Antioxidant Activities of Steamed Extract from Squid (Todarodes pacificus) Muscle

The purpose of this study was to purify antioxidant substances from steamed squid extract (SSE). The yield of SSE was 8% by dry weight. The approximate compositions of SSE proteins, lipids, moisture, carbohydrate and ash were 64.95%, 1.69%, 7.23%, 4.44% and 21.69%, respectively. The major amino acids in SSE were taurine (29.17%), glycine (20.33%), alanine (12.51%), and glutamic acid (9.83%). Antioxidant activities were evaluated using 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging activity, which was measured as 24.7% at 1.0 mg/mL. Four SSE fractions were isolated by Sephadex G-25 gel chromatography; the F2 fraction showed the highest DPPH radical scavenging activity. The F2 fraction was separated by reverse-phase high performance liquid chromatography (HPLC) using an octadecylsilane (ODS) column, yielding a purified antioxidant substance with a DPPH radical scavenging activity of 64.41% at 1.0 mg/mL, representing a 2.64-fold increase in the scavenging activity of SSE purified by the 3-step procedure. The amino acid compositions showed that purified SSE was rich in taurine, glycine, glutamic acid and alanine. The purified SSE significantly elevated 2',7'-dichlorodihydrofluororescein diacetate (DCFH-DA) fluorescence probe, which confirms its effective radical scavenging potential in cellular ROS. In addition, the SSE significantly inhibited oxidative damage of purified genomic DNA. These results suggest that a purified antioxidant substance from SSE can be used as a potential natural compound-based antioxidant in the functional food and pharmaceutical industries.