Profiling of human stratum corneum ceramides by means of normal phase LC/APCI�MS

Farwanah, Hany; Wohlrab, Johannes; Neubert, Reinhard H. H.; Raith, Klaus
October 2005
Analytical & Bioanalytical Chemistry;Oct2005, Vol. 383 Issue 4, p632
Academic Journal
The ceramides of the stratum corneum are critical to maintaining the epidermal barrier function of the skin. A number of skin diseases and disorders are known to be related to impairments of the ceramide pattern. Therefore, obtaining mass spectrometric profiles of the nine ceramide classes known to exist aids our understanding of the underlying molecular mechanisms, which should eventually lead to new diagnostic opportunities: for example, the mass spectrometric profiles of patients suffering from serious skin diseases such as atopic dermatitis and psoriasis can be compared to those of healthy controls. Previous work on mass spectrometric analysis of ceramides relied mostly on GC/MS after hydrolysis and derivatization. The introduction of ESI�MS and LC/ESI�MS has provided new options for directly analyzing intact ceramides. However, some of the ceramide classes are not accessible to ESI�MS. However, as shown in this work, these limitations of GC/MS and ESI-MS can be overcome using a new approach based on normal phase LC interfaced with APCI�MS. Separation and online detection of the stratum corneum ceramide classes became possible in one run. Ceramide species with C26 and/or C28 fatty acid chains were the most abundant ones in Cer [NP], Cer [NH], Cer [AP], and Cer [AH]. The main component of Cer [AS] was C16. The ?-esterified ceramide classes Cer [EOS], Cer [EOP] and Cer [EOH] contained mostly species with fatty acids >C30. This was also the case for Cer [NS], suggesting an analogy to the ?-esterified ceramides. In addition, evidence for a new ceramide class Cer [NdS] was found.


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