Methanol extracts of the corms of Hypoxis rooperi and H. latifolia were studied for their hypoxoside content by an in-line sorption enrichment HPLC technique [Kruger et al., J. Chromatogr., 612 (1993) 191]. Hypoxoside is the trivial name for (E)-1,5-bis(3'-hydroxy-4'-O-beta-D-glucopyranosyl-phenyl) pent-1-en-4-yne and rooperol the aglucone obtained from beta-glucosidase treatment. Hypoxoside and rooperol analogues containing 4, 3 and 2 hydroxyl groups resolved as separate peaks with the proportion of the latter two markedly higher in H. latifolia than in H. rooperi. After oral ingestion of hypoxoside by humans, no hypoxoside or rooperol appeared in the serum. Only rooperol was present in the faeces. The serum and urine contained at least three phase II metabolite peaks. Selective enzyme hydrolysis showed that they represent the diglucuronide, disulfate and glucuronide-sulfate conjugates of all three rooperol analogues.