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Background: Medicinal plants have been considered a suitable source for finding new pharmaceutical chemicals, and identification of chemical composition is the first step towards uncovering the nature of bioactive compounds. Objectives: Scrophularia striata is used to treat various infections in traditional medicine. The aim of this study was to evaluate phytochemical and biological analysis of S. striata collected from Ilam. Methods: In the present study, the hydro distilled volatile oils of aerial parts of S. striata were investigated by GC-MS and GC-FID. The total phenolic and total flavonoid content of hydro-alcoholic extract of S. striata were measured by the Folin-Ciocalteu and aluminium chloride colorimetric method, respectively. The antioxidant activity of hydro-alcoholic extract was studied by using the 2,2-diphenyl-1-picrylhydrazyl assay. Results: In total, 26 compounds were identified and the main identified compounds of the essential oil were n-hexane (16.3%), caryophyllene oxide (15.36%), spathulenol (13.1%), α-cadinol (12.35%), and docosane (6.33%). The hydro-alcoholic extract showed moderate antioxidant activity with IC50 = 85 µg mL -1 . The total phenolic content of extract was 15.9 mg gallic acid equivalents/g dry weight, and the total flavonoid content of extract was 1.8 mg quercetin/g dry weight.Conclusions: The data of this study suggests that the essential oil and hydro-alcoholic extract of S. striata could be considered a natural antioxidant agent to preserve food and treat infections in food and pharmaceutical industry. In addition, the data of this study can be used for further studies on the taxonomy of Scrophularia genus.