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The anthraquinone-2,6-disulfonate (AQDS) and poly-l-lysine (PLL) - glutaraldehyde (GA) film (AQDS/PLL-GA) was prepared on a glassy carbon electrode (GCE) by multiple scan cyclic voltammetry. This nanocomposite film was characterized using atomic force microscopy, scanning electron microscopy and electrochemical impedance spectroscopy. The advantages of the film were demonstrated for the determination of S2O82-, IO3- and dissolved oxygen using cyclic voltammetry and rotating disk electrode voltammetry. The electrocatalytic reduction of S2O82-, IO3- and dissolved oxygen at different electrode surfaces, such as bare GCE, PLL-GA/GCE and AQDS/PLL-GA/GCE modified electrodes, was determined in 0.1 M PBS (pH 7.0). The electrochemical sensor exhibited wide linear response range, low detection limit, high sensitivity and good stability.