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 identification and relative distribution of eleven classes of ceramides, which are separated by UHPLC and determined by their specific retention times. The precise composition 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 identified. 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 profiles between human and minipig stratum corneum were similar. Differences 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 measurement of the relative distribution of ceramides as biomarkers for pharmacodynamic studies.
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Acknowledgements
The authors are grateful to Charbel Azar and Thierry Alvarez for the collection of minipig stratum corneum samples, to Oana Hulea and Malthide Moyne for the development of UPLC methods, and to Paul Fogel and Michel Poncet for statistical assistance.
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Contributions
GPL, VA and AG performed all sample analysis. Data analysis and interpretation was carried out in collaboration between GPL, BG and JJV. GPL and JJV wrote the manuscript together. All authors have participed in critical data interpretation and acceptance of the manuscript.
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The authors declare that they have no conflict of interest and no conflict of financial interest.
Ethical approval
The work was performed in accordance with the code of ethics of the world medical association (Declaration of Helsinki). Informed consent was obtained for all 10 healthy volunteers enrolled in the study. The study was approved by the local ethics committee (CPP Sud-Méditerranée V, Hopital de Cimiez, NICE, France). For Göttingen minipigs, an equivalent sampling process was performed on the flank of 5 animals. All animal experiments were compliant with the EU Directive 2010/63/EU.
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Laffet, G.P., Genette, A., Gamboa, B. et al. Determination of fatty acid and sphingoid base composition of eleven ceramide subclasses in stratum corneum by UHPLC/scheduled-MRM. Metabolomics 14, 69 (2018). https://doi.org/10.1007/s11306-018-1366-4
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DOI: https://doi.org/10.1007/s11306-018-1366-4