Analysis of amino acid neurotransmitters by capillary electrophoresis and laser-induced fluorescence using a new fluorescein-derived label
Analysis of amino acid neurotransmitters by capillary electrophoresis and laser-induced fluorescence using a new fluorescein-derived label
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DOI：
10.1007/s00604-006-0723-6
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发表时间：
2007-05-01
影响因子：
5.7
通讯作者：
Hong, Wang
中科院分区：
文献类型：
--
作者：
Cao, Liwei;Zhang, Huashan;Hong, Wang
建立了一种毛细管电泳激光诱导荧光检测方法（CE-LIF），用于分离用本实验室合成的新型荧光试剂6-氧基-（N-琥珀酰亚胺基乙酸酯）9-（2'-甲氧基羰基）荧光素（SAMF）标记的8种神经递质。衍生化在硼酸缓冲液（pH 7.75）中于37℃进行15分钟。 SAMF 的 pH 依赖性荧光 (pH 4 - 9) 允许背景缓冲液在较宽的 pH 范围内使用。事实证明，与碱性介质相比，酸性电泳缓冲液在分辨率和峰形方面提供了更好的分辨率。使用 Cu2+ 作为添加剂，使用由 40 mM 乙酸钠和 2 mM Cu2+ (pH 6.0) 组成的电泳缓冲液对分子进行基线分离。检测限范围为 1 至 2 x 10(-10) M。该方法已针对淋巴细胞样品的表征进行了验证。获得的结果说明了将 SAMF 衍生化与 CE-LIF 相结合来测定神经递质的优势。
A capillary electrophoresis laser-induced fluorescence detection method (CE-LIF) was developed for the separation of eight neurotransmitters tagged on their amino function with 6-oxy-(N-succinimidyl acetate)9-( 2'-methoxycarbonyl) fluorescein(SAMF), anew fluorescent reagent synthesized in our lab. Derivatization was performed in boric acid buffer ( pH 7.75) at 37 degrees C over 15 min. The pH-independent fluorescence of SAMF ( pH 4 - 9) permits background buffers over a wide range of pH. It was demonstrated that an acidic running buffer offers a better resolution compared to basic medium in terms of resolution and peak shapes. Employing Cu2+ as the additive, the molecules were baseline-separated using a running buffer consisting of 40 mM sodium acetate and 2 mM Cu2+ ( pH 6.0). The detection limits ranged from 1 to 2 x 10(-10) M. The method has been validated for the characterization of lymphocyte samples. The results obtained illustrate the advantages of combining SAMF derivatization with CE-LIF for determining neurotransmitters.