The metabolism of certain compounds by rat hepatic microsomal preparations obtained from acetone-treated rats is greater than that measured in microsomes obtained from control rats. Since the lipid composition of membranes can significantly influence metabolism catalyzed by membrane-bound enzymes, studies were conducted in order to determine whether acetone-induced alterations in microsomal metabolism are associated with changes in the lipid composition of these membranes. Phospholipid and cholesterol concentrations were measured in extracts of rat liver microsomes obtained from rats treated with acetone, sodium phenobarbital, or vehicle. Acetone treatment did not alter the phospholipid and cholesterol content of the microsomes, whereas sodium phenobarbital (positive control) caused a significant decrease in cholesterol content/mg microsomal protein. These data indicate that acetone-induced alterations in microsomal metabolism may not be attributed to changes in the phospholipid and cholesterol content of the microsomes.