A microchip electrophoresis-mass spectrometric platform for fast separation and identification of enantiomers employing the partial filling technique.
A microchip electrophoresis-mass spectrometric platform for fast separation and identification of enantiomers employing the partial filling technique.
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DOI：
10.1016/j.chroma.2013.10.020
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发表时间：
2013-11-29
期刊：
影响因子：
--
通讯作者：
Liu YM
中科院分区：
文献类型：
--
作者：
Li X;Xiao D;Ou XM;McCullm C;Liu YM
建立了一种快速手性分析的微芯片电泳质谱(MCE-MS)方法。所提出的MCE-MS平台部署了玻璃/PDMS混合微芯片和易于制造的单片式纳米电喷雾发射器。采用部分填充技术实现了对映体MCE的分离。一种新颖的芯片设计，其中臂通道连接到MCE分离通道的中间，用于传递手性选择器，并被测试和证明是有效的。3，4-二羟基苯丙氨酸(DOPA)、谷氨酸(Glu)和丝氨酸(Ser)在S范围内实现了对映体分离，分离度分别为2.4、1.1和1.0。该方法灵敏度高，L-多巴和D-多巴的检出限分别为43 nM和47 nM。该分析平台集细胞培养、进样、手性分离和MS检测于一体，非常适合于活细胞立体化学偏好的研究。以DOPA在人SH-SY5Y神经细胞中的代谢为模型系统进行了研究。将SH-SY5Y细胞与外消旋多巴进行芯片孵育，每隔一段时间注入孵育液并进行在线检测。结果发现，随着孵育时间的延长，L-多巴的浓度逐渐降低，而共存的D-多巴的浓度保持不变。结果表明，SH-SY5Y细胞能有效代谢L-多巴，而D-多巴保持不变。
A microchip electrophoresis-mass spectrometric (MCE-MS) method was developed for fast chiral analysis. The proposed MCE-MS platform deployed a glass /PDMS hybrid microchip with an easy-to-fabricate monolithic nanoelectrospray emitter. Enantiomeric MCE separation was achieved by means of the partial filling technique. A novel chip design with an arm channel connecting to the middle of the MCE separation channel for delivering the chiral selector was tested and proven valid. Enantiomeric separation of 3.4-dihydroxyphenylalanine (DOPA), glutamic acid (Glu), and serine (Ser), the selected test compounds, were achieved within 130 s with resolution values (Rs) of 2.4, 1.1, and 1.0, respectively. The proposed chiral MCE-MS assay was sensitive and had detection limits of 43 nM for L-DOPA and 47 nM for D-DOPA. The analytical platform was well suited for studies of stereochemical preference in living cells because it integrated cell culture, sample injection, chiral separation, and MS detection into a single platform. Metabolism of DOPA in human SH-SY5Y neuronal cells was studied as a model system. On-chip incubation of SH-SY5Y cells with racemic DOPA was carried out, and the incubation solution was injected and in-line assayed at time intervals. It was found that L-DOPA concentration decreased gradually as incubation time increased while the concentration of coexisting D-DOPA remained constant. The results firmly indicated that SH-SY5Y cells metabolized L-DOPA effectively while left D-DOPA intact.
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