Proteins from sunflower seeds were hydrolyzed with Alcalase and Flavourzyme to isolate iron-binding peptides. The optimal hydrolysis conditions were determined. Hydrolysates were filtered under a 3 kDa membrane and iron-binding peptides separated from the hydrolysates using ion exchange and gel permeation chromatographic methods. A fraction with the highest iron-binding activity (Fe/peptide, 0.69), F22, was obtained. These results suggest that fractions isolated from sunflower seed protein hydrolysates can be applied toward the production of iron supplements.