Important role of milk in human, especially in infant nutrition is well known. Aflatoxins are highly toxic, mutagenic, teratogenic and carcinogenic compounds. Aflatoxin M1 is an important mycotoxin frequently found in milk and dairy products. The aim of this study was to evaluate Aflatoxin M1 contamination in raw milk samples in Mashhad, northeast of Iran by ELISA. For this purpose two hundred raw milk samples were obtained from dairy plants of Mashhad city of Iran. AFM1 was found in 76% of the raw milk samples by average concentration of 0.061 ± 0.004 µg/kg. The range of contamination levels varied among different months.The highest mean concentration of aflatoxin M1 (AFM1) in raw milk samples was registered in February samples (0.225 µg/kg) and the lowest mean concentration of aflatoxin M1 was registered in August samples (0.0058 µg/kg). Statistical evaluation showed that there were significant differences (P<0.05) between the concentrations of AFM1 of raw milk samples taken in May and August with November and February. The concentration of AFM1 in 36.5% of the samples were higher than maximum tolerance limit accepted by European union/Codex Alimentarius Commission (0.05 µg/kg). It was therefore concluded that, the levels of AFM1 in milk samples consumed in Mashhad, Iran were high and appear to be a serious public health problem at the moment. To achieve a low level of AFM1 in milk, cow\\\\\\\'s feed samples from various cowsheds must be evaluated routinely for aflatoxin and kept away from fungal contamination as much as possible.

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