Chiral Measurement of Aspartate and Glutamate in Single Neurons by Large-Volume Sample Stacking Capillary Electrophoresis

Amit V. Patel, Takayuki Kawai, Liping Wang, Stanislav S. Rubakhin, Jonathan V. Sweedler

Research output: Contribution to journalArticle

Abstract

d-Amino acids (d-AAs) are endogenous molecules found throughout the metazoan, the functions of which remain poorly understood. Measurements of low abundance and heterogeneously distributed d-AAs in complex biological samples, such as cells and multicellular structures of the central nervous system (CNS), require the implementation of sensitive and selective analytical approaches. In order to measure the d- and l-forms of aspartate and glutamate, we developed and applied a stacking chiral capillary electrophoresis (CE) with laser-induced fluorescence detection method. The achieved online analyte preconcentration led to a 480-fold enhancement of detection sensitivity relative to capillary zone electrophoresis, without impacting separation resolution or analysis time. Additionally, the effects of inorganic ions on sample preconcentration and CE separation were evaluated. The approach enabled the relative quantification of d-aspartate and d-glutamate in individual neurons mechanically isolated from the CNS of the sea slug Aplysia californica, a well characterized neurobiological model. Levels of these structurally similar d-AAs were significantly different in subpopulations of cells collected from the investigated neuronal clusters.

Original languageEnglish (US)
Pages (from-to)12375-12382
Number of pages8
JournalAnalytical chemistry
Volume89
Issue number22
DOIs
StatePublished - Nov 21 2017

ASJC Scopus subject areas

  • Analytical Chemistry

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