LC/MS analysis of stratum corneum lipids: ceramide profiling and discovery[S]

• Published in: Journal of lipid research Vol. 52; no. 6; pp. 1211 - 1221
• Main Authors: van Smeden, Jeroen, Hoppel, Louise, van der Heijden, Rob, Hankemeier, Thomas, Vreeken, Rob J., Bouwstra, Joke A.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01-06-2011; The American Society for Biochemistry and Molecular Biology; Elsevier
• Subjects: 3D multi-mass chromatogram; Adult; atmospheric pressure chemical ionization mass spectrometry; Ceramides - analysis; Ceramides - chemistry; Ceramides - classification; Ceramides - isolation & purification; Chromatography, Liquid - methods; human skin lipids; Humans; identification; Lipid Metabolism - physiology; liquid chromatography; Male; mass spectrometry; Mass Spectrometry - methods; normal-phase liquid chromatography; Skin - chemistry; Specimen Handling - methods; Spectroscopy, Fourier Transform Infrared; sphingolipids; Tissue Culture Techniques; human skin lipids; identification; 3D multi-mass chromatogram; sphingolipids; normal-phase liquid chromatography; liquid chromatography; mass spectrometry; atmospheric pressure chemical ionization mass spectrometry
• ISSN: 0022-2275; 1539-7262
• DOI: 10.1194/jlr.M014456

Ceramides (CERs) in the upper layer of the skin, the stratum corneum (SC), play a key role in the skin barrier function. In human SC, the literature currently reports 11 CER subclasses that have been identified. In this paper, a novel quick and robust LC/MS method is presented that allows the separation and analysis of all known human SC CER subclasses using only limited sample preparation. Besides all 11 known and identified subclasses, a 3D multi-mass chromatogram shows the presence of other lipid subclasses. Using LC/MS/MS with an ion trap (IT) system, a Fourier transform-ion cyclotron resonance system, and a triple quadrupole system, we were able to identify one of these lipid subclasses as a new CER subclass: the ester-linked ω-hydroxy fatty acid with a dihydrosphingosine base (CER [EOdS]). Besides the identification of a new CER subclass, this paper also describes the applicability and robustness of the developed LC/MS method by analyzing three (biological) SC samples: SC from human dermatomed skin, human SC obtained by tape stripping, and SC from full-thickness skin explants. All three biological samples showed all known CER subclasses and slight differences were observed in CER profile.