究亦利用三維列印微管柱固相萃取裝置串聯閥相輔助注入分析系統與感應耦合電漿質譜儀，建立自動化3DP-dual minicolumn-ICP-MS連線分析系統，用來分析環境水樣中的鐵物種，根據分析參考樣品（1643e、1640a、SLEW-3與CASS-4）與添加分析試驗之結果，說明本研究所開發之自動化連線分析系統，確實具有準確分析環境水樣中低濃度鐵物種的能力。

Exploration of brain extracellular non-protein-bound/diffusible iron species remains an important issue in investigations of free radical biology and neurodegenerative diseases. In this study, a sample pretreatment scheme, involving poly(vinyl chloride)–packed minicolumn as a selective extraction device, was optimized in conjunction with microdialysis sampling and inductively coupled plasma mass spectrometry (ICP-MS) in cool-plasma mode for in vivo online monitoring of rat brain extracellular Fe(II) and Fe(III). After system’s optimization, the method’s applicability was verified through (i) spike analyses of offline-collected rat brain microdialysates, (ii) determination of the basal Fe(II) and Fe(III) concentrations of living rat brain extracellular fluids, and (iii) monitoring of the dynamic changes in the Fe(II) and Fe(III) concentrations in response to perfusion of a high-K+ medium.

Furthermore, to enable speciation of trace iron in environmental samples, a stereolithographic 3D printer and acrylate-based resins were utilized to fabricate a demountable minicolunm to selectively extract Fe(II) and Fe(III) and facilitate their analyses in high-salt-content environmental samples by hyphenating to ICP-MS. After system’s optimization, the analyses of the reference materials 1643e, 1640a, CASS-4, and SLEW-3 as well as the spike analyses of those collected environmental samples were performed to confirm the method’s reliability. These two proposed sample pretreatment schemes, based on respectively using the PVC-packed minicolumn and the non-functionalization 3D-printed minicolumn, appear to have great practicality for the iron speciation in biological and environmental samples.

摘要I
Abstract II
目錄III
圖目錄VIII
表目錄XI
第一章前言1
1.1 腦組織中的鐵離子1
1.1.1 鐵離子在腦組織中扮演的角色1
1.1.2 鐵離子對於腦部慢性疾病的影響3
1.2 環境水體中的鐵離子5
1.2.1 鐵離子在環境中扮演的角色5
1.2.2 鐵離子對於環境生態的影響7
1.3 鐵物種分析方法與限制9
1.3.1 鐵物種分析方法之演進9
1.3.2 鐵離子樣品在偵測時所面臨的困境10
1.4 線上前處理分析系統12
1.4.1 線上前處理分析系統之優點12
1.4.2 三維列印技術之應用14
1.5 研究目的16
第二章 儀器分析及原理17
2.1 微透析取樣法17
2.2 感應耦合電漿質譜法18
2.2.1 樣品導入系統20
2.2.2 游離源23
2.2.3 真空緩衝介面與離子聚焦透鏡25
2.2.4 四極柱質量分析器28
2.2.5 離子偵測器29
2.3 三維列印技術31
第三章 實驗材料與方法36
3.1 儀器裝置與配件36
3.1.1 儀器裝置與配件36
3.1.2 實驗藥品與試劑38
3.1.3 實驗用水與容器清洗38
3.1.4 實驗動物來源39
3.2 分析系統的清洗與保存39
3.2.1 微透析探針於實驗前後的清洗與保存39
3.2.2 實驗前後，固相萃取系統的清洗與保存40
3.3 MD-PVC-packed minicolumn-ICP-MS連線分析系統建立與最佳化條件40
3.3.1 MD-PVC-packed minicolumn-ICP-MS連線分析系統建立 40
3.3.2 PVC微管柱製備45
3.3.3 MD-PVC-packed minicolumn-ICP-MS連線分析系統最佳化之探討 45
3.3.4 MD-PVC-packed minicolumn-ICP-MS連線分析系統效能評估 46 3.3.5 MD-PVC-packed minicolumn-ICP-MS連線分析系統應用於監測活體大鼠腦中細胞間液內鐵離子的動態分析48
3.4 3DP-minicolumn-ICP-MS連線分析系統49
3.4.1 三維列印微管柱的設計與列印49
3.4.2 3DP-minicolumn-ICP-MS連線分析系統建立52
3.4.3 3DP-minicolumn-ICP-MS連線分析系統最佳化探討57
3.4.4 3DP-minicolumn-ICP-MS連線分析系統效能評估58
3.4.5 樣品取得與配置59
第四章 實驗結果與討論61
4.1 MD-PVC-packed minicolumn-ICP-MS連線分析系統之最佳化條件探討61
4.1.1微透析探針取樣的回收率62
4.1.2樣品pH值對於萃取效率的影響63
4.1.3緩衝溶液濃度對於萃取效率的影響64
4.1.4載流溶液體積對於分離效率的影響65
4.1.5載流溶液流速對於萃取效率的影響66
4.2 MD-PVC-packed minicolumn-ICP-MS連線分析系統效能評估67 4.2.1區分曲線的建立69
4.2.2檢量線與系統效能評估69
4.2.3分析系統之長時間穩定度評估70
4.3利用MD-PVC-packed minicolumn-ICP-MS連線系統進行活體大鼠腦中細胞間液內鐵離子的動態分析71
4.4 3DP-minicolumn-ICP-MS連線分析系統之最佳化探討74
4.4.1樣品pH值對於萃取效率的影響74
4.4.2樣品進樣流速對於萃取效率的影響76
4.4.3緩衝溶液濃度對於萃取效率的影響77
4.4.4載流溶液體積對於分離效率的影響78
4.4.5載流溶液流速對於分離效率的影響79
4.4.6三維列印微管柱內部結構之最佳化探討80
4.4.7三維列印微管柱飽和吸附量81
4.5 3DP-minicolumn-ICP-MS連線分析系統效能評估82
4.5.1 線性關係與偵測極限83
4.5.2系統準確度及精密度評估83
4.6 利用3D-printed-minicolumn-ICP-MS連線分析系統進行環境水樣中鐵物種之分析研究86
第五章 結論88
第六章 參考文獻90
附錄99

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