Magnetized nanoporous silica based microparticles (MMPs) were used for horseradish peroxidase (HRP) immobilization and applied for amperometric peroxidase-based biosensor development. A magnetized carbon paste electrode permitted the MMPs attraction. The biosensor was applied to the investigation of the enzymatic oxidation of acetaminophen (paracetamol). The biosensor operated at low applied potential and the signal corresponded to the electroreduction of N-acetylbenzoquinoneimine (NAPQI) generated by the enzyme HRP in the presence of hydrogen peroxide. The biosensor allowed performing the quantitation of acetaminophen in the micromolar concentration range and the comparative study of thiols which inhibited the biosensor response. Distinct inhibition results were observed for HRP entrapped in the silica microparticles compared to the soluble HRP.
The development of a biosensor based on the immobilization of horseradish peroxidase (HRP) within a zirconium alkoxidepolyetilenimine film onto screen-printed electrodes (SPE) for acetaminophen detection and acetaminophen quantification in pharmaceutical products is described. The biosensor operation mode is based on monitoring the amperometric signal produced by the electrochemical reduction of the enzymatically generated electroactive oxidized species of acetaminophen in the presence of hydrogen peroxide. The enzyme immobilization is performed by retention in a polyethylenimine-zirconium alcoxide porous gel film, a technique that offers good entrapping and a protective environment for the biocomponent due to the hydration properties of the immobilization layer. SPEs have the advantage of being easily mass-produced at low costs with superior characteristics in comparison with classical electrode materials. In this configuration, zirconium alkoxide demonstrates its electrocatalytic activity. The biosensor allows the quantification of acetaminophen with a limit of detection of 6.21×10 -8 M and a linear range between 4.35×10 -7 M and 4.98×10 -6 M. Finally, the biosensor is applied to the quantitative analysis of acetaminophen in Perdolan® tablets.
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