Analysis of peptides from bovine hemoglobin and tuna myoglobin enzymatic hydrolysate: use of HPLC with on-line second-order derivative spectroscopy for the characterization of biologically active peptides

The Chromatographic resolution of bovine hemoglobin and tuna myoglobin peptic hydrolysates was performed by use of successive SE-and RP-HPLC in order to obtain pure peptides. Analysis of their amino acid composition and second order derivative spectroscopy allowed us to determine the accurate identity of these peptides and consequently to locate them in both protein sequences. Some biologically active peptides generated from hemoglobin were also studied by these methods to investigate the kinetics of their appearance during peptic hydrolysis.