Objectives: to verify the effect of three storage conditions on human milk acidity and to determine the quantity of lipids, total solids, ash, calcium, and total proteins. Methods: the sample comprised healthy milk donors to the Human Milk Bank of the University Hospital of Maringá (n37), State of Paraná, Brazil. The quantities of lipids, protein, total solids, ash and calcium were measured, as well as acidity. The study protocol included three storage conditions: time zero, after 4h storage at room temperature and after 24h under refrigeration. Results: the average concentrations of nutrients found in this study were comparable to those described in milk from lactating mothers of developed countries or other Brazilian regions, which is suitable forfeeding newborns. As for acidity, variations were observed between individuals for the storage condi- tions involving 4h and 24h, which did not occur for the condition of Oh. A weak correlation between the variables of acidity and protein content was observed, as well as for acidity and calcium. Conclusions: the acidity does not have a sign iji- cant influence on the concentration of the nutrients, at least under the storage conditions studied here. The best way to preserve human milk is to freeze it imme- diately after collection. [ABSTRACT FROM AUTHOR]