In vitro evaluation of binding capacity of different binders to adsorb aflatoxin

Document Type : Original Article


Department of Animal and Poultry Sciences, College of Abouraihan, University of Tehran, Iran


This study was conducted to compare the efficacy of different feed additives as mycotoxin binders in vitro. Four prevalent aflatoxin-sequestering agents (SAs) including two bentonite clays (common and acid activated bentonite), a yeast cell wall product and an activated charcoal product were evaluated in vitro to verify their capacity for binding aflatoxin B1 (AFB1). The SAs were individually mixed at two different ratios with AFB1 (1:70,000, 1:120,000) and their binding capacity indices were determined. Experimental bentonites showed high adsorption abilities, binding more than 70.00% of the available AFB1. At the 1:70,000 and 1:120,000 aflatoxin binder (AF:B) ratios, acid activated bentonite were sequestered over 87.00 and 99.00% of the AFB1, respectively. Yeast cell wall showed moderate adsorption ability at the 1:120,000 AF:B ratio, adsorbing 47.00 of AFB1. The adsorption ability of activated carbon at two AF:B ratio and yeast cell wall at 1:70,000 AF:B ratio were significantly lower than other binders. The ratio of chemisorption and binding equivalency factor were higher for acid activated bentonite compared to other sequestering agents. Based on the result of this study, it seems that acid activated bentonite could be considered efficient at sequestering the available AFB1, resulting as promising agents for use in animals diet.


  1. Alborzi S, Pourabbas B, Rashidi B, et al Aflatoxin M1 contamination in pasteurized milk in Shiraz (south of Iran). Food Control 2006; 17(7):582-584.
  2. Decastelli L, Lai J, Gramaglia M, et al. Aflatoxins occurrence in milk and feed in Northern Italy during 2004–2005. Food Control 2007; 18(10): 1263-1266.
  3. Some Naturally Occurring Substances. Available from: Accessed May 05, 2021.
  4. Diaz DE, Hagler Jr WM, Blackwelder JT, et al. Aflatoxin binders II: Reduction of aflatoxin M1 in milk by sequestering agents of cows consuming aflatoxin in feed. Mycopathologia 2004; 157(2): 233-241.
  5. Masoero F, Gallo A, Moschini M, et al. Carryover of aflatoxin from feed to milk in dairy cows with low or high somatic cell counts. Animal 2007; 9:1344-1350.
  6. Kong C, Shin SY, Kim BG. Evaluation of mycotoxin sequestering agents for aflatoxin and deoxynivalenol: an in vitro approach. SpringerPlus 2014; 3: 346. doi: 10.1186/2193-1801-3-346.
  7. Diaz DE, Hagler Jr WM, Hopkins BA, et al. Aflatoxin Binders I: In vitro binding assay for aflatoxin B1 by several potential sequestering agents. Mycopathologia 2002; 156(3): 223-226.
  8. Marroquín-Cardona A, Deng Y, Taylor JF, et al. In vitro and in vivo characterization of mycotoxin-binding additives used for animal feeds in Mexico. Food Addit Contam Part A Chem Anal Control Expo Risk Assess 2009; 26(5):733-743.
  9. Chansiripornchai P, Fink-Gremmels J, Evaluation of the aflatoxin B1 adsorption capacity of bentonite using an in vitro method mimicking monogastric gastrointestinal tract conditions. Thai J Vet Med 2004; 34: 13-19.
  10. Huwig A, Freimund S, Käppeli O, et al. Mycotoxin detoxification of animals feed by different adsorbents. Toxicol Lett 2001; 122(2): 179-188.
  11. Deng Y, Velázquez ALB, Billes F, et al. Bonding mechanisms between aflatoxin B1 and smectite. Appl Clay Sci 2010; 50(1): 92-98.
  12. Phillips TD, Lemke SL, Grant PG, Characterization of clay-based enterosorbents for the prevention of aflatoxicosis. Adv Exp Med Biol 2002; 504: 157-171.
  13. Kannewischer I, Arvide MGT, White G, et al. Smectite clays as adsorbents of aflatoxin B1: Initial steps. Clay Sci 2006; 12(Suppl. 2): 199-204.
  14. Yiannikouris A, André G, Poughon L, et al. Chemical and conformational study of the interactions involved in mycotoxin complexation with beta-D-glucans. Bio-macromolecules 2006; 7(4):1147-1155.
  15. Moschini M, Gallo A, Piva G, et al. The effects of rumen fluid on the in vitro aflatoxin binding capacity of different sequestering agents and in vivo release of the sequestered toxin. Anim Feed Sci Technol 2008; 147(4):292-309.
  16. Grant PG, Phillips TD. Isothermal adsorption of aflatoxin B(1) on HSCAS clay. J Agric Food Chem 1998; 46(2): 599-605.
  17. Magnoli AP, Alonso VA, Cavaglieri LR, et al. Effect of monogastric and ruminant gastrointestinal conditions on in vitro aflatoxin B1 adsorption ability by a montmorillonite. Food Add Cont Part A Chem Anal Control Expo Risk Assess 2013; 30:743-749.
  18. Ledoux DR, Rottinghaus GE. In vitro and in vivo testing of adsorbents for detoxifying mycotoxins in contaminated feedstuffs. In: Lyons TP, Jacques KA (Eds). Biotechnology in the Feed Industry. Nottingham, UK: Nottingham University Press, 1999; 369-379.
  19. Vekiru E, Fruhauf S, Sahin M, et al. Investigation of various adsorbents for their ability to bind aflatoxin B1. Mycotoxin Res 2007; 23(1):27-33.
  20. Gallo A, Masoero F. In vitro models to evaluate the capacity of different sequestering agents to adsorb aflatoxins. Ital J Anim Sci 2010; 9: e21. doi:10.4081/ ijas.2010.e21.
  21. Desheng Q, Fan L, Yanhu Y, et al. Adsorption of aflatoxin B1 on montmorillonite. Poultry Sci 2005; 84(6): 959-961.
  22. Komadel P. Chemically modified smectites. Clay Miner 2003; 38(1):127-138.