Monitoring dopamine in vivo by microdialysis sampling and on-line CE-laser-induced fluorescence.
Microdialysis sampling was coupled on-line to micellar electrokinetic chromatography (MEKC) to monitor extracellular dopamine concentration in the brains of rats. Microdialysis probes were perfused at 0.3 microL/min and the dialysate mixed on-line with 6 mM naphthalene-2,3-dicarboxaldehye and 10 mM potassium cyanide pumped at 0.12 microL/min each into a reaction capillary. The reaction mixture was delivered into a flow-gated interface and separated at 90-s intervals. The MEKC separation buffer consisted of 30 mM phosphate, 6.5 mM SDS, and 2 mM HP-beta-CD at pH 7.4, and the electric field was 850 V/cm applied across a 14-cm separation distance. Analytes were detected by laser-induced fluorescence excited using the 413-nm line of a 14-mW diode-pumped laser. The detection limit for dopamine was 2 nM when sampling by dialysis. The basal dopamine concentration in dialysates collected from the striatum of anesthetized rats was 18 +/- 3 nM (n = 12). The identity of the putative dopamine peak was confirmed by showing that dopamine uptake inhibitors increased the peak and dopamine synthesis inhibitors eliminated the peak. The utility of this method for behavioral studies was demonstrated by correlating dopamine concentrations in vivo and with psychomotor behavior in freely moving rats following the intravenous administration of cocaine. Over 60 additional peaks were detected in the electropherograms, suggesting the potential for monitoring many other substances in vivo by this method.
Shou
Minshan
M
Department of Chemistry, Neuroscience Program, Department of Psychology, and Department of Pharmacology, 930 North University Avenue, University of Michigan, Ann Arbor, Michigan 48109, USA.
Ferrario
Carrie R
CR
Schultz
Kristin N
KN
Robinson
Terry E
TE
Kennedy
Robert T
RT
D000818:Animals; D001921:Brain / Q000378:metabolism; D020374:Chromatography, Micellar Electrokinetic Capillary / Q000379:methods*; D004298:Dopamine / Q000378:metabolism*; D005453:Fluorescence; D007834:Lasers; D008297:Male; D017551:Microdialysis; D051381:Rats; D017207:Rats, Sprague-Dawley
D004298:Dopamine