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• The fluorescence model has a ratiometric fluorescence signal to improve accuracy. • The product DAP has a yellow color can be directly observed by the naked eye. • This dual-model immunoassay shows high sensitivity and selectivity for AFB1 detection. In this work, a dual-model immunoassay for detecting Aflatoxin B1 (AFB1) was developed based on 2,3-diaminophenazine (DAP) and carbon dots (CDs). Under the catalysis of horseradish peroxidase (HRP), the o -phthalylenediamine (OPD) was oxidized to DAP which had a yellow color and intense fluorescence. The color changes form colorless to yellow was used to design absorbance model immunoassay. Meanwhile, the absorption spectrum of DAP overlapped with the emission spectrum of CDs which caused the fluorescence of CDs to be quenched. The fluorescence changes of DAP and CDs were used to develop ratiometric fluorescence immunoassay. The dual-model immunoassay showed excellent sensitivity with the limits of detection (LODs) of 0.013 ng/mL for fluorescence mode and 0.062 ng/mL for absorbance mode. Meanwhile, both models exhibited great selectivity for AFB1. Additionally, the recovery rates suggested the proposed dual-model immunoassay had great potential in actual samples detection. [ABSTRACT FROM AUTHOR]

In this work, a dual-mode immunoassay to detect Ochratoxin A (OTA) was developed relied on p -nitrophenyl phosphate (PNPP) and 2-aminoterephthalic acid (PTA-NH 2). This proposed immunoassay based on microplate immunoenzyme technique used alkaline phosphatase (ALP) as a label for forming immune complexes. The ALP was used to catalyze the dephosphorization of PNPP to generate yellow p -nitrophenol (PNP). Since the absorption wavelength of PNP overlapped with the fluorescence emission of PTA-NH 2 , the fluorescence of PTA-NH 2 can be quenched by PNP. The color of PNP and the fluorescence of PTA-NH 2 were used as double signals to detect OTA. In this dual-mode immunoassay, the limit of detection (LOD) and the linear detection range of fluorescence mode were 0.57 ng/mL (about 0.57 μg/kg) and 1.56–50.00 ng/mL (about 1.56–50.00 μg/kg), respectively. For absorbance mode, the LOD and linear range were 0.07 ng/mL (about 0.07 μg/kg) and 0.39–25.00 ng/mL (about 0.39–25.00 μg/kg), respectively. Both detection modes showed good selectivity for OTA. The recoveries rates of fluorescence mode and absorption mode in maize flours were 91.80–128.85% and 100.10–114.00%, respectively. • The dual-mode immunoassay was developed based on PNPP and PTA-NH 2. • This assay based on microplate immunoenzyme technique used ALP as a label for forming immune complexes. • The mutual verification of the two modes improved the accuracy of analysis method. • The dual-mode immunoassay had a wide linear range and low LOD. [ABSTRACT FROM AUTHOR]