Steroidal compounds in the roots of Asclepias tuberosa were investigated and 17alpha-hydroxyandrosta-4,6,15-trien-3-one 17-O-alpha-L-arabinopyranosyl-(1-->6)-beta-D-glucopyranoside, termed ascandroside, was isolated from the CHCl3-soluble fraction. Among five doubly-linked cardenolide glycosides, two were identified as 3'-spiro-linked thiazolidinone (4) and S-oxythiazolidinone derivatives (5) of delta5-calotropin. The stereochemistry at the C-3' in these two cardenolides is discussed. 3'-O-beta-D-Glucopyranosyl-delta5-calotropin (3) was also isolated along with A5-calotropin and its 3'-acetate. Nine glycosides of uzarigenin, coroglaucigenin and corotoxigenin were identified.