On the Use of a TiO2 Sol-Gel for Horseradish Peroxidase Enzyme Immobilization for Development of an Amperometric Biosensor

Abstract

The sol-gel technique has drawn considerable attention in the area of biomolecules immobilization by virtue of its intrinsic properties including chemical inertness, thermal stability, and good biocompatibility. The present work reports the development of a bio-sensing methodology which involves the immobilization of horseradish peroxidase enzyme (HRP) on pencil graphite electrode (PGE) using TiO2 sol-gel and its application for 5-aminosalicylic acid (5-ASA) determination in pharmaceutical formulations. TiO2 sol-gel film was deposited on the PGE surface by dip coating, and the HRP enzyme was immobilized on the film by adsorption and cross-linked binding using glutaraldehyde. The analytical performance of the methodology was investigated through amperometric measurements at –0.08 V in the presence of 5-ASA (electron mediator) and hydrogen peroxide (substrate). The biosensor was successfully applied for 5-ASA determination, where limits of detection and quantification of 3.3 and 10.0 μmol L-1, respectively, were obtained. The application of the methodology for 5-ASA determination in pharmaceutical formulations led to satisfactory recovery rates ranging from 80 to 98%. The biosensor developed in this work is simple, inexpensive and has the potential to be applied for the determination of other pharmaceutical compounds, which also act as electron mediators in the catalytic cycle of HRP.