<jats:title>Abstract</jats:title><jats:p>The wood and bark of four <jats:italic>Acacia</jats:italic> species growing in Portugal, namely, <jats:italic>A. longifolia, A. dealbata, A. melanoxylon</jats:italic>, and <jats:italic>A. retinodes</jats:italic>, were investigated for their sterol content. The lipids fractions of the different wood and bark samples were isolated, and the sterols were identified and quantified by GC‐MS. Two Δ<jats:sup>7</jats:sup> sterols, specifically, spinasterol and dihydrospinasterol, were the main sterols found in considerable amounts, particularly in wood tissues (more than 0.5 g/kg of dry wood in the case of <jats:italic>A. melanoxylon</jats:italic> and <jats:italic>A. retinodes</jats:italic>). The corresponding unusual steryl glucosides were also identified in significant amounts in the wood and bark extracts.