A new monodesmosidic spirostanol saponin, along with three known saponins was isolated from Agave macroacantha Zucc leaves. The structure of the new saponin was established as hecogenin-3-O-α-l-rhamnopyranosyl (1→4) β-d-xylopyranosyl (1→3)[β-d-glucopyranosyl (1→2)] β-d-glucopyranosyl (1→4) β-d-galactopyranoside. The 1 H and 13 C resonances of the four compounds were assigned using a combination of 1D and 2D NMR techniques including 1 H, 13 C, COSY, TOCSY, ROESY, HSQC and HMBC NMR and confirmed by mass spectrometry.