ABSTRACTSpinacia oleracea is a green leafy vegetable widely consumed for its nutritional value. The present research work is focused on spinacia oleracea seed extract for evaluation of in vitro antioxidant and in vivo anxiolytic activities. Spinach leaves antioxidant properties are well documented but the bioactive potential of seeds remains unexplored. Hence this research work aims to explore the antioxidant and anxiolytic effects of Spinacia oleracea seed extract using standard in vitro and in vivo methods. The in vitro antioxidant potential of the spinacia oleracea seed extract at dose of 50,100,300 and 500µg/ml were assayed using hydroxyl radical scavenging, nitric oxide, and DPPH radical scavenging assay and results are compared with standard reference drug Gallic acid at concentration 2.5µg/ml. In behavioral studies, spinacia oleracea seed extract at dose of 75mg/kg and 150mg/kg was tested on albino mice subjected to standard behavioral paradigms such as elevated plus maze, open field test, hole board test, and y-maze. The results of these effects were compared to standard anxiolytic drug diazepam. The results obtained were encouraging and indicated significant antioxidant and anxiolytic effect. The FTIR studies revealed that the seed extract was rich in phenolic compounds known for its antioxidant property.Keywords: Spinacia oleracea; antioxidant; DPPH; anxiolytic; elevated plus maze; FTIR