A new lanostane-type triterpene glycoside, impatienside A (1), was isolated from the sea cucumber Holothuria impatiens, together with a structurally related, known compound, bivittoside D (2). Their structures were elucidated by in-depth spectroscopic and mass-spectrometric methods, including (1)H-, (13)C-, and 2D-NMR, ESI-MS, and HR-ESI-MS experiments, as well as by chemical evidence. Compounds 1 and 2 possess the same hexasaccharide moieties, but differ slightly in their holostane-type triterpene aglycone. The two glycosides were found to exhibit in vitro cytotoxicities similar to or better than those of the potent anticancer drug etoposide (V-16) against seven different human tumor cells, with IC50 values of 0.37-2.75 microg/ml.