AIM: To analyse the dynamics of macronutrients in breast milk in Slovakian women and compare the dynamics between mothers of hospitalized newborns and donors of human breast milk relative to the gestational age of their babies. METHODS: Human milk samples were collected from 101 breastfeeding women and analysed once or repeatedly from September 2017 to August 2018. Group A consisted of 79 mothers of newborns hospitalized at the Neonatal Department of Intensive Medicine (NDIM) of whom 69 were mothers of premature newborns (subgroup A1) and 10 were mothers of term babies (subgroup A2). Group B consisted of 22 breast milk donors registered at the Human Milk Bank at National Institute of Children's Diseases (NICHD). Of these, 4 were mothers of premature newborns (subgroup B1) and 18 were mothers of term newborns (subgroup B2). From subgroup A1, we chose two mothers, one with a hypotrophic newborn and another with a eutrophic newborn. The results were obtained by using the MIRIS breast-milk analyzer. RESULTS: The overall dynamics of macronutrients correspond with other studies, however, we demonstrated that the level of macronutrients in individual cases were different. CONCLUSION: The determination of macronutrients in human milk is essential in neonatal care. It provides information about the nutritional value of breast milk and helps to optimise nutrition according to the individual needs of newborns (Fig. 10, Ref. 16). Text in PDF www.elis.sk.