Abstract Regeneration of adventitious shoots from young leaves of Datura metel is described. Shoot buds developed on MS medium with 2 mg/l and elongated on hormone-free solid basal medium. The microshoots failed to produce alkaloids, but a number of steroidal compounds were detected. The C 28 sterol 3 β,24 ξ-dihydroxy-ergosta-5,25-dienolide, regarded as the precursor of withanolides and related steroidal lactones, was identified. The withanolide 12-deoxywithastramonolide was also detected.