Sphingolipids are important lipids and integral members of membranes, where they form small microdomains called lipid rafts, These rafts are enriched in cholesterol and sphingolipids, which influences biophysical properties. Interestingly, the membranes of the biomedical model organism Caenorhabditis elegans contain only low amounts of cholesterol. Sphingolipids in C. elegans are based on an unusual Cl7iso branched sphingoid base. In order to analyze and the sphingolipidome of C. elegans in more detail, we performed fractionation of lipid extracts and depletion of glycero- and glycerophospholipids together with in-depth analysis using UPLC-UHRToF-MS. In total we were able to detect 82 different sphingolipids from different classes, including several isomeric species.