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A steroidal saponin isolated from the tuber of Ophiopogon japonicus. The structure of the saponin was elucidated by several one-and two-dimensional NMR experiments to beThe isolation and structure elucidation of several steroidal saponins of Ophiopogonis tuber is well Ruscogenin and diosgenin saponins have been determined as the major glycosides, whereas the minor glycosides are their (25s) isomers. Constituents of Ophiopogon japonicus tuber are ruscogenin and diosgenin glycosides, at which, in the case of ruscogenin, the sugar moiety is linked monodesmosidically to the hydroxyl group at C-l. We describe the isolation and structure elucidation of a new bisdesmosidic (25S)-ruscogenin saponin from the tuber of Ophiopogon japonicus, which has sugar moieties linked to the hydroxyl groups at C-1 and C-3. This saponin has been described before in the subterranean part of Liriope pkztyphyllu Wang et Tang, one congener crude drug of Ophiopogonis tuber [']. In this paper the full assignment of the proton-and carbon-NMR signals is given.