The objective of the study was to evaluate the capacity of 6 mycotoxin binders to adsorb 2 groups of vitamins: water-soluble vitamins (WSV) and fat-soluble vitamins (FSV). The experiment was conducted in in vitro condition to simulate post ruminal digestion model in 2 phases: the first phase to simulate the gastric digestion with pepsin, malic acid, citric acid, acetic acid and lactic acid at pH 3.0; and the second phase to simulate intestinal digestion with bile salts and pancreatin extract at pH 6.5. The experimental design was a factorial 6 × 5 with main factors being mycotoxin binders (bentonite, clinoptiolite, sepiolite, montmorillonite, active carbon and yeast cell wall) and vitamins (WSV: B2, B3, B6; and FSV: D and E). Vitamins were incubated separately (Study 1) and together (Study 2) with each mycotoxin binder. Each treatment was incubated in triplicate and 2 consecutive periods. Results were analyzed with PROC MIXED of SAS and significance declared at P < 0.05. In the 2 studies the adsorption average was highest with montmorillonite for both WSV (47%) and FSV (27%) and lowest with active carbon (17%) for WSV and sepiolite (12%) for FSV. For the adsorption average of vitamins in the 2 studies, the B6 (55%), B2 (33%) and E (35%) were adsorbed the highest, and B3 (5%) and D (3%) were adsorbed the lowest. The adsorption of vitamin B6, B2 and E was highest with montmorillonite (68%, 67% and 48%, respectively), and lowest with clinoptiolite (50%) for B6, with active carbon (4%) for B2 and with sepiolite (15%) for E in both studies. For vitamin D, the adsorption was highest with yeast cell wall (20%) and lowest with bentonite (0%) in Study 1; however, in Study 2, the adsorption for vitamin D was highest with montmorillonite (7%) and lowest for all other adsorbents (0%). Mycotoxin binders may affect bioavailability of some water- and fat- soluble vitamins at different degree depending on the mycotoxin binders and the substrate.