Biofunctionalisation of Polypyrrole Nanowires Array with Sulfite Oxidase Coupled with the Integration of Platinum Nanoparticles for Ultrasensitive Amperometric Detection of Sulfite.

Sulfite determination in foods and alcoholic beverages is a common requirement by food and drug administration organisations in most countries. In this study, the enzyme, sulfite oxidase (SOx), is used to biofunctionalise a platinum-nanoparticle-modified polypyrrole nanowire array (PPyNWA) for the ultrasensitive amperometric detection of sulfite. A dual-step anodisation method was used to prepare the anodic aluminum oxide membrane used as a template for the initial fabrication of the PPyNWA. PtNPs were subsequently deposited on the PPyNWA by potential cycling in a platinum solution. The resulting PPyNWA-PtNP electrode was then biofuntionalised by adsorption of SOx onto the surface. The confirmation of the adsorption of SOx and the presence of PtNPs in the PPyNWA-PtNPs-SOx biosensor was verified by scanning electron microscopy and electron dispersive X-ray spectroscopy. Cyclic voltammetry and amperometric measurements were used to investigate the properties of the nanobiosensor and to optimise its use for sulfite detection. Ultrasensitive detection of sulfite with the PPyNWA-PtNPs-SOx nanobiosensor was accomplished by use of 0.3 M pyrrole, 10 U mL-1 of SOx, adsorption time of 8 h, a polymerisation period of 900 s, and an applied current density of 0.7 mA cm-2. The response time of the nanobiosensor was 2 s, and its excellent analytical performance was substantiated with a sensitivity of 57.33 μA cm-2 mM-1, a limit of detection of 12.35 nM, and a linear response range from 0.12 to 1200 μM. Application of the nanobiosensor to sulfite determination in beer and wine samples was achieved with a recovery efficiency of 97-103%.