A novel hypoxanthine biosensor fabricated by immobilizing the xanthine oxidase (XOD) onto the phosphonic acid-functionalized silica (SiO2-P) film on the surface of glassy carbon electrode (GCE) was designed and constructed in this work. A biomimetic platform was designed with the phosphonic acid-functionalized silica nanoparticles (SiO2-P NPs) synthesized by the method of reverse microemulsion and electrostatic binding. In such a platform, XOD was selected as model protein to fabricate hypoxanthine biosensor based on SiO2-P NPs. The nanocomposite was characterized with transmission electron microscopy (TEM), energy dispersive X-ray spectrometer (EDS) and electrochemical impedance spectroscopy (EIS). Based on the advantageous functions of SiO2-P NPs, the entrapped XOD could preserve its bioactivity and exhibited an excellent electrochemical behavior with a formal potential of -0.37 V in phosphate buffer solution (PBS, pH=7). Response studies to hypoxanthine were carried out using current-time response curve. The biosensor exhibited a wide linear response ranging from 1.00×10(-6) to 2.61×10(-4) M. The detection limit of 2.33×10(-7) M at a signal-to-noise ratio of 3 was lower than that most reported previously. In addition, the electrode modified with XOD/(SiO2-P NPs) film also had a strong anti-interference ability in the presence of uric acid (UA) and ascorbic acid (AA). The assay results of hypoxanthine in fish samples were in a good agreement with the reference values.
Keywords: Biomimetic; Biosensor; Phosphonic acid-functionalized; Silica; Xanthine oxidase.
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