The metabolism of lidocaine was studied using microsomes from extrahepatic tissues of rats, including lung, kidney and brain, or using a reconstituted system with purified CYP2B1 and CYP4B. Rat pulmonary microsomes metabolized lidocaine to an N-deethylated metabolite, monoethylglycinexylidide (MEGX). Renal microsomes produced MEGX and 3-hydroxylidocaine (3-OH LID), although the rate of MEGX formation was much lower in renal than in pulmonary microsomes. Other metabolites were not detected. Lidocaine was not metabolized by brain microsomes. In extrahepatic tissues, pulmonary microsomes had the highest activity. Hence, two major forms of cytochrome P450 isozymes, CYP2B1 and CYP4B1, in rat pulmonary microsomes were used for further study. The study with a reconstituted system using purified cytochrome P450 isozymes revealed that only CYP2B1 showed lidocaine deethylation activity; the other form of cytochrome P450 in the lung, CYP4B1, did not. The Michaelis-Menten constant for lidocaine N-deethylation by rat pulmonary microsomes was 0.27 mM. Antibody against CYP2B1 completely inhibited the formation of MEGX by pulmonary microsomes. These results suggest that lidocaine is metabolized by rat lung, including CYP2B1.