Enzyme distribution in rat liver lobes was investigated using liver homogenate as the enzyme source. When the content or the activity was expressed on the basis of unit (gram) wet weight of liver tissue, the protein concentration was almost the same throughout the liver, but several enzymes were distributed heterogeneously within the liver. Cytochrome P-450 monooxygenase activity was higher in the median and right lobes of livers, while the left lobe contained a higher concentration of mitochondrial enzymes. Phenobarbital induced cytochrome P-450 monooxygenase, and the activity was still higher in the median and right lobes even after the pretreatment of rats with phenobarbital. On the other hand, administration of beta-naphthoflavone resulted in a uniform increase of cytochrome P-450 (P-448) content throughout the liver to almost the same concentration. Increase of cytochrome P-448-dependent O-dealkylation activity of 7-alkoxycoumarin by the administration of beta-naphthoflavone was much greater in the left lobe compared to that in the median and the right lobes. Heterogeneous distribution of the enzymes in various liver lobes was also determined employing liver samples obtained from various physiological states of rats (fasting, glucose refeeding after fasting and hyperthyroidism), although the cytochrome P-450 content and drug-metabolizing activity were altered markedly depending upon the physiological states of the rats.