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A novel H202 sensor was constructed by employing methylene blue (MB) incorporated in a-zirconium phosphate as an electron transfer mediator. Methylene blue was intercalated into a a-zirconium phosphate modified horseradish peroxidase (HRP) immobilization matrix cross-linked on a glassy carbon electrode surface via bovine serum albumin (BSA)-glutaraldehyde. Such a coimmobilization matrix of the mediator and enzyme was formed from a-zirconium phosphate-MB inclusion colloid in which horseradish peroxidase was dissolved. Intercalation of methylene blue in layered a-zirconium phosphate was investigated by X-ray power diffraction (XRD), IR spectra and electrochemical measurements. Methylene blue immobilized in this way underwent a quasi-reversible electrochemical redox reaction. Cyclic voltammetry and amperometric measurements demonstrated that methylene blue coimmobilized with HRP displayed good stability and efficiently shuttled electrons between HRP and the electrode. The sensorresponded rapidly and linearly to HzOz over the range from 2.0 x mo1L Keywords: Amperometric enzyme biosensor, a-Zirconium phosphate, Methylene blue, Hydrogen peroxide to 5 . 0~ mol/L, with a detection limit of 3.0 x

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