Study was made of brassinosteroids and phytosterols in unsaponifiable lipid obtained from the seeds of Cannabis sativa L. Bioactive substances in a rice-lamina inclination test were highly purified, derivatized and analyzed by GC-MS. The brassinosteroid, teasterone, was identified, suggesting for the first time its possible presence as a fatty acid ester in dicot plants. Phytosterols in the seeds were found to contain six phytosterols, cholesterol, campesterol, campestanol, stigmasterol, sitosterol and sitostanol, with sitosterol and campesterol present in the largest amounts. Campestanol and sitostanol were identified in the seeds for the first time. The structural relationship between brassinosteroids and phytosterols in the seeds is discussed from a biosynthetic point of view.