To examine if chronic oral administration of phenobarbital (PB), a widely used anti-epileptic drug, has any genotoxic and cytotoxic potential in mice, a mammalian model, cytogenetic assays through several endpoints such as chromosome aberrations, induction of micronuclei, mitotic index of bone marrow cells, sperm-head anomaly in testis and enzymatic assays of several toxicity marker enzymes have been conducted by use of standard techniques. Mice of both treated (chronically receiving an oral dose of PB at 1.2 mg/kg bw) and control (without receiving PB) groups were sacrificed at 7, 15, 30, 60, 90 and 120 days for the study with all the above-mentioned protocols. Further, total protein profiles in liver of both control and treated mice were analyzed through the SDS-PAGE technique at day 60. The results of all these studies, when compared with controls, showed that PB has both genotoxic and cytotoxic potential in apparently increasing intensity at longer periods of chronic feeding in mice, which would warrant due consideration in its long-term use on human subjects.