Progress in the analysis of Stratum corneum ceramides

Raith, Klaus; Farwanah, Hany; Wartewig, S.; Neubert, Reinhard H.H.

doi:10.1002/ejlt.200400982

Cited by 28 publications

(25 citation statements)

References 121 publications

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“…The structural differences between these classes are the presence of a carbon double bond, two hydroxyl groups on the base moiety and one hydroxyl group on the fatty acid moiety in case of Cer 5 whereas Cer 3 possesses three hydroxyl groups on the sphingoid base moiety. Such behaviour was already observed in previous studies using different stationary phases [16,19].…”

Section: Solvent Selection With Pva-sil Stationary Phasesupporting

confidence: 85%

“…lipid class separation [3,9,14], micro-LC was envisaged in order to take advantage of the lower LOD expected with this technique [15] and its possible hyphenation with MS [16].…”

Section: Introductionmentioning

confidence: 99%

“…Normal-phase-HPLC (NP-HPLC) [11,16,17] or supercritical [18] methods were developed for SC lipid class analysis with conventional chromatographic columns. Silica gel remains the most used stationary phase in lipid class analysis [16,19].…”

Section: Introductionmentioning

confidence: 99%

“…Silica gel remains the most used stationary phase in lipid class analysis [16,19]. The heterogeneous surface of silica gel leads to variable adsorptive properties, irreversible adsorption of solutes and long reconditioning time.…”

Section: Introductionmentioning

confidence: 99%

“…As the evaporative light scattering detection (ELSD) is compatible with gradient procedure and the use of organic mobile phase, it has already demonstrated its suitability for ceramide analysis [16,17,19,21]. Furthermore when it is hyphenated with micro-LC, lower LOD can be reached [6] and ELSD leads eventually to more linear response for quantification study [24].…”

Section: Introductionmentioning

confidence: 99%

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Microanalytical systems for separations of stratum corneum ceramides

et al. 2006

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Original PaperMicroanalytical systems for separations of stratum corneum ceramidesThe small amount of lipids from human skin obtained with noninvasive sampling method led us to investigate microanalytical separation techniques. The lipid class analysis was performed with a micro polyvinyl alcohol-silica (PVA-Silm) column. The gradient elution was from heptane to acetone/butanol 90:10 v/v in 4%/min at 78 lL/ min. In addition an evaporative light scattering detector (ELSD) was modified for micro-LC. All solvents contained 0.1% of triethylamine and formic acid in stoichiometric amount, which increased the ELSD response. In these conditions, the cholesterol eluted before free fatty acid, and squalene and triglycerides close to the dead volume. The various ceramide classes eluted following the order of the increased number of hydroxyl groups. The LOD for ceramides was 2.2 ng. The advantages of this method are the use of a normal stationary phase more reliable due to its chemical stability, its surface homogeneity and its development in microchromatography without chlorinated solvents which offers small LOD and the whole profile of lipids present in stratum corneum (SC). A method using a narrow-bore PVA-Sil column was used to collect ceramide fraction. Then the molecular species were analysed with a porous graphitic carbon column in capillary LC using a gradient from CH 3 OH/CHCl 3 70:30 v/v to CHCl 3 at 2%/min with a flow rate at 5 lL/min. The LOD obtained for ceramide was 1 ng. Both methods were assessed with SC samples obtained by rinsing a 5.7 cm 2 area of the forearm with 25 mL of ethanol. IntroductionStratum corneum (SC) consists of corneocytes organized in a matrix of highly ordered multilamellar lipid sheets [1,2]. For the structural and physical properties of this lipid arrangement, SC is considered to play an essential role in the skin barrier function. Ceramides constitute 50% of this lipid barrier. Free fatty acid and cholesterol (Cho) constitute the remaining lipid classes composing this lipid barrier. The ceramide classes are highly heterogeneous in structure due to the sphingoid base and fatty acid moieties constituting these amide molecules. They differ by a different number of hydroxyl groups and carbon double bonds, and the chain length [3]. This structural heterogeneity is an important element to study the lipid barrier property. The nature of the structural difference between molecular species included in each lipid class is apolar. Therefore, apolar stationary phases are required for their discrimination such as octadecyl bonded silica [4][5][6] or porous graphitic carbon (PGC) [7]. This latter column provides original selectivity between molecular species. However, to apply such analytical methods to skin samples, the lipids should be previously extracted and the various lipid classes separated.Several noninvasive methods for extraction of lipids from SC exist, e.g. by stripping [8][9][10], scraping [11], skin rinsing [3,12] or with an extraction chamber [11,13]. Whatever the approach, the three...

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Section: Solvent Selection With Pva-sil Stationary Phasesupporting

confidence: 85%

“…lipid class separation [3,9,14], micro-LC was envisaged in order to take advantage of the lower LOD expected with this technique [15] and its possible hyphenation with MS [16].…”

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Microanalytical systems for separations of stratum corneum ceramides

et al. 2006

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The Importance of Stratum Corneum Lipid Organization for Proper Barrier Function

Schroeter

Eichner

Mueller

et al. 2015

Percutaneous Penetration Enhancers Chemical Methods in Penetration Enhancement

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

et al. 2005

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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 omega-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 omega-esterified ceramides. In addition, evidence for a new ceramide class Cer [NdS] was found.

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Progress in the analysis of Stratum corneum ceramides

Cited by 28 publications

References 121 publications

Microanalytical systems for separations of stratum corneum ceramides

Microanalytical systems for separations of stratum corneum ceramides

The Importance of Stratum Corneum Lipid Organization for Proper Barrier Function

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

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