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Abstract: Seven steroidal glycosides, along with one known glycoside, were isolated from the rhizomes of Ruscus hypophyllum (Liliaceae). Comprehensive spectroscopic analysis, including 2D NMR spectroscopy, and the results of acid hydrolysis allowed the chemical structures of the compounds to be assigned as (23S,25R)-23-hydroxyspirost-5-en-3β-yl O-α-l-rhamnopyranosyl-(1→4)-β-d-glucopyranoside (1), 1β-hydroxyspirosta-5,25(27)-dien-3β-yl O-α-l-rhamnopyranosyl-(1→4)-β-d-glucopyranoside (2), (22S)-16β,22-dihydroxycholest-5-en-3β-yl O-α-l-rhamnopyranosyl-(1→4)-β-d-glucopyranoside (3), (22S)-16β-[(β-d-glucopyranosyl)oxy]-22-hydroxycholest-5-en-3β-yl O-α-l-rhamnopyranosyl-(1→4)-β-d-glucopyranoside (4), (22S)-16β-[(β-d-glucopyranosyl)oxy]-22-hydroxycholest-5-en-3β-yl β-d-glucopyranoside (5), (22S)-16β-[(β-d-glucopyranosyl)oxy]-3β,22-dihydroxycholest-5-en-1β-yl O-α-l-rhamnopyranosyl-(1→2)-(3,4-di-O-acetyl-β-d-xylopyranoside) (6), and (22S)-16β-[(β-d-glucopyranosyl)oxy]-3β,22-dihydroxycholest-5-en-1β-yl O-α-l-rhamnopyranosyl-(1→2)-O-[β-d-xylopyranosyl-(1→3)]-β-d-xylopyranoside (7), respectively. This is the first isolation of a series of cholestane glycosides from a Ruscus species. [Copyright &y& Elsevier]