We present here new amperometric electrodes for the selective and quantitative detection of acetylcholine. The detection was achieved based on the electrodeposition of a carbon electrode with poly(3,4-ethylenedioxythiophene) (PEDOT) followed by the drop-casting of an ionophore-doped poly(vinyl) chloride (PVC) membrane. This work paves the way for future applied research to study neurological disorders.