The nano composited film of indigotetrasulfonate (ITS) electrodeposited onto poly-l-lysine (PLL)–glutaraldehyde (GA) (ITS/PLL–GA) was modified on glassy carbon electrode (GCE) by multiple scan cyclic voltammetry. Composited of the proposed film was characterized by atomic force microscopy (AFM), scanning electron microscopy (SEM), electrochemical quartz crystal microbalance (EQCM), electrochemical impedance spectroscopy (EIS), and UV–vis spectrum for the absorption at λ_max at 566 nm. For the electrocatalytic reduction of dissolved oxygen, ITS/PLL–GA film modified electrodes was determined in 0.1 M acetate buffer solution (pH 5.6) by cyclic voltammetry and rotating disk electrode voltammetry. This dissolved oxygen electrochemical sensor exhibited a linear response range (from 0 to 178.4 μM, R² = 0.9949), lowest detection limit (2.2 μM), lowest overpotential at −0.09 V, high sensitivity (906 μA mM⁻¹) and relative standard deviation (RSD) for determining dissolved oxygen (n = 3) was 4.2%. In addition, the ITS/PLL–GA/GCE was advantageous in terms of its simple preparation, specificity, stability and the ability of regeneration.

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