Aflatoxins which are highly toxic, immunosuppressive and carcinogenic secondary metabolites produced naturally by Aspergillus flavus fungal species have a harm effect on human and animal health. Label free and rapid sensing of aflatoxin B1 (AFB1) has drawn the increased interest of highly sensitive and selective research. A highly sensitive and selective plasmonic sensing method was developed for the detection of AFB1 based on enhance-surface plasmon resonance nanosensor. Firstly, AFB1 and N-methacryloyl-l-phenylalanine were pre-complexed as a template molecule and functional monomer. Molecularly imprinted polymers with gold nanoparticles were coated onto surface plasmon resonance (SPR) gold chip surface. The AFB1 imprinted nanosensor shown a wide linear range, between 0.0001 ng mL−1 and 10.0 ng mL−1, and the limit of detection is 1.04 pg mL−1. Compared to the non-imprinted nanosensor, the imprinting factor was found to be 5.91. Also, detection studies of AFB1 were performed using various food samples. Finally, SPR nanosensors were performed selectivity, reusability and storage stability analysis.