An integrated aflatoxin B1 (AFB1) detection platform with quantum dot (QD)-based electrochemical immunosensor and an automated magneto-controlled pretreatment system was successfully developed. The automated pretreatment system adopts the immunoaffinity magnetic beads (IMB) as the capture probe of AFB1 and QD-labeled AFB1 complete antigen (AFB1-BSA-QDs) as the signal probe. AFB1-BSA-QDs can be easily converted into corresponding metallic cations through acidic treatment, which can be detected electrochemically via anode stripping voltammetry (ASV). Moreover, a disposable screen-printed electrode (SPE) without requiring any further modification is used in the novel electrochemical immunosensor' making routine testing feasible. Under optimal conditions, the detectable concentration range of AFB1 was 0.08-800 μg/kg. The metal ion signal associated linearly with the logarithm of AFB1 concentration within the range of 5-240 μg/kg, with a detection limit of 0.05 μg/kg. The spiked recoveries of three different concentrations in four different matrixes ranged from 83.9 to 118.0%, and inter-day relative standard deviations were below 10%. Furthermore, the methodology was validated by analyzing naturally contaminated samples, and results of the novel immunosensor were in good agreement with those of LC-MS/MS, demonstrating the potentiality of the developed method for the monitor of AFB1 in cereals and oils.