This study developed an electrochemical immunosensor for the detection of aflatoxin B1 (AFB1) in vegetable oil, based on an electrochemical modified carbon cloth (EMCC) electrode modified with a composite functional layer of cross-linked o-aminothiophenol functionalized AuNPs (o-ATP@AuNPs)/Prussian Blue (PB). The EMCC electrode substrate was prepared by modifying carbon cloth through electrochemical methods to increase its surface area, which allowed for the effective deposition of o-ATP@AuNPs/PB composite functional layer and improved the conductivity of the electrode material. The synergistic effect of o-ATP@AuNPs and PB significantly enhanced the sensitivity of the electrochemical sensor. Additionally, the AuS bond between L-Cysteine (L-Cys) and o-ATP@AuNPs improved the stability of the sensing interface. Under optimal conditions, the BSA/anti-AFB1/L-Cys/o-ATP@AuNPs/PB/EMCC sensor was able to detect AFB1 in the range of 0 to 20 ng mL using square wave voltammetry (SWV), with a detection limit of 0.015 ng mL. The proposed sensor holds promise for future applications in the sensitive detection of AFB1 in vegetable oils.
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