Determination of kynurenic acid by capillary electrophoresis with laser-induced fluorescence detection.
Kynurenic acid (KA) is an excitatory amino acid receptor antagonist that is believed to play an important role in a host of diseases of the neuropsychiatric and central nervous system. A method for the determination of KA in microdialysate samples using capillary electrophoresis (CE) separation with laser-induced fluorescence (LIF) detection is described. CE is advantageous for the analysis of microdialysis samples due to its short analysis times and small sample volume requirements. Three complexation approaches were evaluated in an attempt to achieve the best limit of detection. The best approach was found to be pre-column complexation with inclusion of Zn(II) in the background electrolyte. After optimization of the zinc acetate concentration and pH, a limit of detection of 1 nM KA was achieved. However, when KA was present in the dialysate, the limit of detection increased 50-fold. Even though the endogenous levels of KA in rat brain are below this limit of detection, this methodology could be used to monitor the increase of KA levels in rat brain following dosing with its precursors, tryptophan and kynurenine.
Hansen
D K
DK
Department of Chemistry, University of Kansas, Lawrence 66047, USA.
Lunte
S M
SM
D000818:Animals; D019075:Electrophoresis, Capillary / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D007274:Injections, Intraperitoneal; D007736:Kynurenic Acid / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D007737:Kynurenine / Q000008:administration & dosage / Q000378:metabolism / Q000493:pharmacokinetics*; D009994:Osmolar Concentration; D051381:Rats; D013050:Spectrometry, Fluorescence; D013997:Time Factors; D014364:Tryptophan / Q000008:administration & dosage / Q000378:metabolism / Q000493:pharmacokinetics*; D015032:Zinc / Q000737:chemistry*
D007737:Kynurenine; D014364:Tryptophan; D007736:Kynurenic Acid; D015032:Zinc