In previous work, a series of daphnane-type diterpenes isolated from D. genkwa were found to exhibit marked cytotoxicity against a variety of cancer cell lines.(12) In further related work on this genus, an EtOH extract of D. giraldii was investigated by means of solvent extraction and conventional chromatography, followed by chiral HPLC analysis. As a result, compounds 1 and 2 were found to afford four pairs of enantiomers (1a-1/1a-2, 1b-1/1b-2, 2a-1/2a-2, and 2b-1/2b-2). In addition, two pairs of epimers (3a/3b and 4a/4b) were also obtained. The absolute configurations of compounds 1–3 were determined by producing [Rh2(OCOCF3)4] complexes and by the comparison of ECD and optical rotation data to the calculated values. In this report, the isolation, structural elucidation, and cytotoxic activity evaluation of these compounds against four human tumor cell lines are described.