Old dogs, new trick: Classical starch˗sodium iodide reagent as the chromogenic substrate of peroxidase-like nanozymes

2, 2′-azinobis-3-ethylbenzthiazoline-6-sulfonate (ABTS), 3, 3′, 5, 5′-tetramethylbenzidine (TMB), o-phenylenediamine (OPD) are typical chromogenic substrates to evaluate the activity of peroxidase-like nanozymes. However, these substrates are toxic, unstable, and the coloration reactions are slow. Herein, the classical starch-sodium iodide reagent was utilized as the chromogenic substrate to evaluate peroxidase-like activity of nanozymes. Starch-sodium iodine is less toxic, more stable and faster color reaction than ABTS, TMB and OPD. Under the catalysis of peroxidase-like nanozymes such as Mo 5 N 6 nanosheets, graphene oxide nanosheets, Fe 3 O 4 nanoparticles, CeO 2 nanoparticles, iodide ion (I – ) is oxidized by H 2 O 2 to produce blue color complex (starch-I 2 ). Taking Mo 5 N 6 nanosheets as the peroxidase-like nanozyme, the color reaction completed within 5 min at room temperature (25 ºC), and the chromogenic substrate remained stable within 30 days. The maximum reaction rate (V max ) using starch-sodium iodine as the chromogenic substrate was more than 100-fold higher than that of TMB. Based on this, a sensitive colorimetric sensor and facile portable agarose gel test kits were proposed and applied for the detection of H 2 O 2 residue in liquid milk samples with fast response. • Classical chromogenic reagent starch-I 2 was utilized as the substrate of peroxidase-like nanozymes. • Starch-sodium iodine is less toxic, more stable and faster color reaction than ABTS, TMB and OPD. • A sensitive colorimetric sensor and portable agarose gel test kits were proposed for H 2 O 2 in liquid milk.