Objective: To evaluate the safety of ethanolic seed extract of Eugenia jambolana (EJSE) using acute and sub-chronic toxicity assays in Swiss albino mice as per Organisation for Economic Co-operation and Development (OECD) guidelines. Methods: Possible behavioral changes and lethality were observed in mice administered a single dose [1 000, 2 000, 3 000, 4 000 or 5 000mg/kg body weight (BW)] of EJSE. Plasma levels of metabolic, hepatic, cardiac and renal function markers, electrolytes, blood count and histopathology of major organs were monitored in mice chronically treated with EJSE (1 000, 2 000 or 3 000 mg/kg BW) for 28 days. Results: Since no mortality was recorded in the acute toxicity evaluation up to a dose of 5 000 mg/kg bodyweight of EJSE, 50%lethal dose (LD50) was assumed to be >5 000 mg/kg BW. In the sub-chronic toxicity evaluation, no adverse observations were recorded in mice administered with 2 000 mg/kg BW of EJSE;however at 3 000 mg/kg BW dose, moderately significant increase in the plasma levels of urea and creatinine was observed. Hence, the lowest observable adverse effect level (LOAEL) for EJSE was found to be 3 000 mg/kg BW and the no observable adverse effect level (NOAEL) was adjudged as 2 000 mg/kg BW. Conclusions: It can be concluded from this study that, orally administered EJSE is safe up to a10 fold higher dose than its reported therapeutic dose.