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Article Dans Une Revue Physical Chemistry Chemical Physics Année : 2020

Quantification of the amount of mobile components in intact stratum corneum with natural-abundance 13C solid-state NMR

Résumé

The outermost layer of the skin is the stratum corneum (SC), which is mainly comprised of solid proteins and lipids. Minor amounts of mobile proteins and lipids are crucial for the macroscopic properties of the SC, including softness, elasticity and barrier function. Still this minor number of mobile components are not well characterized in terms of structure or amount. Conventional quantitative direct polarization (Q-DP) 13C solid-state NMR gives signal amplitudes proportional to concentrations, but fails to quantify the SC mobile components because of spectral overlap with the overwhelming signals from the solids. Spectral editing with the INEPT scheme suppresses the signals from solids, but also modulates the amplitudes of the mobile components depending on their values of the transverse relaxation times T2, scalar couplings JCH, and number of covalently bound hydrogens nH. This study describes a quantitative INEPT (Q-INEPT) method relying on systematic variation of the INEPT timing variables to estimate T2, JCH, nH, and amplitude for each of the resolved resonances from the mobile components. Q-INEPT is validated with a series of model systems containing molecules with different hydrophobicity and dynamics. For selected systems where Q-DP is applicable, the results of Q-INEPT and Q-DP are similar with respect to the linearity and uncertainty of the obtained molar ratios. Utilizing a reference compound with known concentration, we quantify the concentrations of mobile lipids and proteins within the mainly solid SC. By melting all lipids at high temperature, we obtain the total lipid concentration. These Q-INEPT results are the first steps towards a quantitative understanding of the relations between mobile component concentrations and SC macroscopic properties.
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hal-04292116 , version 1 (21-11-2023)

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Quoc Dat Pham, Goran Carlstrom, Olivier Lafon, Emma Sparr, Daniel Topgaard. Quantification of the amount of mobile components in intact stratum corneum with natural-abundance 13C solid-state NMR. Physical Chemistry Chemical Physics, 2020, Physical Chemistry Chemical Physics, 22, pp.6572-6583. ⟨10.1039/d0cp00079e⟩. ⟨hal-04292116⟩
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