Adapted, follow-up, probiotic follow-up, toddler, and probiotic toddler infant formulas were subjectedto an in vitro enzymatic procedure simulating physiological digestion. The formation and identificationof casein phosphopeptides (CPPs) in the milk-based infant formulas were studied using reversedphase high-performance liquid chromatography coupled on line to an ion trap mass spectrometer.Most CPPs formed contained the cluster sequence SpSpSpEE, a mineral binding site. Phosphopeptide
s2-CN(1-19)4P was present in all formulas analyzed. Probiotic formulas released CPPs not detectedin nonprobiotic formulas and probably formed by bifidobacteria action. These observations suggestthat physiological digestion of these products promotes the formation of bioactive peptides with mineralcarrier properties in the gastrointestinal tract, which resist further proteolysis.Keywords: Casein phosphopeptides; simulated digestion; mineral bioavailability; milk-based infantformulas; reversed phase high-performance liquid chromatography; mass spectrometry