Ratiometric Colorimetric Detection of Nitrite Realized by Stringing Nanozyme Catalysis and Diazotization Together

Due to the great threat posed by excessive nitrite in food and drinking water to human health, it calls for developing reliable, convenient, and low-cost methods for nitrite detection. Herein, we string nanozyme catalysis and diazotization together and develop a ratiometric colorimetric approach for sensing nitrite in food. First, hollow MnFeO (a mixture of Mn and Fe oxides with different oxidation states) derived from a Mn-Fe Prussian blue analogue is explored as an oxidase mimic with high efficiency in catalyzing the colorless 3,3',5,5'-tetramethylbenzidine (TMB) oxidation to blue TMBox, presenting a notable signal at 652 nm. Then, nitrite is able to trigger the diazotization of the product TMBox, not only decreasing the signal at 652 nm but also producing a new signal at 445 nm. Thus, the analyte-induced reverse changes of the two signals enable us to establish a ratiometric colorimetric assay for nitrite analysis. According to the above strategy, facile determination of nitrite in the range of 3.3-133.3 μM with good specificity was realized, providing a detection limit down to 0.2 μM. Compared with conventional single-signal analysis, our dual-signal ratiometric colorimetric mode was demonstrated to offer higher sensitivity, a lower detection limit, and better anti-interference ability against external detection environments. Practical applications of the approach in examining nitrite in food matrices were also verified.