Extractability of antioxidants from legume seed flour after cooking and in vitro gastrointestinal digestion in comparison with methanolic extraction of the unprocessed flour

Antioxidant activities were studied in methanolic and water extracts of nonprocessed, cooked and in vitro enzymatically digested seed flour, as well as in total protein hydrolysates and small peptide fractions ( lt 3 and lt 10kDa) of three pea and five grass pea cultivars. The antioxidative properties were determined by three spectrophotometric methods: 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, Folin-Ciocalteu (FC) reducing capacity assay and H2O2 scavenging. We also applied one luminometric assay for hydroxyl radical scavenging. The study showed that cooking and enzymatic digestion strongly enhanced the release of phenolic compounds in methanolic extracts of four analysed cultivars. Scavenging activity against DPPH radical, hydroxyl radical and hydrogen peroxide was increased in majority of analysed flour specimens subjected to processing. Our findings indicate that, besides the phenolic compounds, the small peptide fraction, especially the MW lt 3kDa, in methanolic and aqueous extracts of cooked and digested seed flour significantly contribute to free radical and hydrogen peroxide scavenging activity in all investigated cultivars. Our data strongly suggest that simple cooking treatment and in vitro digestion of seed flour applied prior to extraction with methanol could improve antioxidative activity of obtained extracts