Comparison of quantitative performance of three fluorescence labels in CE/LIF analysis of aspartate and glutamate in brain microdialysate

Zsolt Wagner, Tamás Tábi, Gergely Zachar, András Csillag, Éva Szöko

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Three different fluorescent tags have been compared for the quantitative analysis of aspartate and glutamate in brain microdialysate samples. Separation conditions have been optimized to achieve short analysis time using reversed polarity separation in coated capillary. Method validation has revealed similar quantification limit of 0.1μM of analytes using either of the labels, although LOD values were different: 7.8-9.8nM for 4-fluoro-7-nitro-2,1,3-benzoxadiazole, 3.5nM for fluorescein-5-isothiocyanate and 1.3-1.5nM for carboxyfluorescein succinimidyl ester derivatives. The almost two orders of magnitude difference between LOD and LOQ values is likely due to the unreliable derivatization reaction at low sample concentration. Based on the superior stability, FITC derivatization was used for the analysis of biological samples. The applicability of the method has been demonstrated by analyzing basal and potassium evoked amino acid concentrations in individual brain microdialysate samples.

Original languageEnglish
Pages (from-to)2816-2822
Number of pages7
JournalELECTROPHORESIS
Volume32
Issue number20
DOIs
Publication statusPublished - Oct 1 2011

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Keywords

  • CE-LIF
  • CFSE
  • Excitatory amino acids
  • FITC
  • NBD-F

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry

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