To utilize fish waste, black pomfret, Parastromateus niger viscera was analysed for its proximate and amino acid composition followed by hydrolysis using various proteases to extract antioxidant peptide. Antioxidant activities of the crude hydrolysate was evaluated using DPPH (54%), metal chelating (78.6%) at a concentration of 1 mg/mL, whereas the reducing power assay was done with different concentration (0.5-2.5 mg/mL) and the activity also increased with increasing concentration (0.021-0.068). Furthermore, the hydrolysate was purified by diethylaminoethyl (DEAE) ion-exchange and Sephadex G-25 gel filtration chromatography. Finally, the purified peptide had a mass of 701.9 Da, and the amino acid sequence was identified as Ala-Met-Thr-Gly-Leu-Glu-Ala using electrospray ionization-tandem mass spectrometry (ESI-MS/MS). Moreover, the protection ability of the peptide toward hydroxyl radical-induced oxidative DNA damage and inhibiting lipid peroxidation was evaluated and compared with natural antioxidant ${alpha}$-tocopherol.