The present study was carried out to evaluate the antimitotic potential of Coffea arabica seeds by standard assay method using Allium cepa root meristem model. The result of the study showed that the Coffea arabica seeds extract had excellent anti-mitotic activity that was comparable to the standard methotrexate. In Allium assay, aqueous extract of seeds of Coffea arabica and methotrexate showed significant concentration dependent inhibitory influence against the dividing cells of Allium roots and decreased root growth, length and mitotic index as compared to control distilled water. Maximum numbers of non-dividing cells were observed in higher dose. As a result of this cells arrest in mitosis and eventually die by apoptosis. There was a marked decrease in the mitotic index. Thus, it can assume the possible mechanism of the anticancer activity of Coffea arabica seeds extract may be due to the presence of phenolic compounds in the extract may contribute in cell growth inhibition. Our findings support the reported therapeutic use of Coffea arabica seeds as an antimitotic agent in the Indian system of medicine.