Individual variation of human plantar stratum corneum lipids, determined by automated multiple development of high-performance thin-layer chromatography plates.
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 MnCl2-H2SO4 at 130 degrees C or a solution of CuSO4-H3PO4 at 140 degrees C allowed visualization of the lipids and quantification. The MnCl2-H2SO4 solution stained saturated fatty acids less intensely. Therefore, the CuSO4-H3PO4 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.
Zellmer
S
S
Institut für Physiologische Chemie, Martin-Luther-Universität Halle-Wittenberg, Germany.
Lasch
J
J
D000818:Animals; D002712:Chlorides; D002855:Chromatography, Thin Layer / Q000295:instrumentation; D004396:Coloring Agents; D019327:Copper Sulfate; D005230:Fatty Acids, Nonesterified / Q000032:analysis; D005528:Foot; D006801:Humans; D008055:Lipids / Q000032:analysis* / Q000302:isolation & purification; D017895:Manganese Compounds; D010756:Phosphoric Acids; D015203:Reproducibility of Results; D012867:Skin / Q000737:chemistry*; D013261:Sterols / Q000032:analysis; D013464:Sulfuric Acids
D002712:Chlorides; D004396:Coloring Agents; D005230:Fatty Acids, Nonesterified; D008055:Lipids; D017895:Manganese Compounds; D010756:Phosphoric Acids; D013261:Sterols; D013464:Sulfuric Acids; C030242:phosphoric acid; D019327:Copper Sulfate; C033158:sulfuric acid; C025340:manganese chloride