The objective of this study was to determine the efficacy of mycotoxin sequestering agents for aflatoxin B1 (AFB1), deoxynivalenol (DON), and zearalenone (ZEA) using an in vitro method. The twelve toxin sequestering agents tested were seven bentonite products (bentonite A, B, C, D, E, F, and G), two aluminosilicate products (aluminosilicate A and B), a heulandite product, an activated charcoal product, and a yeast cell wall product. A two-step in vitro procedure was employed to mimic the conditions of temperature, pH, and digestive enzymes in the stomach and small intestine of pigs. All mycotoxin sequestering agents tested were able to bind to AFB1 with a high efficacy (>92%). The DON sequestering rate of activated charcoal (99.1%) was greater (p < 0.05) than that of other products. The ZEA sequestering rate of bentonite F (97.0%), aluminosilicate A (99.6%), and activated charcoal (100.0%) was the greatest (p < 0.05) among the tested mycotoxin sequestering agents. Overall, most mycotoxin sequestering agents had the ability to bind to AFB1, but most products, except activated charcoal, failed to sequester DON and ZEA.