单细胞中D-天冬氨酸和D-谷氨酸的毛细管电泳分析

Analysis of D-Aspartic Acid and D-Glutamic Acid in Single Cells by Capillary Electrophoresis

建立了一种毛细管电泳-激光诱导荧光检测单个神经细胞中D-天冬氨酸和D-谷氨酸的方法.以萘二甲醛(NDA)为衍生试剂,β-环糊精(β-CD)和脱氧胆酸钠(SDC)为拆分体系,对16对氨基酸对映体的NDA衍生物进行了手性拆分研究.考察了蛋白氨基酸和一些胺对检测D-天冬氨酸和D-谷氨酸的影响,并确立了最佳分离条件,从而建立了单细胞中D-天冬氨酸和D-谷氨酸的检测方法,并对海兔的单个神经细胞中D-天冬氨酸和D-谷氨酸进行了检测.

An analytical method was established to detect D-aspartic acid（D-Asp） and D-glutamic acid（D- Glu） in single cells. Cellular samples were derivatized with a fluorescence tagging reagent, naphthalene-2,2- dicarboxaldehyde in a mini-volume（3 μL） prior to the separation. Sixteen pair of amino acid enantiomers resolution was achieved by using a dual chiral selector system consisting of β-cyclodextrin（β-CD） and chiral micelles formed by sodium deoxycholate （SDC）. The separation was coupled with laser induced fluorescence detection. Single neurons isolated from the ganglia of Aplysia were analyzed to determine the D/（ D ＋ L） molar ratios of Asp to Glu. D-Asp and D-Glu were found at high levels and its cellular distribution was highly heterogeneous.