A nanozyme catalysis-diazotization reaction cascaded ratiometric colorimetric assay probing the dynamics of nitrite in leftovers

Ziyu Zhang, Lina Tang, Wenjie Sheng, Youyi Yang, Guolin Lai, Jinjin Liu, Xiangheng Niu
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Abstract

As a common food additive, excessive nitrite poses a great threat to human health, and monitoring its content in food products is of importance for healthy diet. Currently, the detection of nitrite content in food is primarily focused on fresh dishes, and there is a lack of research on monitoring the variations of nitrite in different foods under various storage conditions. In this study, we cascaded nanozyme catalysis with diazotization reaction and developed a ratiometric colorimetric assay to dynamically analyze nitrite in leftovers. First, nanoscale MnFe2O4 was synthesized as an oxidase mimic to catalyze colorless 3,3,5,5-tetramethylbenzidine (TMB) oxidation to blue TMBox. Then, a diazotization process of the produced TMBox took place under the stimulation of nitrite, lowering the ultraviolet-visible absorption signal (652 nm) assigned to TMBox and simultaneously generating a new signal at 445 nm ascribed to diazotized TMBox. Thus, a ratiometric colorimetric method could be constructed based on the above reversed variations of the two signals for high-selectivity nitrite determination, providing a linear range of 1.76–180 μM and a detection limit of 0.12 μM. By employing the established assay to dynamically monitor nitrite in food products, it was found that the nitrite content in overnight leftovers was higher than that in fresh dishes, presenting an increasing trend with storage time. In addition, potential impacts of storage factors on the dynamics of nitrite content were investigated, providing some implications for food preservation and daily consumption.

Abstract Image

纳米酶催化-重氮化反应级联比例比色法测定剩饭剩菜中亚硝酸盐的动力学
亚硝酸盐作为一种常见的食品添加剂,过量的亚硝酸盐对人体健康构成了极大的威胁,监测其在食品中的含量对健康饮食具有重要意义。目前,对食品中亚硝酸盐含量的检测主要集中在新鲜菜肴上,缺乏对不同食品在不同储存条件下亚硝酸盐含量变化的监测研究。在这项研究中,我们将纳米酶催化与重氮化反应级联,并建立了一种比色法来动态分析剩饭剩菜中的亚硝酸盐。首先,合成纳米级MnFe2O4作为氧化酶模拟物,催化无色3,3 ',5,5 ' -四甲基联苯胺(TMB)氧化生成蓝色TMBox。然后,生成的TMBox在亚硝酸盐的刺激下发生重氮化过程,降低了分配给TMBox的紫外可见吸收信号(652nm),同时在重氮化TMBox的445nm处产生了新的信号。因此,基于上述两种信号的反向变化,可以构建高选择性亚硝酸盐测定的比例比色法,线性范围为1.76 ~ 180 μM,检出限为0.12 μM。利用所建立的方法动态监测食品中亚硝酸盐的含量,发现隔夜剩菜中的亚硝酸盐含量高于新鲜菜中的亚硝酸盐含量,并随着存放时间的延长呈增加趋势。此外,还研究了贮藏因素对亚硝酸盐含量动态变化的潜在影响,为食品保鲜和日常食用提供参考。
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