Abstract: A biosensor based on glucose oxidase (GOx) immobilized onto nanoparticles zirconium oxide with polyethylene nanocomposite for glucose monitoring has been designed. The CTAB/PEG/ZrO2/GOx nanocomposite was deposited onto screen printed carbon paste (SPCE) electrode via spin coating technique. The properties of CTAB/PEG/ZrO2/GOx were study using scanning electron microscopy (SEM). The SPE modified with the CTAB/PEG/ZrO2/GOx showed electrocatalytical response to the oxidation of glucose when ferrocene carboxaldehyde was used as an artificial redox mediator, which was studied by cyclic voltammetry (CV). Several parameters such as working potential, effect of pH and effect of ZrO2/PEG layers that governed the analytical performance of the biosensor, have been studied. The biosensor was applied to detect glucose with a linear range of 0.4 to 2.0 mmol L−1 with good repetability and reproducibility.