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ANANBIOANAL - 2010
Pharmaceutical R & D Summit
doi:10.4172/2155-9872.1000079
Investigation of the Detailed Molecular Structure of the Human
Stratum Corneum Ceramides [NP] and [EOS] by APCI and Nano-
ESI Mass Spectrometry
Reinhard H. H. Neubert
Institute of Pharmacy, Martin Luther University Halle-Wittenberg, Halle (Saale), Germany
T
he aim of this study was to characterize two ceramide subclasses, CER[NP]
and CER[EOS], of human stratum corneum (SC) and to identify the chemical
structures of their subspecies. High performance thin layer chromatography (HPTLC)
and normal phase high performance liquid chromatography (NP-HPLC) were used
for the separation of CER fractions of interest, whereas nanoelectrospray tandem
mass spectrometry was applied to identify the chemical structures in detail. Thus,
CER[EOS] fragmentation revealed that in addition to linoleic acid other esterified
fatty acids occur in the ω-hydroxylated-position (part E). Of particular interest is the
identification of a 17:2 fatty acid located in this part of the molecule.
Several subspecies of CER[NP], including subspecies with odd numbers of carbon
atoms in both chains, the non-α-hydroxylated fatty acid moiety (part N) and the
phytosphingosine (part P) were detected. Furthermore, 12% of CER[NP] subspecies
with an odd number of carbon atoms in more than one chain for one molecule were
detected.
Similar results were obtained for CER[EOS]. Both, the esterified fatty acid (part
E) and the sphingosine base (part S) were found to contain odd-numbered chain
lengths.
The combination of the analytical techniques presented allows complete new
insights into the molecular structure of the SC ceramides. Now the techniques are
used to identify differences in the detailed molecular structure of the ceramides in
healthy and diseased skin.
ANALBIOANAL-2010
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