Peptidomic analysis of antioxidant peptides from porcine liver hydrolysates using SWATH-MS

There is a growing interest in the production and identification of bioactive peptides as health-promoting agents. A relevant method to produce biopeptides is enzymatic hydrolysis from protein-rich meat by-products. Pork liver proved to be a good source of protein (18.54%) with a low-fat content (3.38%). After hydrolysis at different times (4,6,8 and 10 h) with Alcalase, relevant amino acids such as hydrophobic (leucine, valine and isoleucine) and aromatic (tyrosine and phenylalanine) involved in antioxidant capacity were strongly increased. For the peptidomic analysis, a novel technique called sequential window acquisition of all theoretical mass spectra (SWATH-MS) was used. Regarding the effect of hydrolysis time, PCA demonstrated a great differentiation among the peptidomic pattern. Fifty-one peptides were correlated with antioxidant activity measured by DPPH, ABTS, FRAP and ORAC assays. SWATH-MS allowed the identification and quantification of six peptides from trypsinogen, ferritin, keratin, carboxylic ester hydrolase and globin domain-containing protein as potential antioxidant compounds. SIGNIFICANCE: The pork liver tissue contains a substantial amount of proteins whose enzymatic hydrolysis might generate antioxidant peptides. The bioactive peptides from pork liver would contribute to harnessing by-products of the swine industry as well as added-value products will be produced. The antioxidant activity of the mixtures revealed potential antioxidant peptides which could be used in the development of nutraceutical and functional food products.