Separation of a casein hydrolyzate by HPSEC with a new mobile phase and characterization of peptides by FABMS

A mobile phase which was firstly developed for the separation of a peptic hydrolysate from bovine blood haemoglobin by size-exclusion chromatography (SEC) has been modified for its application to a casein hydrolysate phosphorylated by optimizing nonideal size-exclusion behavior. Peptidic fractions obtained by high performance size-exclusion chromatography (HPSEC) were further separated by reversed-phase HPLC (RP-HPLC) and characterized by fast atom bombardment mass spectrometry (FAB-MS) in order to ascribe mass and sequence (tandem-MS) to peptides. Results obtained confirmed that this HPSEC process involves mostly ion exchange and size-exclusion. This HPSEC Chromatographic procedure was found to be suitable for comparison of casein hydrolysates during production.