The bark extract of Acacia mearnsii showed strong lipase and α-amylase inhibition activities. Fractionation of the extract by column chromatography and subsequent (13)C NMR and MALDI-TOF-MS analysis revealed that the active substances are proanthocyanidin oligomers mainly composed of 5-deoxyflavan-3-ol units. In addition, 4'-O-methylrobinetinidol 3'-O-β-D-glucopyranoside, fisetinidol-(4α,6)-gallocatechin, and epirobinetinidol-(4β,8)-catechin were isolated as new compounds, and their structures were determined from spectroscopic data. Furthermore, a modified thiol degradation method using strongly acidic conditions was applied to the extract to yield three thiol degradation products derived from robinetinidol units. This method is useful for characterizing acacia proanthocyanidins (wattle tannins).