Fe-Ni dual-single atoms nanozyme with high peroxidase-like activity for sensitive colorimetric and fluorometric dual-mode detection of cholesterol

Cholesterol is widely existed in nerve myelin sheath and various membrane structures, whose abnormal level would deteriorate human cells or even cause diseases. Herein, Fe-Ni dual-single-atom nanozyme was efficiently incorporated into N-doped carbon nanosheets (FeNi DSAs/N-CSs) by a simple calcination method. Its nanozyme activity and catalytic mechanism were investigated in details. The FeNi DSAs/N-CSs nanozyme showed superior peroxidase-like property, which was applied for the dual-mode determination of hydrogen peroxide (H₂O₂) and cholesterol. The colorimetric/fluorometric assays of H₂O₂ displayed the linear ranges of 1–50 mM and 5–40 mM with low limits of detection of 0.45 mM and 3.33 mM, respectively. In parallel, there exhibited the linear ranges of 0.5–5.0 mM and 0.25–5.0 mM for the colorimetric/fluorometric analysis of cholesterol, coupled with the limits of detection down to 0.19 mM and 0.044 mM, respectively. This work provided a rapid, cost-effectiveness and simple colorimetric/fluorometric method for sensitive dual-mode detection of cholesterol in human serum samples.