臺灣博碩士論文加值系統

由於人體中微量元素涉及體內之酵素、荷爾蒙、蛋白質及核酸等的代謝過程，因此它對生物體的生理功能扮演著相當關鍵角色。近年來，隨著生命科學的發展，為了增進對生命現象的瞭解，亟需對生物體內各種重要物質的作用機轉進行深入的探討。由於在研究生物體內物質作用機轉的過程中，微小區域內特定生物物質的濃度及其連續動態的變化資訊是該類研究的重要需求。針對上述分析的需求，在分析方法建立的過程中，小量樣品 (數十個µL)，複雜基質及極低分析物濃度 (£ppb)均是不可避免的困難，亦需仰賴先進分析技術的支援與配合。
微透析法(microdialysis) 係一具有微小區域的連續取樣技術；電熱揮發（electrothermal sample）樣品導入技術則擁有低樣品消耗量(20µL)，去除基質及高樣品傳輸效率等優點，若再配合感應耦合電漿質譜儀(ICP-MS)的高靈敏度，同時多元素及快速分析的特點，將可提供一套適合於微量重金屬元素在微小區域內濃度的連續動態的變化資訊。本研究的目的旨在建立一套適合進行離體（in vitro）及體內（in vivo）分析的Microdialysis-ETV-ICP-MS分析系統。研究中分別針對微透析樣品導入技術，ETV及ICPMS最佳化的操作條件，及樣品基質去除過程中所添加的基質修試劑進行探討，並利用樣品基質與透析液（ringer solution）較為相近的標準海水參考樣品，進行方法可靠性的探討。最後，本研究亦進行模擬生物體離體（in vitro）Ringer透析液樣品中鋅、銅、錳及鉛等元素的偵測，以進一步確認所建立之分析方法在面臨真實生物樣品分析時的可行性。

In physiological studies, it is well known that most of the bioavailability and toxicity of trace elements strongly depend on the concentration. In view of the dialysate can be used to reflect the extracellular concentration of metal ions, in-vivo microdialysis coupled with suitable on-line instrumental analytical technique has been widely employed to investigate the physiological reactions in a specific micro-area of a living organism. Owing to the extremely low concentration of analytes and complex matrix are always encountered in the conjunction of analytical chemistry and biological researches, an on-line sampling and matrix separation technique dealing with high salt contented samples has been studied in the present work.
In this study, a hyphenation technique comprising in-vivo microdialysis and ETV-ICP-MS was developed to monitor the dynamic variation of trace metals in simulated body fluid systems. The sampling system was constructed by a micro-injection pump and 6-way valve equipped with a 20-mL sample loop. The interface between sampling system and ETV-ICP-MS was built up with narrow PTFE tubing where the solution was driven by a peristaltic pump. To solve the salt interference caused by the ringer perfusate and to achieve optimal analytical signal, a mixture of palladium and NH4NO3 was employed as binary chemical modifier in the ETV separation procedure. The effects of pyrolysis and vaporization temperature and the amount of added modifiers on the analyte signal were also explored and optimized. Based on the stability and sensitivity studies, the proposed on-line system coupling microdialysis with ETV-ICP-MS has been proven feasible for the determination of trace elements ( Cu, Zn, Mn, Pb ) in biological samples.

Chapter 1. Introduction
1.1 The important of trace elements for biological system 1
1.2 The needs of the information of trace elements in the development of neuroscience 4
1.3 Microdialysis sampling technique 8
1.4 Trace element analysis techniques 9
1.5 Object 12
Chapter 2. Principle
2.1 Microdialysis 13
2.2 Electrothermal vaporization device (ETV) 16
2.3 ICP-MS 18
2.3.1 Inductively Couple Plasma 19
2.3.2 Ion Extraction 23
2.3.3 Ion Fousing 24
2.3.4 Quadrupole Mass Analyser 25
2.3.5 Ion Detection 26
Chapter 3. Experimental
3.1 Apparatus 29
3.1.1 Microdialysis 29
3.1.2 ETV-ICP-MS 29
3.2 Reagents 32
3.2.1 Perfusate 33
3.2.2 Standard Solution and SRM 34
3.2.3 Chemical Modifier 34
3.3 Analytical Procedure 34
3.3.1 Protocol for the use of Microdialysis Probes 34
3.3.2 On-line Analytical System 35
3.3.3 Quantitative Methods 35
Chapter 4. Results and Discussion
4.1 Proposed Microdialysis-ETV-ICP-MS System 37
4.1.1 Possible Interference Resulted from Saline Matrix 38
4.2 Matrix Modifier 40
4.2.1 Effects of NH4NO3 and NH4H2PO4 as Chemical modifier 43
4.2.2 Effect of Pd as a Chemical Modifier 46
4.3 Optimization of temperature program 48
4.3.1 Effect of pyrolysis temperature 48
4.4 Effects of the amount of matrix modifiers 50
4.4.1 Effect of the amount of NH4NO3 50
4.4.2 Effect of the amount of Pd 52
4.4.3 Effect of pyrolysis hold time 54
4.5 Effect of Vaporization Temperature 56
4.6 Analytical performance of direct ETV-ICP-MS method 59
4.7 Analytical performance of microdialysis-ETV-ICP-MS 61
4.7.1 Stability of microdialysis-ETV-ICP-MS 62
4.7.2 Analysis of Ringer solution by on-line microdialysis-ETV-ICP-MS system 63
4.7.3 Concentration Discrimination Capability 64
Chapter 5. Conclusion 68
Chapter 6. Reference 70

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