建立PPP-in-tube SPE-ICP-MS連線分析系統進行活體動物肝臟細胞間液中奈米銀粒子的連續動態監測分析研究

Online In-tube Solid Phase Extraction Coupled with Push-pull Perfusion Sampling and ICP-MS Determination for in vivo Monitoring Extracellular Silver anoparticles in Rat Liver

黃靖雯

指導教授 ： 孫毓璋

國立清華大學/原子科學院/生醫工程與環境科學系/碩士(2010年)

摘要

近幾年來，隨著奈米科技的蓬勃發展，奈米科技相關產品已廣泛地進入我們的生活周遭。由於奈米銀粒子具有相當優異的抗菌效果，所以在市面上奈米相關的商品中，從創傷敷料、襪子，甚至到食物保鮮盒等，都可以發現含奈米銀粒子的產品；含奈米銀粒子的產品亦已成為目前奈米相關產品當中佔有率最大者，其市佔率約為50%。然而，雖然奈米銀粒子已廣泛地被應用於生活當中，但因為受限於分析技術，目前文獻中有關動物體內奈米銀粒子的毒理動力學與風險評估的資訊仍相當缺乏。
為了獲得活體動物體內奈米銀粒子濃度的動態變化資訊，本研究建立了一套操作簡單的自動化連線分析系統，該系統中包含了活體抽壓導管採樣技術、奈米銀粒子管內固相萃取技術以及感應耦合電漿質譜儀的偵測技術。在動物實驗過程當中，本研究係由肝臟中採集奈米銀粒子，經由PTFE管進行萃取以達到與複雜生物基質分離之目的，最終則係以ICP-MS進行連續的偵測。
本研究在完成系統的最適化參數探討後，即根據最適化之條件進行連線系統的分析效能測試；實驗結果顯示，本研究所建立之連線分析系統之偵測極限可達0.64 μg/L。在分析可信度確認方面，本研究係將奈米銀粒子添加於模擬生物體液樣品中進行分析；結果顯示，利用所建立之分析技術確實可準確地測定含高濃度生物基質樣品中奈米銀粒子的濃度。此外，根據本研究實際進行大鼠肝臟細胞外液間奈米銀粒子濃度連續監測的結果顯示，本研究所建立之PPP-in-tube SPE-ICP-MS確實具有長時間監測活體動物體內奈米銀粒子動態變化趨勢的可行性。

關鍵字

感應耦合電漿質譜儀 ； 抽壓導管 ； 固相萃取 ； 奈米銀粒子 ； 活體

並列摘要

Recently, the massive progression of nanotechnology has led a large number of applications and consumer goods. Around 50% of nanotechnology-based consumer products were made of silver nanoparticles (AgNPs), making them applicable to wound dressings, socks, and food containers due to the effective inhibition of growth in various microorganisms. Being applied for human use, however, the biokinetic and risk of AgNPs in mammal were rarely assessed, because there is still lack of an adequate analytical technique can provide dynamic information of AgNPs in mammal up to now.
To in vivo monitoring of AgNPs in the extracellular fluids of rat liver, we developed an unsophisticated, automatic, and online hyphenated system comprising push-pull perfusion (PPP) sampling, the established in-tube solid phase extraction (SPE) procedure, and inductively coupled plasma mass spectrometry (ICP-MS) in this study. This system takes advantage of adsorbing AgNPs onto the inner wall of polytetrafluoroethylene (PTFE) tubing which means AgNPs can be extracted from the complicated biological matrix. To optimize the analytical performance, the effect of sampling flow rate, sample loading flow rate, sample pH, tubing inner diameter and blood matrix were investigated. It has been demonstrated that under the optimized conditions, the detection limit of analyte AgNPs were found in the range of sub–g/L. The accuracy of our proposed system was confirmed by analyzing biological samples spiking with defined amounts of AgNPs to demonstrate its validity. After the validation of this method, the applicability of our developed system was further demonstrated by in vivo monitoring the dynamic variation in the concentrations of AgNPs in the liver of anesthetized rats after intravenous dosing.

並列關鍵字

ICP-MS ； push-pull perfusion ； solid phase extraction ； silver nanoparticles ； in vivo

參考文獻

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