Portable and point-of-care visual detection of nitrite in food samples using a dual-output oxidase-mimicking ag-MOF nanozyme with ratiometric colorimetric and tonality-based response
Point-of-care testing demands ultra-sensitive, real-time, and rapid detection of nitrite to address critical challenges in... See more
Point-of-care testing demands ultra-sensitive, real-time, and rapid detection of nitrite to address critical challenges in environmental monitoring, food safety assurance, and on-site health diagnostics, owing to the widespread presence and high toxicity of nitrite across ecological and biological systems. In this study, a novel and stable silver-based metal-organic framework (Ag-MOF) was synthesized and utilized for the selective detection of nitrite. The fabricated Ag-MOF functioned as a potent oxidase mimic, catalyzing the transformation of 3,3′,5,5′-tetramethylbenzidine (TMB) into its oxidized derivative (oxTMB+.), which manifested visually as a vivid blue coloration. Upon the introduction of nitrite ions, a diazotization reaction was triggered, converting oxTMB+. into its dicationic form (oxTMB2+), resulting in a distinct color transition from blue to green-yellow. Capitalizing on this visually perceptible shift, a dual-mode detection strategy was successfully implemented, enabling nitrite quantification through two distinct approaches: (i) smartphone-assisted visual and RGB-based analysis, and (ii) conventional instrument-based fluorescence measurements using a spectrofluorometer. The oxidase-mimicking Ag-MOF exhibited a Michaelis–Menten constant (Km) of 33.4 μM, reflecting its high affinity for the substrate and enhanced catalytic efficiency, particularly at low substrate concentrations. Notably, this nanozyme is both cold- and heat-adapted, maintaining its oxidase-like activity across a broad temperature range from 5 °C to 100 °C without any loss of performance. To enable precise detection, we designed a ratiometric colorimetric platform that allows both conventional and smartphone-based analysis, utilizing color tonality changes for quantification. The traditional colorimetric approach exhibited a linear detection range for nitrite from 0.0 to 66.0 μM, achieving a low detection limit of 0.011 μM. In comparison, the smartphone-integrated method demonstrated linearity between 0.0 and 60.0 μM, with a corresponding detection limit of 0.12 μM. The Ag-MOF platform enabled smartphone-based nitrite detection in water, showing practicality, sensitivity, and promise for rapid environmental monitoring.
2025-09