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A new triterpenoid saponin, 3β-O-{β-D-glucopyranosyl-( 1 → 4)-[β-D-glucopyranosyl-(1 → 2)]-α-L-arabinopyranosyl}-16α, 28-dihydroxyolean-12-en-30-oic acid 30-O-β-D-glucopyranosyl ester (ardisicrenoside N. 1), together with two known saponins, ardisicrenoside C (2) and D (3), were isolated from the roots of Ardisia crenata Sim. Their structures were elucidated by extensive spectral analysis and chemical evidences. Saponins 1 showed cytotoxicity against MCI-7 and NCI-H460 cancer cell lines at 11.0 μmol/L and 22.1 μmol/L in vitro.