Magnetic-encoded fluorescent nanospheres-based competitive immunoassay for near-simultaneous detection of four...

Simultaneous detection of mycotoxins is important for food safety. In this study, a magnetic-encoded fluorescent nanosphere-based competitive immunosensor (cFMEIS) with 2×2 array was first developed for simultaneous detection of aflatoxin B₁ (AFB₁), ochratoxin (OTA), deoxynivalenol (DON), and zearalenone (ZEN) in wheat. Specifically, magnetic nanoparticles with strong and weak responses were conjugated with mycotoxin antigens as capture probes. Fluorescent nanoparticles doped with europium ion (Eu³⁺) and terbium ion (Tb³⁺) with red and green emission were coupled with mycotoxin antibodies as signal probes. Using a magnetic field, immune complexes were sequentially separated in a complex system and fluorescently detected. The detection limits of AFB₁, DON, OTA, and ZEN were 0.032, 0.141, 0.097, and 0.376 μg/kg, respectively. The recoveries in the certified reference material of wheat flour ranged from 81.6 to 120.0 %. Owing to its high accuracy, selectivity, and sensitivity, the cFMEIS shows great promise as an efficient and sensitive multitarget sensor for mycotoxins.