A high-throughput on-line microdialyis-capillary assay for D-serine

Kylie B. O'Brien, Manuel Esguerra, Christina T. Klug, Robert F. Miller, Michael T. Bowser

Research output: Contribution to journalArticle

56 Scopus citations

Abstract

A high-throughput method is described for the analysis of D-serine and other neurotransmitters in tissue homogenates. Analysis is performed by microdialysis-capillary electrophoresis (CE) with laser-induced fluorescence (LIF) detection in a sheath flow detection cell. Sample pretreatment is not required as microdialysis sampling excludes proteins and cell fragments. Primary amines are derivatized on-line with o-phthaldialdehyde (OPA) in the presence of β-mercaptoethanol followed by on-line CE-LIF analysis. Under the separation conditions described here, D-serine is resolved from L-serine and other primary amines commonly found in biological samples. Each separation requires less than 22 s. Eliminating the need for sample pretreatment and performing the high-speed CE analysis on-line significantly reduces the time required for D-serine analysis when compared with traditional methods. This method has been used to quantify D-serine levels in larval tiger salamander retinal homogenates, as well as dopamine, γ-amino-n-butyric acid (GABA), glutamate and L-aspartate. D-serine release from an intact retina was also detected.

Original languageEnglish (US)
Pages (from-to)1227-1235
Number of pages9
JournalELECTROPHORESIS
Volume24
Issue number7-8
DOIs
StatePublished - Apr 2003

Keywords

  • Amino acid detection
  • Enantiomer separation
  • Neurotransmitter
  • Retina

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