The chemical composition of Populus balsamifera essential oils obtained from spring buds, fall buds, and young leaves were determined by GC and GC-MS analyses. The major constituent, (+)-alpha-bisabolol, a rare sesquiterpene, was isolated from spring oil using reverse-phase preparative HPLC. The cytotoxic activity of balsam poplar oils and isolated (+)-alpha-bisabolol was assessed in vitro against human lung carcinoma (A549) and colorectal adenocarcinoma (DLD-1) cell lines. Essential oils were cytotoxic with IC50 ranging from 35 to 50 microg/mL. (+)-alpha-Bisabolol exhibited pronounced activity (IC50 14 microg/mL) against both cancer cell lines. It also exhibited interesting cytotoxic activity (IC50 23 microg/mL) against human glioma (U251), higher than the one observed for (-)-alpha-bisabolol (IC50 34 microg/mL), which is known for its apoptosis-inducing effect against glioma cells.