Plant seeds are arenewable resource that can furnish access tomedicinal natural products that can only otherwise be isolated fromaerial or root parts, the harvest of which may be destructive to theplant or threaten its viability. However, optimization of the isolationof such compounds from seeds would be greatly assisted if the spatialdistribution of the molecules of interest within the plant tissuewere known. For example, iboga alkaloids that hold promise for thetreatment of opioid use disorder are typically isolated from the leaves,bark, or roots of Tabernanthe or Voacanga spp. trees, but it would be more environmentally sustainable toisolate such compounds from their seeds. Here, we leveraged the uniquecapabilities of the ambient mass spectral imaging technique termedlaser ablation direct analysis in real-time imaging-mass spectrometry(LADI-MS) to reveal the spatial distributions of a range of molecules,including alkaloids within V. africana seeds. In addition to six compounds previously reported in theseseeds, namely, tetradecanoic acid, n-hexadecanoicacid, (Z,Z)-9,12-octadecadienoicacid, (Z)-9-octadecenoic acid, octadecanoic acid,and & UDelta;(14)-vincamine, an additional 31 compounds werenewly identified in V. africana seeds.The compound classes included alkaloids, terpenes, and fatty acids.The ion images showed that the fatty acids were localized in the embryoof the seed. The alkaloids, which were mainly localized in the seedendosperm, included strictamine, akuammidine, polyneruidine, vobasine,and & UDelta;(14)-vincamine. This information can be exploitedto enhance the efficiency of secondary metabolite isolation from V. africana seeds while eliminating the destructionof other plant parts.