DETERMINATION OF ACETYLCHOLINE BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY, ENZYME-LOADED POSTCOLUMN,AND ELECTROCHEMICAL DETECTOR

A rapid and simple method using a combination of high performance liquid chromatography (HPLC), an immobilized enzyme column, with electrochemical detection (ECD) is described for the assay of acetylcholine (ACh) and choline (Ch) in mouse brain. Perchloric acid extracts of small brain tissue were injected onto the HPLC system with no preclean-up procedure. Detection limits for both compounds were 1 pmol. The present method has been successfully applied to the measurement of ACh in discrete brain areas of the mouse.