Individual variation of human plantar stratum corneum lipids, determined by automated multiple development of high-performance thin-layer chromatography plates |
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Authors: | Sebastian Zellmer,Jü rgen Lasch |
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Affiliation: | Institut für Physiologische Chemie, Martin-Luther-Universität Halle-Wittenberg, D-06097 Halle, Germany |
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Abstract: | The stratum corneum lipids are unique in composition and have been used frequently as a model system of the skin's lipid barrier. Automated multiple development (AMD) of high-performance thin-layer chromatography plates in combination with a 25-step gradient, based on methanol, diethyl ether and n-hexane separated the six major human plantar stratum corneum lipids. Post-chromatographic staining of these lipids with a solution of MnCl2H2SO4 at 130°C or a solution of CuSO4H3PO4 at 140°C allowed visualization of the lipids and quantification. The MnCl2H2SO4 solution stained saturated fatty acids less intensity. Therefore, the CuSO4H3PO4 solution was used for quantification and we found, on average, 2.06% (w/w) cholesterol 3-sulphate, 20.16% (w/w) free fatty acids, 20.25% (w/w) ceramides, 43.53% (w/w) non-esterified sterols, 4.56% (w/w) triacylglycerols and 9.4% (w/w) sterolesters in the human plantar stratum corneum extracts. The concentration of phospholipids was less than 1% (w/w). In addition, the lipid composition of twenty different human plantar stratum corneum extracts was determined. Statistics revealed a correlation between the ratio of free fatty acids and non-esterified sterols (r=0.832, p<0.01, n=20). Several control experiments proved that this correlation is not due to the extraction method, the post-chromatographic staining procedure or bacterial contamination of the stratum corneum. |
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Keywords: | Stratum corneum Automated multiple development Lipids Fatty acids |
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