Breastfeeding plays an important role in the growth and development of breastfed infants, especially in the first 6 months of their lives. The present study was conducted to determine the nickel concentrations in breast milk of lactating women in Yazd, Iran. One hundred fifty volunteers were selected among nursing mothers referring to health centers in Yazd. In the first month of lactation, milk samples were collected three times, on days 3 to 5 (first), 16 (Second), and 30 (third) after delivery. Nickel concentration of the samples was measured by atomic absorption spectrophotometer. Demographic variables were collected through a questionnaire which was completed by mothers. The mean age of the study group was 27.40 ± 4.66 years. The mean nickel concentrations in breast milk at the first, second, and third samples were 47.3 ± 7.40, 49.9 ± 8.05, and 54.8 ± 7.38 μg/l, respectively. The concentration of nickel in the breast milk of more than 86 % of mothers was higher than the permissible range for it. There was no significant relationship between the mean value of nickel in breast milk and education, age, and job of mothers. High level of nickel in breast milk may be attributed to consumed food and drinking water containing nickel. Monitoring the nickel level in breast milk regularly is recommended.