Separation and mass spectrometric characterization of covalently bound skin ceramides using LC/APCI-MS and Nano-ESI-MS/MS.
Ceramides covalently bound to keratinocytes are essential for the barrier function of the skin, which can be disturbed in diseases, such as psoriasis and atopic dermatitis. These ceramides of the classes omega-hydroxyacyl-sphingosine and omega-hydroxyacyl-6-hydroxysphingosine contain an omega-hydroxy fatty acid. For their separation and identification, a new analytical approach based on normal phase liquid chromatography coupled to atmospheric pressure chemical ionization mass spectrometry and tandem nano-electrospray mass spectrometry, respectively, is presented here. Tandem mass spectrometry provided structural information about the sphingoid base as well as the fatty acid moieties. The chain lengths of the bases ranged from C12 to C22, the chain lengths of the fatty acids varied between C28 and C36. In total, 67 ceramide species have been identified in human skin. The analytical methods presented in this work can be helpful for investigating alterations in the ceramide composition of the skin as seen in psoriasis, atopic dermatitis, and diseases with impaired epidermal barrier function.
Farwanah
Hany
H
LIMES-Life and Medical Sciences Bonn, Program Unit Membrane and Lipid Biochemistry, Laboratory of Biology & Lipid Biochemistry, Friedrichs-Wilhelm-University, Gerhard-Domagk-Strasse 1, D-53121 Bonn, Germany.
Pierstorff
Barbara
B
Schmelzer
Christian E H
CE
Raith
Klaus
K
Neubert
Reinhard H H
RH
Kolter
Thomas
T
Sandhoff
Konrad
K
D001274:Atmospheric Pressure; D002518:Ceramides / Q000032:analysis / Q000302:isolation & purification*; D002853:Chromatography, Liquid / Q000379:methods*; D002855:Chromatography, Thin Layer; D003720:Densitometry; D013058:Mass Spectrometry / Q000379:methods*; D036103:Nanotechnology; D012867:Skin / Q000737:chemistry*
D002518:Ceramides