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研究生: 周秉毅
Jhou, Bing-Yi
論文名稱: 利用分子模印技術搭配液相層析串聯式質譜儀分析IQ及IQx型的雜環胺類
Analysis of the IQ- and IQx-type Heterocyclic Amines by Molecularly Imprinted Technology and LC-MS/MS
指導教授: 陳頌方
Chen, Sung-Fang
學位類別: 碩士
Master
系所名稱: 化學系
Department of Chemistry
論文出版年: 2018
畢業學年度: 106
語文別: 中文
論文頁數: 100
中文關鍵詞: 雜環胺化合物分子模印聚合物2-丙烯醯胺基-2-甲基丙磺酸甲基丙烯酸液相層析-串聯式質譜儀固相萃取技術
英文關鍵詞: Heterocyclic amines, Molecularly imprinted polymers, AMPS, MAA, LC-MS/MS, Solid-phase extraction
DOI URL: http://doi.org/10.6345/THE.NTNU.DC.042.2018.B05
論文種類: 學術論文
相關次數: 點閱:139下載:2
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  • 雜環胺化合物是指在其化學結構中含有兩種以上不同元素和至少一個胺基(-NRR')的環狀結構化合物。許多文獻和臨床研究已指出雜環胺化合物具有高度致癌性和致突變性。分子模印技術是基於“抗原-抗體結合理論”的延伸性應用,該技術提供對目標分析物極佳的特異性和識別能力,而利用該技術合成的高分子材料則稱為分子模印聚合物(MIPs)。本篇研究目的為開發具有高度選擇性的分子模印聚合物,並使用液相層析-串聯式質譜技術定量複雜基質中的雜環胺化合物。本研究選擇IQ作為模板分子,並使用兩種官能基單體AMPS和MAA,以及交聯劑EGDMA分別透過總體聚合和沉澱聚合的方式合成分子模印聚合物,並搭配使用掃描式電子顯微鏡觀察兩種分子模印聚合物的表面特徵。之後透過對模板分子IQ的吸附能力來評估與優化合成比例,結果顯示當模板分子:官能基單體:交聯劑的比例為1:8:40時具有最好的吸附效果。此外,我們將兩種分子模印聚合物與固相萃取法結合並優化其萃取條件。實驗結果為使用MAA作為官能基單體所製備的分子模印聚合物不僅對 IQ具有高度選擇性,且MeIQ和8-MeIQx兩個雜環胺也可以被有效地偵測。最後,本實驗開發的方法可以成功應用於真實樣品的分析。

    Heterocyclic amines (HCAs) refer to compounds that have ring structure which contains more than two different elements and at least one amine group (-NRR') in their chemical structures. Many studies and literature have pointed out that HCAs are highly carcinogenic and mutagenic. Molecularly imprinted technology is an extending application based on the "antigen-antibody binding theory". This technology provides excellent specificity and recognition ability, and the material synthesized by this method called molecularly imprinted polymers (MIPs).  The aim of this study was to develop the highly specific molecularly imprinted polymers for the purification and quantification of heterocyclic amine compounds in complex matrices using liquid chromatography-tandem mass spectrometry. In this study, IQ was selected as a template molecule, and two functional monomers, AMPS and MAA were used with cross-linking agent EGDMA for the synthesis of molecularly imprinted polymers by means of bulk polymerization and precipitation polymerization. The characteristics and surface of the MIPs were investigated through the scanning electron microscope. Afterwards, the synthesis ratios were evaluated and optimized by their adsorption capacity to the template molecules and the selectivity to other HCAs. As a result, template/functional monomer/cross-linking agent at a ratio of 1:8:40 gave better adsorption effect. Furthermore, two kinds of molecularly imprinted polymers were incorporated with solid-phase extraction and optimized the extraction conditions. The results indicated that excellent selectivity for IQ was obtained using MAA as functional monomer, while MeIQ and 8-MeIQx could also be effectively detected. Finally, our method was successfully applied to the real samples.

    謝誌 I 目錄 II 圖目錄 VI 表目錄 VIII 中文摘要 IX Abstract X 縮寫 XI 第一章 序論 1 第一節 研究動機 1 第二節 雜環胺化合物 4 壹、 雜環胺化合物的簡介 4 貳、 食品中的雜環胺化合物 4 參、 雜環胺化合物的致癌性 7 第三節 分子模印技術 9 壹、 分子模印技術的簡介 9 貳、 分子模印技術的發展及應用 10 參、 分子模印聚合物的組成 11 肆、 分子模印聚合物的合成技術 14 第四節 樣品前處理 16 壹、 固相萃取技術(Solid-Phase Extraction, SPE) 16 貳、 分子模印技術搭載固相萃取法(MISPE) 17 第五節 高效能液相層析系統 18 壹、 層析原理 19 貳、 吸附層析法(Adsorption Chromatography) 20 第六節 質譜分析系統 21 壹、 進樣系統(Sample Inlet) 22 貳、 離子源(Ion Source) 23 參、 質量分析器(Mass Analyzer) 27 肆、 偵測器(Detector) 33 第二章 實驗方法與器材 34 第一節 實驗設備 34 第二節 以AMPS為官能基單體製備分子模印聚合物 35 壹、 材料與試劑 35 貳、 實驗步驟 35 第三節 AMPS-MIPs模板分子清洗 37 壹、 材料與試劑 37 貳、 實驗步驟 37 第四節 AMPS-MIPs/NIPs的等溫吸附 38 壹、 材料與試劑 38 貳、 實驗步驟 38 第五節 AMPS-MIPs/NIPs的動力吸附 40 壹、 材料與試劑 40 貳、 實驗步驟 40 第六節 以MAA為官能基單體製備分子模印聚合物 41 壹、 材料與試劑 41 貳、 實驗步驟 41 第七節 MAA-MIPs模板分子清洗 43 壹、 材料與試劑 43 貳、 實驗步驟 43 第八節 MAA-MIPs/NIPs的等溫吸附 44 壹、 材料與試劑 44 貳、 實驗步驟 44 第九節 MAA-MIPs/NIPs的動力吸附 46 壹、 材料與試劑 46 貳、 實驗步驟 46 第十節 分子模印技術搭載離心式固相萃取法 47 壹、 材料與試劑 47 貳、 實驗步驟 47 第十一節 分子模印聚合物對雜環胺的選擇性吸附 49 壹、 材料與試劑 49 貳、 實驗步驟 49 第十二節 液相萃取法製作空白基質樣品 51 壹、 材料與試劑 51 貳、 實驗步驟 51 第十三節 掃描式電子顯微鏡分析 52 壹、 掃描式電子顯微鏡設備 52 貳、 實驗步驟 52 第十四節 模板清洗溶液的分析 53 壹、 紫外光-可見光吸收光譜儀設備 53 貳、 實驗步驟 53 第十五節 LC-MS/MS分析 54 壹、 高效能液相層析設備 54 貳、 高效能液相層析參數設定 54 參、 串聯式質譜儀設備 55 肆、 串聯式質譜儀參數設定 55 伍、 數據分析軟體 55 第三章 結果與討論 56 第一節 高效能液相層析儀之參數優化 56 壹、 層析管柱的選擇 56 貳、 移動相的選擇與梯度的優化 57 第二節 串聯式質譜儀之參數優化 59 壹、 游離源參數設定 59 貳、 離子對的選擇與優化 60 第三節 分子模印聚合物的合成條件優化 61 壹、 致孔溶劑的選擇與優化 62 貳、 合成比例的優化 68 第四節 分子模印聚合物的清洗 70 第五節 分子模印聚合物的等溫吸附 72 壹、 兩種分子模印聚合物的等溫吸附討論 72 貳、 以數學模型評估分子模印聚合物之特性 75 第六節 分子模印聚合物的動力吸附 78 第七節 分子模印聚合物的選擇性吸附 81 第八節 分子模印聚合物搭載離心式固相萃取之優化 82 第九節 方法確校 84 壹、 檢量線(Calibration curve)的製作 84 貳、 準確度(Accuracy)及精密度(Precision) 86 參、 基質效應(Matrix effect) 88 第四章 結論與未來展望 90 第五章 參考文獻 91

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