Development of an acetylcholinesterase–choline oxidase based biosensor for acetylcholine determination

Abstract

In this article, a novel amperometric acetylcholine (ACh) biosensor with immobilization of acetylcholinesterase–choline oxidase (AChE–ChO) on electrochemically polymerized polyaniline–polyvinylsulphonate (PANI–PVS) film has been successfully developed for the first time. AChE and ChO enzymes were immobilized on PANI–PVS film by cross-linking with using glutaraldehyde (GA) and bovine serum albumin (BSA). Determination of ACh was carried out by oxidation of enzymatically produced H2O2 at +0.4 V versus Ag/AgCl. Optimum pH and temperature were found to be 8.0 and 60 °C, respectively. The linear part was in the region 1.0 × 10−7–6.0 × 10−7 M (R2 = 0.990). Storage stability and operation stability of the enzyme electrode were also studied.