Alaska pollock skin as aquatic by-product was usually discarded in food processing. Iron deficiency has been a nutritional problem for a long period. In this study, Alaska pollock skin was used to generate iron-chelating peptides after being treated by commercial enzymes. Sequential chromatography, including immobilized metal affinity chromatography (IMAC) and reversed phase high performance liquid chromatography (RP-HPLC), was used for capturing and purifying iron-chelating peptides. One tripeptide, with high iron-chelating activity, was obtained and identified by electrospray ionization tandem mass spectrometry (LC ESI-MS/MS). The amino acid sequence of the iron-chelating peptide was identified to be Ser-Cys-His (MW: 345 Da). These results show that Alaska pollock skin could be utilized to generate iron-chelating peptides as an iron supplement in functional food industry. (C) 2013 Elsevier Ltd. All rights reserved.