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Background: The presence of aflatoxin M1, a toxic, mutagenic, and carcinogenic substance, poses significant health risks. Hence, this study assesses the levels of aflatoxin M1 in raw, pasteurized, and sterilized milk obtained from the North Khorasan Province, Iran, and evaluates its potential impact on consumer health. Methods: This descriptive-analytical study randomly collected 189 raw milk samples. Additionally, 70 pasteurized and sterilized milk samples from supermarkets in the North Khorasan Province, Iran, were included. All samples were assessed for aflatoxin M1 contamination using the enzyme-linked immunosorbent assay method. Meanwhile, statistical analysis (analysis of variance) was performed on the results. Results: The Mean±SD concentration of aflatoxin M1 in raw milk from the North Khorasan Province, Iran, was 18.44±34.6 ng/L, while pasteurized and sterilized milk had a higher mean concentration of 42.8±21.54 ng/L. The lowest concentration was found in Bojnord City, Iran, at 9.30±8.91 ng/L, while the highest concentration was detected in Jajarm at 30.70±80.50 ng/L. The mean contamination of aflatoxin M1 was not statistically significant (P=0.42) in most cities of the province. Meanwhile, 6.34% of raw milk and 11.4% of pasteurized milk samples had higher levels of aflatoxin M1 than the maximum limit recommended by the Iran National Standard Organization (INSO) (100 ng/L). However, none of the milk samples exceeded the permissible limit set by the Veterinary Organization of the country (500 ng/L). Conclusion: The mean concentration of aflatoxin M1 in raw and pasteurized milk from the North Khorasan Province is lower than the European Union (EU) standard of 50 ng/L and the approved limit set by the country’s Veterinary Organization (500 ng/L). Therefore, it is not considered a significant threat to the health of adults. [ABSTRACT FROM AUTHOR]