Novel Immunosensor Based on Metal Single-Atom Nanozymes with Enhanced Oxidase-Like Activity for Capsaicin Analysis in Spicy Food

Capsaicin (CAP) is a primary indicator for assessing the level of pungency. Herein, iron-based single-atom nanozymes (SAzymes) (Fe/NC) with exceptional oxidase-like activity were used to construct an immunosensor for CAP analysis. Fe/NC could imitate oxidase actions by transforming O2 to •O2– radicals in the absence of hydrogen peroxide (H2O2), which could avoid complex operations and unstable results. By regulating the Fe atom loads, an optimal Fe0.7/NC atom usage rate could improve the catalytic activity (Michaelis–Menten constant (Km) = 0.09 mM). Fe0.7/NC was integrated with goat antimouse IgG by facile mix incubation to develop a competitive enzyme-linked immunosorbent assay (ELISA). Our Fe0.7/NC immunosensing platform is anticipated to outperform the conventional ELISA in terms of stability and shelf life. The proposed immunosensor provided color responses across 0.01–1000 ng/mL CAP concentrations, with a detection limit of 0.046 ng/mL. Fe/NC may have potential as nanozymes for CAP detection in spicy foods, with promising applications in food biosensing.