Ultrasensitive and Selective Immuno-Magnetic Ratiometric Fluorescent Sensor for Aflatoxin B1 in Food Matrices

Aflatoxin B1 (AFB1), a highly carcinogenic mycotoxin, has been the focus of research for the development of efficient detection methods. In this study, a novel magnetic immuno-ratiometric fluorescent sensing system was constructed for the quantitative detection of AFB1. Green-emitting carbon quantum dots were conjugated with AFB1 monoclonal antibody to obtain GCDs@AFB1 mAb, and AFB1 oxime was immobilized on Fe3O4 magnetic microspheres to prepare AFB1-Ox@Fe3O4 NPs. After the immune-competitive adsorption of GCDs@AFB1 mAb by AFB1-Ox@Fe3O4 NPs and free AFB1, magnetic separation was performed. Red fluorescent silver nanoclusters were introduced as an internal reference to construct a GCDs-AgNCs ratiometric fluorescent system. The sensor exhibited a good linear response in the range of 0~240 pg/mL with a low limit of detection of 18 pg/mL and excellent selectivity. The spiked recoveries in real samples ranged from 92.14% to 110.02%, with a relative standard deviation of 0.57% to 4.58%. Combining the specific antigen–antibody recognition with magnetic separation technology, this method addresses the issues of poor stability and high environmental interference of traditional fluorescent sensors, and provides a new strategy for the sensitive and stable detection of AFB1.