A simple and rapid method was presented for the determination of acetylcholine and choline, using the high-performance liquid chromatographic system with enzyme electrodes as the electrochemical detector. The enzyme electrode was made up of cross-linking acetylcholinesterase (AChE) and choline oxidase (ChO) with bovine serum albumin (BSA) or AChE, ChO and peroxidase (POD) with BSA using glutaraldehyde on a platinum electrode silanized with (3-aminopropyl)triethoxysilane. The AChE-ChO and AChE-ChO-POD electrodes were based on the amperometric detection of hydrogen peroxide and hexacyanoferrate(III)generated by the enzymic reactions, respectively. The method was high-sensitive and specific for the determination of acetylcholine and choline. The detection limit was 1.5pmol for choline and 2.9 pmol for acetylcholine. The enzyme electrodes could be used continuously for at least 20 d.

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