- 學位論文
毛細管電泳之方法開發與應用: 利用線上樣品濃縮技術偵測對苯二甲酸產品中之不純物 與 毛細管電泳結合質譜儀分析盤尼西林
Development and Application of Capillary Electrophoresis: Determination of Impurity in Terephthalic acid by On-line Concentration and Capillary Electrophoresis Coupled with Mass Spectrometer for Penicillin Analyses
摘要
本論文第一部分為線上樣品濃縮(On-line sample concentration)技 術結合微乳化電層析(MEEKC)技術進行苯甲酸系列化合物的檢測。探 討有無前端溶液、樣品基質等因素對苯甲酸系列化合物的放大效率之 影響。結果顯示，前端酸段溶液注射99 秒及前端水段注射35 秒為最 佳化條件，利用此條件成功的結合微乳化電層析並且有效降低測試分 析物的偵測極限。最後，此技術可成功應用在多種商業上可購得的苯 甲酸系列產品中微量苯甲酸系列的不純物。 毛細管電泳相較於液相層析有較高的分離效率以及低溶劑的使 用量，而質譜儀在近幾年已成為分子結構鑑定最關鍵的工具。論文第 二部份為建立水相，非水相及環糊精修飾的CE方法進行八種盤尼西 林的分析，並測試其與電噴灑質譜儀結合的可行性。實驗中先探討不 同背景電解液組成對測試分析物之分離效率的影響，並最佳化其CE 分離。最後改變背景電解液pH值、離子濃度及有機溶劑比例等因素。 結果顯示，水相背景電解液之最佳分離條件為pH8，60mM 醋酸 銨加10%甲醇做為有機修飾劑；非水相則是以pH6，60mM醋酸銨在 乙腈/甲醇 70/30的比例中為最佳分離條件。但部份分析物仍無法達 到基線分離的效果，故添加不同種類的環糊精測試，發現添加5mM sulfated-CD可得到較好的分離效果。最後測試三種CE模式的最佳條件 串聯電噴灑質譜儀的可行性。
並列摘要
In the first part of the thesis, an on-line sample concentration techniques combined with MEEKC(Microemulsion electrokinetic chromatography) separation to analyze aromatic acid compounds. The effects of several factors such as acid plug solution and sample matrix on the stacking ability of these tested compounds were discussed. The result showed that, acid plug was of 99s and water plug of 35s in the proposed method improved the detection limit of aromatic acid compounds. Finally, this technique has been applied to detect trace-level impurities in commercial aromatic acid compounds. In recent ten years, LC-mass spectrometry (MS) has become a key tool for the characterization of penicillins. In the second part of the thesis, a capillary electrophoresis based on aqueous, nonaqueous or cyclodextrin-modified methods were used to separate eight penicillins, and the possibility of these methods coupled with electrospray ionization/mass spectrometer was discussed. First, the effect of ion type of background electrolyte, types of cyclodextrin and cyclodextrin’s concentration on the separation efficiency of penicillins was discussed. Then, the buffer pH, ionic strength, and portion of organic solvent were examined to optimize the CE separation. The result showed that, the optimal separation conditions was 60mM ammonium acetate solution of pH8 with 10% MeOH for CZE; 60mM ammonium acetate with ACN/MeOH 70/30 at pH6 for NACE; and 5mM sulfated-α-CD for CD-modified CE, respectively. The feasibility of the three optimal CE methods coupled with mass spectrometer was evaluated and compared for the determination of eight penicillins.