The current study sought to determine the effects of Colpomenia sinuosa and Sargassum linifolium aqueous extracts on non-enzymatic and enzymatic antioxidants in Spinacia oleracea L. leaf extract. Glutathione synthase (GSS, EC: 6.3.2.3) and -glutamyl cysteine synthetase (-GCS, EC: 6.3.2.2) are involved in the glutathione biosynthesis process. Treatment of S. oleracea with seaweed extracts increased the level of reduced glutathione (GSH), oxidized glutathione (GSSG), and total glutathione at lower concentrations. Total phenols and total flavonoids in S. oleracea leaves accumulated more rapidly after treatment with seaweed extracts. The activity of the enzyme's phenylalanine ammonia lyase (PAL, EC: 4.3.1.25), chalcone synthase (CHS, EC: 2.3. 1.74), and chalcone isomerase (CHI, EC: 5.5.1.6) involved in the production of phenylpropanoid and flavonoid in S. oleracea leaves rose in a dose-dependent manner. Glutathione reductase (GR, EC 1.6.4.2), glutathione peroxidase (GPX, EC: 1.11.1.9), and glutathione-S-transferase (GST, EC: 2.5.1.18) are examples of antioxidant enzymes whose activity were also elevated by the treatment. In comparison to untreated plants, the treated plants' S. oleracea leaf extract significantly reduced superoxide anion, 1, 1-diphenyl-2-picrylhydrazyl (DPPH), and hydroxyl radicals. In light of this, the current findings imply that the use of seaweed extracts was found to boost the activity of the antioxidant system in Spinacea oleracea