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Objective: To assess the toxic potential of Dissotis rotundifolia (D. rotundifolia) whole plant extract in Spraque–Dawley rats within a 2-week period of administration. Methods: Methanolic extract of D. rotundifolia was administered orally once daily at dose levels of 0, 100, 300 and 1000 mg/kg body weight for 14 days. Toxicity was assessed using mortality, clinical signs, body and organ weights, hematological indices, serum chemistry parameters and histopathological analyses. Results: There were no treatment-related mortalities or differences in clinical signs, hematology and serum biochemistry. This was confirmed by micrographs obtained from histopathological analysis. Conclusions: The results obtained from the sub-acute toxicological assessment of D. rotundifolia extract suggest that the extract is non-toxic at doses up to 1000 mg/kg/day administered for a period of 14 days.