High-efficiency application of CTS-Co NPs mimicking peroxidase enzyme on TMB(ox)

Abstract

In this study, analytical studies of Chitosan-Cobalt(II) (CTS-Co(II)) nanoparticles (CTS - Co NPs) by mimicking horseradish peroxidase (HRP) were evaluated. In the applications, it was observed that CTS-Co NPs 3,3' 5,5' tetramethylbenzidine (TMB) oxidized in the presence of hydrogen peroxide (H₂O₂). The required CTS-Co NPs were synthesized at 50 °C in 30 min and characterized using Fourier transform infrared spectroscopy (FTIR), thermal gravimetric analysis (TGA), inductively coupled plasma-optical emission spectroscopy (ICP-OES), and X-ray photon spectroscopy (XPS) was done. CTS-Co NPs were studied to develop a selective TMB biosensor on TMB(ox) substrate. The synthesized CTS-Co NPs formed a catalytic reaction with 30% 0.2 mM H₂O₂ on 0.2 M TMB substrate. After the catalytic reaction, sensitive signals were obtained from the desired biosensor. Electrochemical measurements were taken as low limit of 10 mg and a high limit of 20 mg for the determination of CTS-Co NPs to TMB(ox). In the microplate study; The sensors were applied on 1.5 μg and 3 μg CTS-Co NPs TMB(ox) substrate, respectively. CTS- Co NPs; for TMB(ox) determination, optical density (OD) measurement was taken as a low limit of 1.5 μg and a high limit of 3 μg. Electrochemical applications of particles and microplate reader results were compared with horseradish peroxidase (HRP) enzyme for sensor properties. According to the data obtained, it was observed that it behaved similarly to the CTS-Co NPs peroxidase enzyme. This work presents innovations for nanoparticle extraction and sensor study from chitosan and other naturally sourced polymers.

Keywords: 3,3′ 5,5′ tetramethylbenzidine; Biosensor; Chitosan-cobalt(II); Electrochemistry; Horsearadish peroxidase.