Vol.:(0123456789) 1 3 Metabolomics (2018) 14:69 https://doi.org/10.1007/s11306-018-1366-4 ORIGINAL ARTICLE Determination of fatty acid and sphingoid base composition of eleven ceramide subclasses in stratum corneum by UHPLC/scheduled-MRM Gilbert P. Lafet 1  · Alexandre Genette 1  · Bastien Gamboa 1  · Virginie Auroy 2  · Johannes J. Voegel 1 Received: 10 January 2018 / Accepted: 2 May 2018 © Springer Science+Business Media, LLC, part of Springer Nature 2018 Abstract Introduction Ceramides play a key role in skin barrier function in homeostatic and pathological conditions and can be sampled non-invasively through stratum corneum collection. Objectives To develop a novel UHPLC/Scheduled MRM method for the identifcation and relative distribution of eleven classes of ceramides, which are separated by UHPLC and determined by their specifc retention times. The precise composi- tion of the fatty acid and sphingoid base parts of each individual ceramide is determined via mass fragmentation. Methods More than 1000 human and pig ceramides were identifed. Three human and minipig ceramide classes, CER[AS], CER[NS] and CER[EOS] have been investigated in depth. Results Sphingoid bases were characterized by a prevalence of chain lengths with sizes from C16 to C22, whereas fatty acids were mainly observed in the range of C22–C26. Overall, the ceramide profles between human and minipig stratum corneum were similar. Diferences in the CER[AS] and CER[NS] classes included a more homogeneous distribution of fatty acids (16–30 carbon atoms) in minipig, whereas in human longer fatty acid chains (> 24 carbon atoms) predominated. Conclusion The method will be useful for the analysis of healthy and pathological skin in various specie, and the measure- ment of the relative distribution of ceramides as biomarkers for pharmacodynamic studies. Keywords Ceramides · Stratum corneum · Non-invasive sampling · UHPLC/scheduled-MRM · Sphingoid bases · Fatty acids 1 Introduction The stratum corneum (SC) is the outermost layer of skin and the main barrier against transepidermal water and electrolyte loss. It protects the organism from toxic compounds, aller- gens and penetration of pathogenic microorganisms (Jun- gersted et al. 2008; Hinder et al. 2011; Feingold and Elias 2014; Elias 2014; Imokawa et al. 1991; Vietzke et al. 2001). The epidermal barrier function in human stratum corneum resides primarily in two compartments, enucleated corneo- cytes and extracellular lipids, which are often represented by the brick and mortar model. Skin is therefore a tissue highly specialized in the synthesis of lipids. Perturbed lipid metabolism leads to a disturbed skin barrier associated with difculties in maintaining fuid, electrolyte balance and a higher risk of infection. A relationship between altered SC lipid profles and impaired skin barrier function has been described in many skin diseases, such as atopic dermatitis (Kezic et al. 2014; Grösch et al. 2012; Elias 2014). The extracellular lipid matrix is composed of three main lipid classes: Ceramides (CER) form about 40–50% of the lipid content; cholesterol and cholesterol sulphate (CH and CS) form about 25% of lipid content and are involved in des- quamation and lipid organization and free fatty acids (FFA) form about 15% of lipid content and are mainly involved in lipid organization (Elias 2014; Feingold and Elias 2014; Janssens et al. 2012). Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11306-018-1366-4) contains supplementary material, which is available to authorized users. * Gilbert P. Lafet gilbert.lafet@galderma.com * Johannes J. Voegel johannes.voegel@galderma.com 1 Molecular Dermatology, Department of Research, Galderma R&D – Nestlé Skin Health, 2300 Route des Colles, 06902 Sophia Antipolis, France 2 Albhades Provence, 940 avenue de Traversetolo, 04700 Oraison, France