Biorecognition of hydrogen peroxide using a novel electrochemical platform designed with Glossoscolex paulistus giant...

The giant extracellular hemoglobin of the annelid Glossoscolex paulistus (HbGp; 3.6 MDa) is a valuable and underexplored supramolecular hemoprotein system for the biorecognition of reactive oxygen species. In this work, an efficient and simple electrochemical platform was designed for analyzing H₂O₂, using HbGp covalently immobilized on Nafion®-modified glassy carbon electrode, named as HbGp/Nafion/GCE. Voltammetric and spectroscopic studies revealed the importance of prior modification of the electrodic support with the conducting polymer to obtain satisfactory hemoglobin electroactivity, as well as a biocompatible microenvironment for its immobilization. In terms of biological activity, it was observed a greater reactivity of the biomolecule in acidic medium, enabling the detection of the analyte by a quasi-reversible mechanism, whose kinetics was limited by analyte diffusion. In the presence of H₂O₂, the native structure of hemoglobin (oxy-HbGp (Fe²⁺)) oxidizes to ferryl-HbGp (Fe⁴⁺) and this redox reaction can be monitored on HbGp/Nafion/GCE with a detection limit of 8.5 × 10^‒7  mol L^-1. In addition to high sensitivity, the electrochemical biosensor also provided reproducible, consistent, and accurate measurements. The electroanalytical method showed an appropriate performance to quantify different levels of H₂O₂ in milk samples, proving the potential of HbGp/Nafion/GCE for this purpose.