Approx. one-third of an i.v. dose of 14C-benzo[a]pyrene was excreted within four hours in the bile of guinea-pigs fed a normal diet. The extent of excretion was not altered by feeding high-fat or high-cholesterol diets. Hepatic cytochromes P-450 and b5, and benzo[a]pyrene hydroxylase activity were unaltered by the administration of high-fat and high-cholesterol diets. Pretreatment with low oral doses of benzo[a]pyrene (6 X 3 mg/kg) did not induce these parameters in animals given any of the diets. High-fat and high-cholesterol diets altered the pattern of benzo[a]pyrene metabolites in the bile, with significantly increased excretion of dihydrodiol glucuronides in both the high-fat and high-cholesterol groups. Hepatic epoxide hydrolase activity and glutathione content were unaltered by the high-fat or high-cholesterol diets, and therefore cannot explain the alteration in the profile of biliary metabolites of benzo[a]pyrene. The altered pattern of biliary excretion in animals fed high-fat or high-cholesterol diets would lead to an increase in the delivery to the colon of dihydrodiol metabolites of benzo[a]pyrene.