The aims of the present study were to investigate the occurrence of aflatoxin B 1 (AFB 1), aflatoxin B 2 (AFB 2), aflatoxin G 1 (AFG 1) and aflatoxin G 2 (AFG 2) in dairy cow feedstuff samples collected from four west provinces in Iran and estimate the corresponding concentrations of aflatoxin M 1 (AFM 1) in milk during winter and summer in 2014. A total of 160 feed samples consisting of corn silage (n = 40), alfalfa hay (n = 40), straw (n = 40) and barley (n = 40) were analyzed using high-performance liquid chromatography with a fluorescence detector method. AFB 1 , AFB 2 , AFG 1 and AFG 2 were detected in 82.5%, 69.37%, 43.12% and 41.87% of the feed samples, respectively. The concentration of AFB 1 in 65% (26/40) and 10% (4/40) of corn silage and straw samples was higher than the European Union limit, respectively. Estimation of the total corresponding concentration of AFM 1 in milk was evaluated as 20.16 and 35.68 ng/l during summer and winter, respectively.