In this work, carbon electrodes modified with aminophenols were developed for the production of pesticides biosensors based on acetylcholinesterase. The polymers were potentiodynamically deposited on a graphite electrode surface by the oxidation of monomers, 2-aminophenol, 3-aminophenol and 4-aminophenol. The electrochemical behaviour and surface analysis of the electrodes modified by polyaminophenols non-immobilized and immobilized on acetylcholinesterase were studied by cyclic voltammetry, electrochemical impedance spectroscopy and atomic force microscopy. Roughness values obtained for graphite electrodes modified with poly(4-aminophenol) and poly(4-aminophenol)/acetylcholinesterase were 174 and 86 nm, respectively. The acetylcholinesterase enzyme was immobilized on a graphite and a graphite modified with poly(4-aminophenol), and these electrodes were coupled in the flow system. Potentiometric response due to hydrogen ions generated by an enzymatic system in the presence of acetylcholine chloride substrate was evaluated. The results showed that the graphite/poly(4-aminhophenol) sensor presents high sensitivity to hydrogen ions when compared with other graphite/polyaminophenols sensors. The biosensor coupled in a continuous flow system was employed for the detection of dichlorvos. The detection and quantification limits were 0.8 and 2.4 μmol L −1 dichlorvos, respectively. This sensor reveals an efficient and promising material for biomolecules immobilization.