The absolute stereochemistry of FD-594 1, a new cytotoxic antibiotic, was determined by X-ray diffraction, and its conformation was studied by CD and NMR spectroscopy. The aglycon part of 1 was found to have (3R,6S,7S) configuration. Particularly interesting was the solvent-dependent atropisomerism of 1 and related compounds. The CD spectra of 1 exhibited in two solvent systems almost opposite mirror-image curves depending on the solvent. While a large negative Cotton effect (Deltaepsilon = -33.9, 279 nm) was observed in CHCl(3), a similar positive Cotton effect (Deltaepsilon = 38.9, 279 nm) appeared in methanol most probably due to dramatic conformational changes. Similar chiroptical reversal was observed in aglycon 2 and aglycon methyl ether 4. These results can be best described in terms of solvent-dependent atropisomerism. This constitutes the first observation of solvent-dependent atropisomerism of a natural product. The crucial factor that perturbs the stable conformation in different solvents is discussed on the basis of molecular mechanics calculations.