毛細管電泳技術的方法開發與應用

Development and Application of Capillary Electrophoresis Techniques

邱貞文(Chen-Wen Chiu)

指導教授 ： 黃悉雅

中原大學/理學院/化學研究所/博士(2006年)

https://doi.org/10.6840/cycu200600109

摘要

隨著時代的進步科技的發展，分析技術因應研究需要而必須不斷地創新，以往在分析技術上的開發，常常只侷限於標準品的測試，而非應用於基質干擾較大且純化步驟繁瑣的實際樣品上。因此，本研究主要目的為發展毛細管電泳技術在實際樣品分析上的應用，並且將對所應用的三種電泳分離模式作一系列深入的探討。
第一部份為以毛細管電泳結合大體積樣品堆積技術分析食品中所含食用色素。實驗中將探討樣品堆積時間、注射時間以及樣品基質等因素對分離效率的影響。由實驗結果顯示，以毛細管電泳添加環糊精分析食用色素效果極佳，且利用大體積樣品堆積技術可增加偵測靈敏度，最後我們以最佳化條件配合固相萃取法成功地應用於分析牛奶、汽水、果凍等實際樣品中所含色素。
第二部份為以毛細管電層析技術分析食品、藥品及化妝品中所含防腐劑與防曬劑。實驗中將探討聚合條件變化對甲基丙烯酸酯高分子靜相之孔徑、形態、熱性質及分離效率的影響。由實驗結果可知，隨著孔洞溶劑中正丙醇比例增加，高分子靜相孔徑減小，表面積增加；而單體中AMPS比例增加時會造成孔徑變大，提供更多多表面積，增加分離效率。另一方面，由研究中指出動相pH值、離子濃度、乙腈比例、樣品基質與分離電壓等因素對分離效率也具有相當程度的影響。最後，我們將利用簡單的前處理方法以最佳化條件成功地應用於實際樣品分析上。
第三部份為以非水相毛細管電泳系統連接電灑質譜儀分析五種常見之菸鹼。實驗中將探討不同背景電解液(醋酸銨、甲酸銨)對分離效率與質譜靈敏度的影響，其中，pH值、離子濃度及有機溶劑比例為其變化因素。尋找出最佳化分離條件後將直接應用於質譜檢測上，並調整鞘流組成及流速、霧化氣體流速、乾燥氣體流速及溫度等質譜參數直至菸鹼在質譜中得到最佳感度。最後，我們以市售香菸作為實際樣品，檢測所含之菸鹼-尼古丁。

關鍵字

毛細管電泳 ； 電灑-質譜 ； 毛細管電層析 ； 大體積樣品堆積

並列摘要

Analytical techniques have been well developed and progressed apparently in the recent years. However, the analyses of real sample were always limited because of its complicated matrix. In order to improve their separation ability on real sample, several methods based on CE were developed in the study.
In the first part of this study, a large-volume sample stacking (LVSS) technique that combined with a high-efficiency CE was developed to analyze eight food colorants in food samples. The effects of several factors, such as sample stacking time, sample injection length and sample matrix, on the stacking ability were discussed. In addition, the usage of LVSS on colorant detection was able to reduce the detection limits successfully in CE method. The proposed LVSS-CE method was applied to determine the content of the colorant additives in commercial foods without matrix interference.
In the second part, the analyses of six preservatives and eight benzophenones by capillary electrochromatography (CEC) using methacrylate ester-based monolithic columns were developed. The properties of the polymeric column were affected significantly by the preparation conditions of polymeric monolithic column. Several factors including the ratio of AMPS to monomer and the composition of porogenic solvent were examined in order to achieve optimal CEC separation. Moreover, the morphology, porosity, thermal stability and surface area of polymeric columns were characterized by scanning electron microscope (SEM), mercury intrusion porosimeter (MIP), thermogravimetry (TGA) and Brunauer Emmett Teller method (BET). The ratio of AMPS to monomer determined the amount of sulfonic group on the monolithic surface, and also influenced the magnitudes of electroosmotic flow (EOF) in the monolithic column. In addition, the porosity of monolithic polymer could be altered by the ratio of monomer to porogenic solvent. Finally, the feasibility of CEC on the analyses of preservatives and benzophenones in commercial products was also discussed.
In the third part, the separation of tobacco alkaloids by nonaqueous capillary electrophoresis (NACE) combined with electrospray ionization/mass spectrometer (ESI/MS). First, the effects of types of background electrolyte, buffer pH, ion strength and organic solvent level on CE separation were examined. After that, various MS parameter including the composition and flow rate of sheath liquid, flow rate of nebulizing gas and dry gas, and temperature of ESI on the mass sensitivity of tobacco alkaloids were optimized. Finally, the content of tobacco alkaloids in commercial tobacco products was determined by the NACE/ESI/MS method.

並列關鍵字

CEC ； CE ； ESI-MS ； LVSS

參考文獻

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