Electron Paramagnetic Resonance Investigation of Stratum Corneum Lipid Structure

Electron paramagnetic resonance (EPR) in conjunction with a slow-tumbling simulation was utilized for defining stratum corneum (SC) lipid structure. We found that ordering calculated from the simulation is an appropriate index for evaluating SC lipids structure. The SC from two sites (mid-volar forearm and lower-leg) of human volunteers was stripped consecutively from one to three times using a glass plate coated with a cyanoacrylate resin. Aliphatic spin probes, 5-doxylstearic acid (5-DSA) and 3β-doxyl-5α-cholestane (CHL), were used to monitor SC ordering. EPR spectrum of 5-DSA incorporated in the SC demonstrated a characteristic peak for the first strip. However, EPR spectra of CHL in the SC did not show a clear difference for each strip, except for the peak intensity. The results imply that CHL is not incorporated into the lipid phase as easily as 5-DSA is. A slow-tumbling simulation of the EPR spectrum was performed to analyze the detailed lipid structure. The simulation results for 5-DSA show differences in values of the SC ordering as a function of depth. Thus, these results along with the simulation analysis provide a detailed SC layer structure.