Artesunate is a potent and rapidly acting blood schizontocide used to treat chloroquine resistant malaria. Artesunate has been reported to cause embryo, reduced reproductive capacity, hepatotoxicity, neurotoxicity and hematological abnormalities. Previously toxicity studies on artesunate have been done in 2–10 mg/kg dose range mostly for 7 days, scientific studies on sub-chronic exposure of artesunate is not been reported so for. The present study evaluates sub-chronic safety profile of artesunate on 45 days oral administration at 2, 4 and 8 mg/kg/day. Authentication of artesunate has been done by color test, pH, melting point, loss on drying, UVmax, TLC and HPLC study. Artesunate has non-significant effect on liver and kidney weight. Serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), serum alkaline phosphate (ALP), cholesterol (TC), triglyceride (TG), total protein, albumin, bilirubin, creatinine, urea and glucose content were estimated after 45 days treatment along with hematological screening. Artesunate treatment for 45 days significantly increased (p < 0.05–0.001) SGOT, SGPT, ALP, TC, TG, total bilirubin, glucose level at 8 mg/kg/day dose. It has non-significant effect on serum total protein, albumin, creatinine and urea. Hemoglobin, total RBC, platelet, lymphocytes, basophil, mean cell volume and mean corpuscular hemoglobin concentration have not changed but total WBC, neutrophil, eosinophil, packed cell volume and mean cell hemoglobin were increased significantly (p < 0.01) at 8 mg/kg/day dose. Artesunate treatment at 4 and 8 mg/kg/day showed sinusoidal dilation, cytoplasmic vaculation, focal necrosis, sinusoidal congestion and extensive inflammatory changes, whereas kidney was free of any deleterious effect.ConclusionSub-chronic exposure of artesunate at 8 mg/kg/day dose for 45 days period cause hepatic damage along with hematological abnormalities signifying safety concern.