Two novel furostanol saponins were isolated from the fresh tubers of Ophiopogon japonicus. Comprehensive spectroscopic analysis allowed the chemical structures of the compounds to be assigned as (25R)-26-[(O-beta-d-glucopyranosyl-(1 --> 2)-beta-d-glucopyranosyl)]-22alpha-hydroxyfurost-5-ene-3-O-beta-d-xylopyranosyl-(1 --> 4)-O-[alpha-l-rhamnopyranosyl-(1 --> 2)]-beta-d-glucopyranoside (1, ophiopogonin F) and (25R)-26-[(O-beta-d-glucopyranosyl-(1 --> 6)-beta-d-glucopyranosyl)]-22alpha-hydroxyfurost-5-ene-3-O-beta-d-xylopyranosyl-(1 --> 4)-O-[alpha-l-rhamnopyranosyl-(1 --> 2)]-beta-d-glucopyranoside (2, ophiopogonin G). The rare furostanol saponins with two glucosyl residues at C-26 position were isolated from the natural source for the first time.