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Papers In Press, published online ahead of print July 1, 2008
J. Lipid Res., doi:10.1194/jlr.M800014-JLR200

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Journal of Lipid Research, Vol. 49, 1466-1476, July 2008
Copyright © 2008 by American Society for Biochemistry and Molecular Biology

Characterization of overall ceramide species in human stratum corneum

Yoshinori Masukawa¹, Hirofumi Narita, Eri Shimizu, Naoki Kondo, Yoshiya Sugai, Tsuyoshi Oba, Rika Homma, Junko Ishikawa, Yutaka Takagi, Takashi Kitahara, Yoshinori Takema and Katsumi Kita

Tochigi Research Laboratories, Kao Corporation, Ichikai, Haga, Tochigi 321-3497, Japan

The online version of this article (available at http://www.jlr.org) contains supplementary data in the form of two figures and two tables.

Published, JLR Papers in Press, March 23, 2008.

¹ To whom correspondence should be addressed. e-mail: masukawa.yoshinori{at}kao.co.jp

Ceramides (CERs) in human stratum corneum (SC) play physicochemical roles in determining barrier and water-holding functions of the skin, and specific species might be closely related to the regulation of keratinization, together with other CER-related lipids. Structures of those diverse CER species, however, have not been comprehensively revealed. The aim of this study was to characterize overall CER species in the SC. First, we constructed 3D multi-mass chromatograms of the overall CER species, based on normal-phase liquid chromatography (NPLC) connected to electrospray ionization-mass spectrometry (ESI-MS) using a gradient elution system and a postcolumn addition of a volatile salt-containing polar solvent. The CERs targeted from the 3D chromatograms were structurally analyzed using NPLC-ESI-tandem mass spectrometry (MS/MS), which resulted in the identification of 342 CER species in the inner forearm SC. This led to the discovery of a new CER class consisting of -hydroxy fatty acid and dihydrosphingosine moieties, in addition to the 10 classes generally known. The results also revealed that those CERs contain long-chain (more than C₁₈)-containing sphingoids and a great number of isobaric species. These novel results will contribute not only to physiochemical research on CERs in the SC but also to lipidomics approaches to CERs in the skin.

Supplementary key words electrospray ionization tandem mass spectrometry • identification • isobaric • multi-mass chromatograms • normal-phase liquid chromatography • long chain-containing sphingoid

Abbreviations: CER, ceramide; ESI-MS, electrospray ionization-mass spectrometry; MS/MS, tandem mass spectrometry; NPLC, normal-phase liquid chromatography; SC, stratum corneum; TIC, total ion chromatogram

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