Analysis of serotonin in brain microdialysates using capillary electrophoresis and native laser-induced fluorescence detection.
Serotonin or 5-hydroxytryptamine (5-HT) is a major neurotransmitter in the central nervous system. In this work, a method for analyzing 5-HT in brain microdialysis samples using a commercially available capillary electrophoresis (CE) system has been developed. A pH-mediated in-capillary preconcentration of samples was performed, and after separation by capillary zone electrophoresis, native fluorescence of 5-HT was detected by a 266 nm solid-state laser. The separation conditions for the analysis of 5-HT in standard solutions and microdialysates have been optimized, and this method has been validated on both pharmacological and analytical bases. Separation of 5-HT was performed using a 80 mmol/L citrate buffer, pH 2.5, containing 20 mmol/L hydroxypropyl-beta-cyclodextrin (HP-beta-CD) and +30 kV voltage. The detection limit was 2.5 x 10(-10) mol/L. This method allows the in vivo brain monitoring of 5-HT using a simple, accurate CE measurement in underivatized microdialysis samples.
Benturquia
Nadia
N
Laboratoire de Neuropharmacologie et Neurochimie, INSERM U512, Institut Fédératif des Neurosciences de Lyon (IFR 19), Université Claude Bernard, F-69373 Lyon Cedex 08, France.
Couderc
François
F
Sauvinet
Valérie
V
Orset
Cyrille
C
Parrot
Sandrine
S
Bayle
Christophe
C
Renaud
Bernard
B
Denoroy
Luc
L
D000818:Animals; D001923:Brain Chemistry / Q000187:drug effects; D015283:Citalopram / Q000494:pharmacology; D019075:Electrophoresis, Capillary / Q000379:methods*; D010134:Fenclonine / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007834:Lasers; D008297:Male; D017551:Microdialysis / Q000379:methods; D051381:Rats; D017207:Rats, Sprague-Dawley; D012680:Sensitivity and Specificity; D012701:Serotonin / Q000032:analysis* / Q000302:isolation & purification*; D013050:Spectrometry, Fluorescence / Q000379:methods
D015283:Citalopram; D012701:Serotonin; D010134:Fenclonine