Development of a PDMS-based microchip electrophoresis device for continuous online in vivo monitoring of microdialysis samples.
A PDMS-based microfluidic system for online coupling of microdialysis sampling to microchip electrophoresis with fluorescence detection for in vivo analysis of amino acid neurotransmitters using naphthalene-2,3-dicarboxaldehyde and sodium cyanide as the derivatization reagents is described. Fabricating chips from PDMS rather than glass was found to be simpler and more reproducible, especially for chips with complex designs. The microchip incorporated a 20-cm serpentine channel in which sample plugs were introduced using a "simple" injection scheme; this made fluid handling and injection on-chip easier for the online system compared with gated or valve-based injection. The microchip was evaluated offline for the analysis of amino acid standards and rat brain microdialysis samples. Next, precolumn derivatization was incorporated into the chip and in vivo online microdialysis-microchip electrophoresis studies were performed. The system was employed for the continuous monitoring of amino acid neurotransmitters in the extracellular fluid of the brain of an anesthetized rat. Fluorescein was dosed intravenously and monitored simultaneously online as a marker of in vivo blood-brain barrier permeability. The microdialysis-microchip electrophoresis system described here will be employed in the future for simultaneous monitoring of changes in blood-brain barrier permeability and levels of amino acid neurotransmitters in the rat stroke model.
Nandi
Pradyot
P
Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, KS, USA.
Desai
Dhara P
DP
Lunte
Susan M
SM
D000596:Amino Acids / Q000032:analysis*; D000818:Animals; D001923:Brain Chemistry; D004129:Dimethylpolysiloxanes / Q000737:chemistry*; D046209:Electrophoresis, Microchip / Q000295:instrumentation* / Q000379:methods*; D019793:Fluorescein / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D017551:Microdialysis / Q000379:methods*; D009281:Naphthalenes / Q000737:chemistry; D018377:Neurotransmitter Agents / Q000032:analysis*; D009757:Nylons / Q000737:chemistry*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012966:Sodium Cyanide / Q000737:chemistry; D013050:Spectrometry, Fluorescence / Q000379:methods
D000596:Amino Acids; D004129:Dimethylpolysiloxanes; D009281:Naphthalenes; D018377:Neurotransmitter Agents; D009757:Nylons; C085707:poly(dimethylsiloxane)-polyamide copolymer; C027717:2,3-naphthalenedicarboxaldehyde; D012966:Sodium Cyanide; D019793:Fluorescein