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

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 degrees C, respectively. The linear part was in the region 1.0x10(-7)-6.0x10(-7)M (R-2=0.990). Storage stability and operation stability of the enzyme electrode were also studied.