Diplotaenia turcica is widely utilized in conventional treatment in the east of Turkey. Due to the insufficient data on the safety profile, the acute and sub acute toxicity of Diplotaenia turcica was determined. Furthermore, total phenolic content, flavonoid content and antioxidant activity of the extract were determined. The acute toxicity of hydroalcoholic root extract of Diplotaenia turcica (HREDT) was evaluated in mice after administration of single oral dose at the 5000 mg/kg (acute model) and rat after 28 days orally administration at the dose range of 250, 500 and 1000 mg/kg (subacute model). According to result, the LD50 value of HREDT was found to be greater than 5000 mg/kg. In sub acute toxicity study, no statistically significant differences were observed in the values of hematological and pathological parameters in comparison with control group. It was determined that the glucose, triglyceride, cholesterol and LDL levels exerted a significant effect depending on the HREDT doses. The results from the present study found out that HREDT did not produce any toxic effects or deaths in animals for both single and chronic administration. Additionally, HREDT showed moderate antioxidant activities and exhibited relatively notable total phenolic content.