Objective. To determine the recommended dose of bentonite as a chelator of arsenic (As) and lead (Pb) in the rumen, abomasal and duodenal environment by means of an in vitro system, considering the variables of ruminal fermentation and the adsorbent effect of As and Pb. Materials and Methods. Vials were prepared with 50 mL of anaerobic medium and 0.5 g of alfalfa as substrate. The culture media were added with 1.01 and 1.95 ppm of As and Pb, respectively. The treatments consisted of different levels of bentonite (B) added to the medium, B-0%, B-1.5%, B-3.0% and B-4.5%. The media were incubated for 24 h, and pH, volatile fatty acid (VFA) and N-NH3 production, in vitro DM digestibility (IVDMD) and chelating effect of bentonite were measured, simulating rumen, abomasum and duodenum conditions. A completely randomised design and orthogonal contrasts were used. Results. Bentonite did not modify the pH (p > 0.05) and the molar proportion of VFAs, but the total VFA concentration presented a negative linear effect with increasing bentonite levels. Likewise, the N-NH3 level decreased (p < 0.05) with increasing inclusion levels of bentonite. The IVDMD was reduced with B-3% and B-4.5% (52.36b and 49.74b%) in comparison to B-0% and B-1.5% (62.30a and 61.98a%). Treatments B-1.5% and B-4.5% improved (p < 0.05) As and Pb adsorption in the rumen, abomasal and duodenal environments. Conclusions. The recommended level of bentonite as chelator of As and Pb is 1.5%; doses of 4.5% affect IVDMD and, consequently, animal productivity.