We report an aptamer-based biosensor for detection of aflatoxin B1 (AFB1), a mycotoxin identified as contaminant in food. The sensor is assembled in a multilayer framework that utilizes cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) for acquiring the signal response by means of redox indicators: K[Fe(CN)6]-3/-4. Poly (amidoamine) dendrimers of fourth generation (PAMAM G4) immobilized on gold electrode covered by cystamine, were employed for attachment of single stranded amino-modified DNA aptamers specific to AFB1. The cystamine-dendrimers (Cys-PAMAM) layers were compared with other immobilization platforms such as cystamine (Cys), 11-mercaptoundecanoic acid (MUA) and 11-mercaptoundecanoic acid-dendrimers (MUA-PAMAM), being the first approach the most appropriate for producing sensitive and reproducible signal in the range of concentrations 0.1-10nM AFB1. The sensor was validated in certified contaminated peanuts extract as well as in spiked samples of peanuts-corn snacks and the sensing response was evaluated and compared in terms of the matrix effect. The aptamer specificity was analyzed by testing the sensor in other mycotoxins such as aflatoxin B2 (AFB2) and ochratoxin A (OTA). The limit of detection achieved by this sensor was LOD=0.40±0.03nM, it was regenerable in 0.2M glycine-HCl and it did not lose its stability up to 60h storing at 4°C. Atomic Force Microscopy (AFM) studies were also performed for illustrating individual steps of biosensor assembly.

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