New method to characterize epidermal Ceramides by Supercritical Fluid Chromatography (SFC) coupled to High Resolution Mass (Q-Tof) : Application to various samples with skin barrier default. - Inria - Institut national de recherche en sciences et technologies du numérique Accéder directement au contenu
Poster Année : 2023

New method to characterize epidermal Ceramides by Supercritical Fluid Chromatography (SFC) coupled to High Resolution Mass (Q-Tof) : Application to various samples with skin barrier default.

Résumé

Lipids are the main constituent of skin and epidermis, and are mainly constituted by ceramides with prevalent very long acyl chains, free fatty acids, and cholesterol. In health context, the lipid composition of skin can be highly modified in diseases thus its full characterization is essential. Epidermal ceramides, specificaly ω-O-Acylceramide, will have a crucial role for skin barrier function. The epidermal ceramides is a large and complex family of 12 sub-classes and their structural diversity, their wide range of concentration and the few standards available make this analysis very challenging. The aim of this work was to develop an original profiling of the epidermal ceramides using supercritical Fluid Chromatography (SFC) (UPC² Waters), which is a technology especially suitable for the non-polar metabolites, coupled to a Q-TOF (Xevo, Waters). The profiling was developed on a Torus DIOL column with a gradient of a mixture of isopropanol, methanol and acetonitrile (0.1% formic acid) in CO2 under pressure with ionisation in negative mode in presence of methanol. In studies involving the skin we can find many samples: it can be whole skin biopsies, cigarette paper impregnated with sebum, strip, solvent-in vivo skin extraction but also reconstructed human epidermis (RHE) for in vitro skin studies. Sample preparation needs to be adapted to sampling. We will present the development and the validation of this new method from the sample preparation to the data treatment to the chromatographic separation and structural characterisation. Mutation in patatin-like phospholipase domain-containing 1 (PNPLA1) causes autosomal recessive congenital ichthyosis. This mutation conducts to lethal phenotype with major defects in the epidermal barrier specially a blocage in ω-O-acylceramide synthesis. We will show the first application of the developed ceramide profiling on mutated Pnlpa1 mice and RHE samples.
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Dates et versions

hal-04245640 , version 1 (17-10-2023)

Identifiants

  • HAL Id : hal-04245640 , version 1

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Justine Bertrand-Michel, Cyrielle Clement, Pauline Le Faouder, Julia Soullier, Carla Orlandi, et al.. New method to characterize epidermal Ceramides by Supercritical Fluid Chromatography (SFC) coupled to High Resolution Mass (Q-Tof) : Application to various samples with skin barrier default.. Forum of lipidomics 2023, Aug 2023, Vienne (Austria), Austria. ⟨hal-04245640⟩
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