Chemical investigation of ecdysteroidal constituents of the roots and stems of Cyathula officinalis led to the isolation of two cyasterone stereoisomers, 2 and 3, together with the known cyasterone 1. The structures of compounds 2 and 3 were determined to be 28-epi-cyasterone and 25-epi-28-epi-cyasterone, respectively, by means of spectroscopic analysis. X-Ray structures of 1 and 2 confirmed the 24S,25S,28R configuration for 1 and 24S,25S,28S for 2.