Abstract: To study the chemical constituents from traditional Chinese herb Ophiopogon japonicus (Thunb.) Ker-Gawl., a new steroidal glycoside, named ophiopojaponin C (1), together with two known ones, was isolated by column chromatography. Spectroscopic and chemical evidence revealed the structures to be ophiopogenin 3-O-[α-L-rhamnopyranosyl(1→2)]-β-D-xylopyranosyl　(1→4)-β-D-glucopyranoside (1), diosgenin 3-O-[2-O-acetyl-α-L-rhamnopyranosyl　(1→2)]-β-D-xylopyranosyl　(1→3)-β-D-glucopyranoside (2), and ruscogenin 1-O-　[2-O-acetyl-α-L-rhamnopyranosyl(1→2)]-β-D-　xylopyranosyl(1→3)-β-D-fucopyranoside (3).

Key words: C27 steroidal glycosides, Liliaceae, Ophiopogon japonicus.