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Comparative investigation of human stratum corneum ceramides

  • Published:
Lipids

Abstract

The stratum corneum (SC) requires ceramides, cholesterol, and fatty acids to provide the cutaneous permeability barrier. SC lipids can be analyzed by normal-phase high-performance thin-layer chromatography (HPTLC). However, without further analysis, some uncertainty remains about the molecular composition of lipids represented by every TLC band of an unknown sample. We therefore analyzed each ceramide band further by subjecting the isolated lipids to a direct coupling of reversed-phase high-performance liquid chromatography and electrospray ionization-mass spectrometry (HPLC/ESI-MS, or IC/MS). IC/MS analysis and ESI-MS/MS negative ion and collision-induced dissociation experiments revealed that ceramide band 4 contained not only N-(ω-OH-acyl)acyl-6-OH-sphingosine, Cer(EOH), but also N-(α-OH-acyl)-sphingosine. Band 5 exclusively contained N-acyl-6-OH-sphingosine. Our results demonstrate the benefit of LC/MS analysis for selective identification of human SC ceramides. Moreover, the combination of HPTLC for pre-separation and LC/MS for identification of lipids is an even more powerful tool for detailed ceramide analysis.

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Abbreviations

Cer(AP):

N-(α-OH-acyl)-4-OH-sphinganine

Cer(AS):

N-(α-OH-acyl)-sphingosine

Cer(EOH):

N-(ω-OH-acyl)acyl-6-OH-sphingosine

Cer(EOP):

an ω-esterified ceramide containing phytosphingosine as a sphingosine backbone and an ω-esterified linoleic acid

Cer(EOS):

N-(ω-OH-acyl)acyl-sphinganine

Cer(NH):

N-acyl-6-OH-sphingosine

Cer(NP):

N-acyl-4-OH-sphinganine

Cer(NS):

N-acyl-sphingosine

CID:

collision-induced dissociation

ELSD:

evaporative light-scattering detection

ESI-MS:

electrospray ionization—mass spectrometry

HPLC:

high-performance liquid chromatography

HPTLC:

high-performance thin-layer chromatography

LC-MS:

liquid chromatography-mass spectrometry

MS:

mass spectrometry

MS/MS:

mass spectrometric fragmentation of specific ions

NMR:

nuclear magnetic resonance

RP:

reversed-phase

SC:

stratum corneum

TLC:

thin-layer chromatography

UV:

ultraviolet

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Authors and Affiliations

  1. Department of Dermatology and Allergy, Technical University of Munich, 80802, München, Germany

    Oliver Brandt & Dietrich Abeck

  2. Beiersdorf AG, Fkt. 4231, Unnastrasse 48, 20245, Hamburg, Germany

    Jens-Peter Vietzke, Claudius Rapp, Manfred Strassner, Volker Schreiner & Ulrich Hintze

Authors
  1. Jens-Peter Vietzke
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  2. Oliver Brandt
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  3. Dietrich Abeck
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  4. Claudius Rapp
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  7. Ulrich Hintze
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Correspondence to Jens-Peter Vietzke.

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Vietzke, JP., Brandt, O., Abeck, D. et al. Comparative investigation of human stratum corneum ceramides. Lipids 36, 299–304 (2001). https://doi.org/10.1007/s11745-001-0721-9

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  • DOI: https://doi.org/10.1007/s11745-001-0721-9

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