AIM　To study the metabolic pathway of chiral clausenamide in the rat and understand its stereoselectivity. METHODS　The urine, feces and blood of rat were gathered after the drug was administered, the known metabolites were analyzed by HPLC-DAD and one unknown metabolite was elucidated by using LC-MS analysis. Metabolic stereoselectivity was determined by comparing the metabolic results of (+)- and (-)-clausenamide. RESULTS　Six known metabolites were determined and one unknown metabolite was elucidated as N-demethylclausenamide. The metabolic stereoselectivity was shown distinctly. CONCLUSION　Chiral clausenamide was mainly metabolized by hydroxylation in liver and the biotransformation exhibited pronounced substrate stereoselectivity.