In vivo monitoring of amine neurotransmitters using microdialysis with on-line capillary electrophoresis.
Microdialysis sampling was coupled via a flow-gated interface on-line to capillary electrophoresis with laser-induced fluorescence (LIF) detection for in vivo monitoring of neuroactive amino acids and amines. In the instrument, analytes are derivatized precolumn with o-phthaldehyde and beta-mercaptoethanol to form fluorescent isoindole products. The instrument was improved over previous designs by incorporating a sheath-flow cuvette for reduced background in LIF detection which improved sensitivity by 15-fold. The methodology was improved by utilizing a voltage ramped injection which allowed generation of 500000 theoretical plates with 20 s separations. Resolution of the isoindole derivatives was further improved by addition of hydroxypropyl-modified beta-cyclodextrin to the electrophoresis buffer. The new instrumentation and methods allow resolution and detection of glutamate, gamma-aminobutyric acid, glycine, aspartate, serine, taurine, glutamine and dopamine (if levels are elevated) collected from in vivo sampling probes every 20 s. The technique is suited to continuous monitoring for dynamic measurements of these compounds in vivo.
Bowser
M T
MT
University of Florida, Department of Chemistry, Gainesville, USA.
Kennedy
R T
RT
D000818:Animals; D001923:Brain Chemistry; D004298:Dopamine / Q000032:analysis; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods; D004867:Equipment Design; D008297:Male; D017551:Microdialysis / Q000295:instrumentation / Q000379:methods; D008991:Monitoring, Physiologic / Q000295:instrumentation / Q000379:methods; D018377:Neurotransmitter Agents / Q000032:analysis*; D009862:Online Systems; D051381:Rats; D017207:Rats, Sprague-Dawley; D012680:Sensitivity and Specificity; D013654:Taurine / Q000032:analysis; D013997:Time Factors; D005680:gamma-Aminobutyric Acid / Q000032:analysis
D018377:Neurotransmitter Agents; D013654:Taurine; D005680:gamma-Aminobutyric Acid; D004298:Dopamine