Optimization of TEMPO-Mediated Oxidation of Chitosan to Enhance Its Antibacterial and Antioxidant Activities

Abstract

This study systematically investigated the oxidation of chitosan using the TEMPO/NaClO/NaBr catalytic system under varying experimental conditions, namely temperature, reaction time, and pH, in order to optimize the oxidation process. Response surface methodology (RSM) was employed to determine the optimal parameters for maximizing the efficiency of the reaction. The structural modifications to the chitosan following oxidation were confirmed using Fourier-transform infrared spectroscopy (FTIR), alongside additional analytical techniques, which validated the successful introduction of carbonyl and carboxyl functional groups. Solvent-cast films were prepared from both native and oxidized chitosan in order to evaluate their functional performance. The antibacterial activity of these films was assessed against Gram-negative (Salmonella) and Gram-positive (Streptococcus faecalis) bacterial strains. The oxidized chitosan films exhibited significantly enhanced antibacterial effects, particularly at shorter incubation periods. In addition, antioxidant activity was evaluated using DPPH radical scavenging and ferrous ion chelation assays, which both revealed a marked improvement in radical scavenging ability and metal ion binding capacity in oxidized chitosan. These findings confirm that TEMPO-mediated oxidation effectively enhances the physicochemical and bioactive properties of chitosan, highlighting its potential for biomedical and environmental applications.