Due to the emergence of multi-drug resistant Mycobacterium tuberculosis strains, there is an urgent need to search for new antimycobacterial drugs. The present work describes the bioactivity-guided fractionation of Chamaedora tepejilote hexane extract which led to the isolation and the characterization of squalene, farnesol, methylic ester of hexadecanoic acid, beta-sitosterol and ursolic acid. Activity against Mycobacterium tuberculosis H37Rv, determined by radiorespirometric Bactec assay, showed ursolic acid, squalene and farnesol to produce a M. tuberculosis growth inhibition of 99% at a concentration of 100 mu g/mL. Copyright (c) 2005 John Wiley & Sons, Ltd.