Lagenaria siceraria (Family: Cucurbitaceae), sometimes referred to as bottle gourd, calabash, doodhi, and lauki, is a plant that is widely utilised as a fruit and vegetable throughout India. The objective of the current study was to assess the toxicity, in vitro antioxidant activity, and Phytochemical content of Lagenaria siceraria seeds. The results indicate that the extract lacks polysaccharides, proteins, steroids, tannins, and cardiac glycosides, while it does contain cardiac glycosides, reducing sugar, flavonoids, and carbohydrates. The total flavonoid concentration of the extract, which was measured at 38.64 mg/g, exhibited a robust level of antioxidant activity. The concentration-dependent scavenging of DPPH radicals by the extract was observed, with the highest efficacy being exhibited by the ether+ alcoholic extract. As per the outcomes of the toxicity evaluation, the extract exhibited no toxicity; mice did not manifest any fatality or unfavourable impacts at quantities as high as 5000 mg/kg. It can be inferred that the LD50 value of the extract is greater than 5000mg/kg. The findings suggest that the extract derived from Lagenaria siceraria seeds exhibits considerable potential as an antioxidant agent and possesses desirable safety characteristics. Therefore, further investigation into its pharmacological and toxicological properties is warranted.