Tagetes erecta Linn. has been used in folk medicine for the cure of eye diseases, colds conjunctivitis, rheumatism, coughs, bleeding piles and ulcers. In addition, in our previous study, we have reported the potent antimicrobial, cytotoxic and insecticidal activities of chloroform fraction of Tagetes erecta flower. However, its other pharmacological effects have not yet been elucidated clearly. The aim of this study was to investigate the subacute toxicity of chloroform fraction prepared from ethanol extract of Tagetes erecta flower by solvent-solvent partitioning method. The subacute toxicity of chloroform fraction was evaluated on Long Evan?s rats at 200 and 400 mg/kg doses and the results obtained from chloroform fraction treated rats were compared with untreated controls. Treatment of chloroform fraction at 200 and 400 mg/kg doses, did not make any significant alterations on the hematological and biochemical parameters of rats when data were compared with that of untreated controls. Histopathological examination also showed no detectable changes in liver, kidney, heart and lung of chloroform fraction treated rats. This study revealed that the chloroform fraction of Tagetes erecta had no toxic effects. The results validate the traditional use of this plant in indigenous system of medicine.