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Introduction: Breast milk is essential for human development, but it may contain toxics derived from environmental pollution, especially in mining areas. Objective: To assess the prevalence of mercury contamination in breast milk and factors associated with its transfer to nursing mothers living in municipalities with gold mining. Materials and methods: We conducted a cross-sectional study with 150 nursing mothers in four municipalities of Antioquia (El Bagre, Segovia, Remedios and Zaragoza) with a mining tradition. We surveyed these mothers to obtain information on their sociodemographic, occupational and environmental factors related to mercury, and we took breast milk, urine and hair samples. We calculated mercury level averages and the contamination prevalence per municipality. Results: Mercury average in breast milk was 2.5 (± standard deviation: 9.2) mg/L. The prevalence of mercury contamination in samples with a high level of mercury was 11.7%. Conclusion: This study shows that there is a serious problem of mercury contamination in gold mining regions of Antioquia, with significant involvement of the most vulnerable population. [ABSTRACT FROM AUTHOR]