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題 名 | 以微胞電動層析與反向電滲流同步分析myo-肌醇與D-chiro-肌醇=Simultaneous Determination of myo-Inositol and D-chiro-Inositol by Micellar Electrokinetic Chromatography with Reversed Electroosmotic Flow |
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作 者 | 葉于菁; 曾惠芬; | 書刊名 | 化學 |
卷 期 | 73:1 2015.03[民104.03] |
頁 次 | 頁39-46 |
專 輯 | 生物分析化學 |
分類號 | 341 |
關鍵詞 | myo-肌醇; D-chiro-肌醇; 微胞電動層析; myo-inositol; D-chiro-inositol; Micellar electrokinetic chromatography; |
語 文 | 中文(Chinese) |
中文摘要 | 本文介紹一個簡單的微胞電動層析法 (MEKC),以同步分析兩個肌醇異構物:myo-肌醇與 D-chiro肌醇。電泳分離之前,肌醇需在 25 °C下、與 N-methylisatoic anhydride進行 10分鐘的衍生反應,以利後續分離的螢光檢測。樣品稀釋液中的硼砂 (borax) 可與肌醇衍生物錯合成負離子,再藉由添加溴化十六烷基三甲基銨 (cetyltrimethylammonium bromide, CTAB) 於電泳緩衝液 MES-NaOH中造成的反向電滲流、選擇性電動進樣;樣品緩衝液與電泳緩衝液間的酸鹼度差則促使樣品堆積,尤其是 myo肌醇。本文探討影響分離的參數包括:電泳緩衝液的 pH值、MES與 CTAB的濃度,以及分離電壓。此 MEKC法可於 17分鐘內完成;以甘油為內標準品,myo-肌醇與 D-chiro-肌醇的線性範圍分別為 0.003-0.5 mM與 0.03-1 mM、偵測極限分別是 0.26 μM及 18 μM。與先前的 MEKC法相較,本文介紹的 MEKC法可使 myo-肌醇的偵測靈敏度提升 80倍。 |
英文摘要 | A simple micellar electrokinetic chromatography (MEKC) method was developed for the simultaneous determination of myo-inositol and D-chiro-inositol. Prior to electrophoretic separation, the non-fluorescent inositols were derivatized by N-methylisatoic anhydride at 25 °C for 10 min to allow for fluorescence detection during separation. An MES-NaOH buffer containing cetyltrimethylammonium bromide (CTAB) was used as the background electrolyte (BGE). Borax was added into the sample dilution buffer to facilitate the formation of negatively charged complexes with inositol derivatives. The reversed electroosmotic flow enhanced the electrokinetic injection of negatively charged analytes, and the pH junction between the sample buffer and the BGE resulted in sample stacking, especially for myo-inositol. This good separation with high efficiency by MEKC was achieved in 17 min. Several parameters affecting the separation were studied, including the pH of BGE, the concentrations of MES and CTAB, and the applied voltage. When glycerol was used as an internal standard, the linear ranges of the method for myo-inositol and Dchiro-inositol were 0.003-0.5 and 0.03-1 mM, respectively; the limits of detection of myo-inositol and D-chiro-inositol were 0.26 and 18 μM, respectively. The method was applied to the analysis of human urinary myo-inositol and D-chiro-inositol. Compared to the conventional MEKC method, this proposed method enabled an 80-fold increase in the sensitivity for myo-inositol. |
本系統中英文摘要資訊取自各篇刊載內容。