Ceramides, a subclass of sphingolipids, are found in all tissues of the human body. The largest diversity is detected in human skin, more specifically in the uppermost layer of the epidermis, i.e. the stratum corneum, were ceramides play a prominent role in the barrier and permeability properties of the skin. Given the huge structural diversity, skin ceramide profiling requires the use of a highly efficient chromatographic step before mass spectrometric measurement. Micro pillar array columns (μPAC) are perfectly suited in that respect. Their unique benefits are illustrated in this application note.