An antioxidative peptide was isolated from the pellet by first dissolving the pellet in DMSO/80 % (v/v) aqueous ethanol (1/1, v/v) and then loaded onto a cation-exchange resin column. The eluate was monitored at 215 nm to trace peptide content, and antioxidative activity indicated by the A593 was then measured by FRAP assay for each fraction (5 ml).Figure 1a shows a typical elution profile, indicating that the major antioxidative activity (fractions 12–22 in this example) was eluted with 100 mM NaCl. Next,the fractions with high antioxidant activity were pooled and applied to preparative RP-FPLC eluted with a linear gradient of aqueous acetonitrile (0–80 % by volume). Figure 1b shows a representative RPFPLC elution curve. FRAP assay results revealed four antioxidative peaks, assigned as Fa-1, Fa-2, Fa-3, and Fa-4. Fraction Fa-1 was applied to RF-HPLC again,and the resulting fractions possessing antioxidative activity were pooled and used for amino acid sequence
identification.