Further study of constituents from the aerial parts of Asclepias tuberosa afforded twenty-two new steroidal glycosides along with tuberoside B5 and G5. These glycosides were confirmed to contain 8,12;8,20-diepoxy-8,14-secopregnanes, tuberogenin and its congeners, as their aglycones. The structure of each of these compounds was elucidated based on the interpretation of NMR and MS measurements and from chemical evidence.