Aim: To evaluate the sub-acute toxicity profile of fractions of Vitex doniana using adult albino rats. Study Design: An experimental study that lasted for 28 days. Place and Duration of Study: Department of Medical Laboratory Sciences and Animal House Unit, College of Medicine, University of Nigeria Enugu Campus, between January 2019 and March 2019. Methodology: Methanol, ethyl acetate and hexane fractions of V. doniana were orally administered to 6 groups of animals at doses of 150 mg/kg and 300 mg/kg body weight for each fraction, for 28 days. At the end of the administration, the animals were sacrificed. Body weight changes, Relative Organ Weights (ROW), haematological analysis, serum biochemical assay and histopathological changes of vital organs were used to evaluate the potential toxic effects of the fractions after treatments. Results: The fractions had no adverse effects on the body weights, ROW and gross appearance of the tissues. There were no significant changes in the haematological and biochemical parameters of the treated animals compared to the controls. Histological assessment of the excised organs also showed no overt pathological changes apart from thickened alveolar septa in the lungs of the animals that received methanol and ethyl acetate fractions. Conclusion: The polar fractions of V. doniana at the doses tested did not induce any toxic changes in the treated animals.