Simple method for the simultaneous determination of acetylcholine, choline, noradrenaline, dopamine and serotonin in brain tissue by high-performance liquid chromatography with electrochemical detection.
A simple method for the simultaneous determination of acetylcholine, choline, noradrenaline, dopamine and serotonin in brain tissue was developed by using high-performance liquid chromatography with electrochemical detection. These compounds are analysed in a single chromatographic run within 30 min with a simple sample clean-up procedure. The detection system consists of two electrochemical detector cells aligned in series: a glassy-carbon electrode for catecholamines and serotonin, and a platinum electrode for acetylcholine and choline. For the detection of the latter compounds, they were converted enzymatically into hydrogen peroxide through a column reactor with immobilized acetylcholinesterase and choline oxidase. A column of boronic acid gel was placed just ahead of the immobilized enzyme column to remove catecholamines, which caused interfering responses on the platinum electrode. Two equivalent analytical columns and a column switching were employed to speed up the serotonin assay. Simultaneous determination of these major neurotransmitters in rat brain regions was successfully carried out with the system described.
D000109:Acetylcholine / Q000032:analysis; D000818:Animals; D001679:Biogenic Amines / Q000032:analysis*; D001923:Brain Chemistry; D002794:Choline / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D004298:Dopamine / Q000032:analysis; D004563:Electrochemistry; D008297:Male; D009638:Norepinephrine / Q000032:analysis; D051381:Rats; D012701:Serotonin / Q000032:analysis
D001679:Biogenic Amines; D012701:Serotonin; D002794:Choline; D000109:Acetylcholine; D004298:Dopamine; D009638:Norepinephrine