Simultaneous determination of L-glutamate, acetylcholine and dopamine in rat brain by a flow-injection biosensor system with microdialysis sampling

Abstract

A flow-injection biosensor system with an on-line microdialysis sampling system is proposed for the simultaneous detection of neurotransmitters (L-glutamate, acetylcholine and dopamine) released from rat brain cells. The dialysate collected in the sample loop from the microdialysis probe was automatically injected into the flow-injection line with a triple electrode arranged perpendicular to the flow direction. The triple electrode was constructed by hybridizing a poly(1,2-diaminobenzene) film to two enzyme sensing-parts which respond to L-glutamate and acetylcholine, and by coating a Nafion film on a remaining sensing part which responds to dopamine, respectively, without any cross-reactivity. The three sensing parts of the triple electrode responded linearly to the concentrations of L-glutamate and acetylcholine in the range of 0.002-5 mM and to that of dopamine in the range of 0.002-20 mM, respectively, without any interference from oxidizable species present in the dialysate. The proposed flow-injection analytical method could be applied to an in vivo assay of these neurotransmitters released from rat-brain cells by the continuous KCl stimulation.