The present study aimed to evaluate the acute and subacute toxicity profiles of Euphorbia hirta leaves extract in albino rats using the methods described in the guidelines of the OECD. The acute toxicity was performed where the limit dose of 2000 mg/kg body weight used. Observations were made and recorded for 24 h, and once daily further for a period of 14 days. The rats were weighed and various observations, like mortality, behavior, injury, or any signs of illness were conducted once daily during the period. For subacute study, four groups of 6 animals (Male rats) received distilled water (control), and 100, 200 and 400 mg/kg of freshly-prepared extracts, respectively, every 24 h orally for 28 days. At the end of each study, hematological analysis and biochemical parameters were evaluated. Histopathological examination of vital organs (Liver and kidney) of the animals were taken for gross findings, compared to controls. There was no significant difference (p > 0.05) observed in the relative organs, body weights, hematological, biochemical parameters, and gross abnormalities, compared to the control. No mortality was recorded. Therefore, analysis of results may lead to the conclusion that the medium-term oral administration of the Euphorbia hirta leaves extract (EHLE) for 28 days does not cause toxicity. On the basis of acute and sub-acute toxicity studies, the minimal effective dose of 200mg/kg is taken for efficacy (anti-cancer) studies.