Ratiometric fluorescence and absorbance dual-model immunoassay based on 2,3-diaminophenazine and carbon dots for detecting Aflatoxin B1

Abstract

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.

Keywords: Aflatoxin B1; Carbon dots; Dual-model; Immunoassay; Ratiometric fluorescence.