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showed that different sizes of iron oxide nanoparticles (30, 50, and 300 nm) could oxidize the colorless TMB into a blue colored product in the presence of hydrogen peroxide at acidic pH (Gao et al., Further, the Km value of iron oxide nanoparticles with TMB was about four times lesser than HRP (0.098 and 0.43 mM, respectively), suggesting that nanoparticles have a higher affinity for the substrate (TMB) than HRP, therefore, at the same molar concentrations, nanoparticles showed 40 times higher activity than HRP