Isolation of minor compounds from natural extracts remains a complicated task and development of new protocols to facilitate the access to new structure is still needed. Application of a highly efficient 2D LC hyphenated to high resolution mass spectroscopy on the crude extract of the Mediterranean marine sponge Crambe crambe led to the identification of seven new compounds. The structure elucidation has been performed using 2D LC-MS/MS and NMR. The structure elucidation of these minor metabolites encouraged us to speculate on the metabolic pathway leading to this important family of sponge alkaloids. (C) 2014 Phytochemical Society of Europe. Published by Elsevier B.V. All rights reserved.