BackgroundTo quantify the osmolality of human milk fortified with human milk fortifiers (HMFs), powder infant formulas, and protein additives.MethodsCommercial liquid HMFs and powder infant formulas were added to pasteurized pooled donor human milk in triplicate and stirred. The osmolality of unfortified and fortified human milk at 22, 24, 26, 27, 28, and 30 kilocalories/ounce was determined using freezing point depression.ResultsThe osmolality of fortified human milk associated with caloric density in a linear relationship regardless of the fortification strategies. Multiple liquid HMFs and every powder infant formula exceeded the osmolality threshold of 450 mOsm/kg H2O within the caloric densities tested.ConclusionThe osmolality of fortified human milk is highly variable and should be considered when selecting a fortifying agent for human milk.This article is protected by copyright. All rights reserved.