Nitrofurans such as furaltadone and nitrofurantoin are a type of synthetic broad-spectrum antibiotics. Various fluorescent nanomaterials have been used as labeling materials in immunochromatographic assays (ICAs) for nitrofurans detection. However, previous fluorescent nanomaterials can undergo aggregation-caused quenching, leading to a decrease in the detection sensitivity. In this study, we developed a multiplex immunochromatographic assay (mICA) based on dual-color aggregation-induced emission nanoparticles (AIENPs) as signal labels for the simultaneous detection of 3-amino-5-morpholino-methyl-1,3-oxazolidinone (AMOZ) and 1-aminohydantoin (AHD), which were the metabolites of furaltadone and nitrofurantoin, respectively. Under optimal conditions, the cut-off values of the mICA for derivatized AMOZ (2-NP-AMOZ) and AHD (2-NP-AHD) reached up to 3 and 5 ng/mL, respectively. These values are at least 166-and 200-fold higher than those of the commercial gold nanoparticles (GNPs)-based test strip, respectively. Furthermore, the test strip was successfully applied to the samples, with acceptable recoveries in the range of 83.0–98.2%.
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