Detection of basal acetylcholine release in the microdialysis of rat frontal cortex by high-performance liquid chromatography using a horseradish peroxidase-osmium redox polymer electrode with pre-enzyme reactor.
To determine the basal acetylcholine level in the dialysate of rat frontal cortex, a horseradish peroxidase-osmium redox polymer-modified glassy carbon electrode (HRP-GCE) was employed instead of the conventional platinum electrode used in high-performance liquid chromatography-electrochemical detection (HPLC-ED). In initial experiments, an oxidizable unknown compound interfered with the detection of basal acetylcholine release on HPLC-HRP-GCE. An immobilized peroxidase-choline oxidase precolumn (pre-reactor) was included in the HPLC system, to eliminate the interference from the unknown compound. This combination could detect less than 10 fmol of standard acetylcholine and basal acetylcholine levels in the dialysate from a conventional concentric design microdialysis probe, without the use of cholinesterase inhibitor, and may facilitate physiological investigation of cholinergic neuronal activity in the central nervous system.
Kato
T
T
Laboratory of Molecular Recognition, Graduate School of Integrated Science, Yokohama City University, Japan.
Liu
J K
JK
Yamamoto
K
K
Osborne
P G
PG
Niwa
O
O
D000109:Acetylcholine / Q000032:analysis* / Q000378:metabolism; D000115:Acetylesterase / Q000378:metabolism; D000429:Alcohol Oxidoreductases / Q000378:metabolism; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004563:Electrochemistry; D004566:Electrodes; D005625:Frontal Lobe / Q000378:metabolism*; D006735:Horseradish Peroxidase / Q000378:metabolism; D016014:Linear Models; D008297:Male; D017551:Microdialysis; D009992:Osmium / Q000737:chemistry; D010084:Oxidation-Reduction; D051381:Rats; D012680:Sensitivity and Specificity
D009992:Osmium; D000429:Alcohol Oxidoreductases; C019888:choline oxidase; D006735:Horseradish Peroxidase; D000115:Acetylesterase; D000109:Acetylcholine