The objectives of this study were to observe the extent of transfer of aflatoxin B1 in feed to the aflatoxin M1 metabolite in milk in Nili-Ravi buffaloes and to evaluate the efficacy of a commercial mycotoxin binder (Mycofix, Biomin Singapore) incorporated into feed to minimise this transfer. Multiparous animals (n ≤ 28) were randomly distributed to four groups corresponding to two treatments each with two levels of aflatoxin B1. Individual animals were exposed to naturally contaminated feed providing a total of 1475 μg/day (Groups A and B) or 2950 μg/day (Groups C and D) of aflatoxin B1. Groups B and D were given 50 g of mycotoxin binder daily mixed with feed whereas Groups A and C were kept as controls. Feed samples were analysed by reverse phase high performance liquid chromatography for aflatoxin B1 and milk samples were evaluated by enzyme-linked immunosorbent assay for the liver metabolite aflatoxin M1. The mean value of total daily aflatoxin M1 excretion for animals fed 2950 μg/day of aflatoxin B1 (112.6 μg/day) was almost double (P < 0.001) than the excretion in buffaloes fed 1475 μg/day (62.2 μg/day). The mean daily concentration of aflatoxin M1 in milk of animals from both treatment groups supplemented with 50 g/day of mycotoxin binder was 76.5 μg/day, nearly 22 μg lower than those without binder at 98.3 μg/day (s.e.d. ≤ 5.99: P < 0.01). The interaction of binder and treatment was not significant i.e. the 50 g/day of binder was able to sequester aflatoxin B1 with the same efficiency in groups fed with high and low concentrations of aflatoxin B1. Carry over was (3.44%) lower (P ≤ 0.001) in animals supplemented with 50 g/day of mycotoxin binder than those fed no binder (4.60%). Thus buffaloes are highly efficient at transferring aflatoxins in feed to the aflatoxin M1 metabolite in milk, whereas mycotoxin binder is capable of alleviating without preventing this contamination risk.