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Phenobarbital (PB) was administered to a positive control group by oral gavage at a daily dose of 80\u2009mg\u2009kg<jats:sup>\u22121<\/jats:sup>. TBA caused, on day 14, a reduction in circulating T3 (12\u201315% decrease) and a dose\u2010dependent reduction in T4 (13\u201322% decrease), with no evidence of thyroid pathology. Two of five livers examined in the 20\u2009mg\u2009TBA\u2009ml<jats:sup>\u22121<\/jats:sup> dose group showed mild, diffuse centrilobular hypertrophy. On day 14, Cyp 7\u2010benzoxyresorufin\u2010<jats:italic>O<\/jats:italic>\u2010debenzylase activity was significantly induced 12\u2010fold by TBA at 20\u2009mg\u2009ml<jats:sup>\u22121<\/jats:sup>, and 1.8\u2010fold at the 2.0\u2009mg\u2009TBA\u2009ml<jats:sup>\u22121<\/jats:sup> concentration. Cyp 7\u2010pentoxyresorufin\u2010<jats:italic>O<\/jats:italic>\u2010dealkylase activity was slightly induced (2.1\u2010fold) by 20\u2009mg\u2009TBA\u2009ml<jats:sup>\u22121<\/jats:sup> on day 14. Quantitative PCR analysis of gene transcripts showed a significant induction of Cyp2b10 and ST1a1 with both TBA concentrations, and a slight induction of Cyp2b9 at 20\u2009mg\u2009TBA\u2009ml<jats:sup>\u22121<\/jats:sup> only. PB induced all phase I and phase II gene transcripts except for Cyp1a1 and Cyp2b9. These findings suggest that TBA, at and below doses used in chronic studies, is an inducer of phase I and phase II liver enzymes, with resulting decreases in circulating thyroid hormones in B6C3F1 mice. 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