Steroidal constituents from the roots and stems of Asclepias fruticosa L. were investigated separately. From the roots, twelve pregnane pentaosides and uzarigenin β-sophoroside were isolated together with three known coroglaucigenin and corotoxigenin glycosides. Pregnane glycosides were composed of ikemagenin or kidjolanin as an aglycone, and D-digitoxose, D-cymarose, D-oleandrose and terminal D-glucose as component sugars. Among the constitutents from the stems, cardenolides show a similar pattern to those from the leaves. 17α-Hydroxycalactin and 17α-hydroxyafroside were newly obtained along with known doubly linked and normally linked glycosides. Two pregnane glycosides and uzarigenin β-sophoroside obtained from the roots were also isolated.