The distribution, labeling and interrelationship of microsomal and lysosomal dolichol and dolichyl-P in rat liver was investigated. After membrane induction with phenobarbital, N-nitrosodiethylamine and diethylhexylphthalate, the amount of microsomal and lysosomal dolichols are modulated independently. Liposomal labeled dolichol injected into the portal vein appears only in lysosomes and even after 8 days is still limited to the lysosomes. After in vivo labeling with [3H]mevalonate, high initial labeling of dolichol and dolichyl-P is present in microsomes and the labeling in microsomes is greater than that in lysosomes even after 8 h. The results demonstrate compartmentalization of the intracellular dolichols in hepatocytes. These lipids may have independent roles at different membrane locations.