Experiments were carried out to evaluate the effectiveness of three different biosorbents (banana peel, leaves, and powder) in removing aflatoxin B₁ (AFB₁). A noncommercial mycotoxin binder (zeolite) was used as a reference material. A laboratory model that simulated the in vivo conditions of the poultry gastrointestinal tract was utilized to prove the removal efficiency of the biosorbents when added to AFB₁-contaminated diet (100 µg/kg). The concentration of AFB₁ was determined using antibody-based immunoaffinity column and spectrofluorometry methodologies. Z potential (ζ), point of zero charge (pH), energy-dispersive X-ray spectroscopy (EDS), X-ray diffraction (XRD), Fourier transform infrared spectroscopy with attenuated total reflection (FTIR-ATR), and UV-Vis diffuse reflectance spectroscopy (DRS) techniques were used to further characterize the biosorbents. The addition of the biosorbents (1.5%, /) to the diet significantly reduced the bioavailability of AFB₁ in the intestinal section. The highest aflatoxin adsorption values were 69% and 70% using powder and zeolite, respectively. A moderate biosorption uptake of 46% was achieved using leaves. The biomaterial with the lowest removal capacity was banana peel (28%). In conclusion, powder could be used as an alternative to conventional systems for AFB₁ removal.