Contribution to Liquid Chromatographic Analysis of Cutaneous Ceramides

Gaudin, Karen; Chaminade, Pierre; Baillet, A.; Ferrier, D.; Bleton, Jean; Goursaud, S.; Tchapla, Alain

doi:10.1081/jlc-100101667

Cited by 37 publications

(20 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The other peaks did not correspond to any commercial reference substances. The lack of reference substances led us to establish a relationship between the structure and the retention behaviour of ceramides to build a retention model which constitutes the first tool for ceramide identification [25].…”

Section: A P P L I C a T I O N Smentioning

confidence: 99%

See 1 more Smart Citation

Analysis of commercial ceramides by non-aqueous reversed-phase liquid chromatography with evaporative light-scattering detection

et al. 1999

Self Cite

View full text Add to dashboard Cite

SummaryThis paper describes the development of a chromatographic system for analysis of commercial ceramides structurally similar to those found in the stratum corneum. The ceramides used in this study contain different amine bases (phytosphingosine, sphingosine and dihydrosphingosine) and fatty acids of different chain lengths and with different functional groups (hydroxylated and unsaturated). Non-aqueous reversed-phase (NARP) liquid chromatography with evaporative light-scattering detection (ELSD) were the techniques chosen in accordance with the nature of the ceramides. The eluent strength and the potential selectivity of different organic solvents were investigated. On a C18-bonded silica, the most promising chromatographic conditions employed a gradient from ACN-THF, 95:5, to ACN-THF-PrOH, 35:5:60, in 15 rain with a constant concentration of TEA (10 mM) and a stoichiometric amount of formic acid.

show abstract

Section: A P P L I C a T I O N Smentioning

confidence: 99%

“…To enable study these structures, the final part of our study was devoted to optimization of detection. The compounds present in each commercial standard (the major peak and the trace homologues) were individually characterized by gas chromatography coupled with mass spectrometry after methanolysis [25].…”

Section: Optimization Of Detectionmentioning

confidence: 99%

Analysis of commercial ceramides by non-aqueous reversed-phase liquid chromatography with evaporative light-scattering detection

et al. 1999

Self Cite

View full text Add to dashboard Cite

show abstract

“…In our laboratory, several methods for separation and structural identification of ceramide molecular species were developed [7,4,28]. One of these methods using PGC and methanol/chloroform gradient elution [7] was transposed in capillary-LC.…”

Section: Analysis Of Ceramide Molecular Species In Capillary-lcmentioning

confidence: 99%

“…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.…”

mentioning

confidence: 99%

Microanalytical systems for separations of stratum corneum ceramides

et al. 2006

View full text Add to dashboard Cite

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...

show abstract

“…Other studies presented methods to quantify SL using high-performance thin layer chromatography (HPTLC) and fluorescence spectrodensitometry (13) or reverse phase chromatography and GC-MS (14). Even though some of these methods can be used to determine the molecular species of free Cer or Cer moieties, they all require derivatizations, which not only cause the partial degradation of SL but also are time-consuming.…”

Section: Introductionmentioning

confidence: 99%

HPLC Quantification of Sphingolipids in Soybeans with Modified Palmitate Content

Wang

Fehr

2006

J. Agric. Food Chem.

View full text Add to dashboard Cite

Efficient separation and accurate quantification of sphingolipids (SL) are important for studying SL concentrations and biological functions. The objectives of this study were to develop effective methods for the separation and quantification of SL and to determine the relationship between palmitate and SL contents of mature soybean seeds. Methods using column chromatography and high-performance liquid chromatography−evaporative light scattering detector (HPLC-ELSD) were developed to separate and quantify glucosylceramide (GlcCer) and ceramide (Cer) in 15 soybeans lines in which palmitate content ranged from 3.7 to 40.7%. There were significant differences among the lines for GlcCer (83.4−397.6 nmol/g) and major Cer contents (8.4−20.7 nmol/g) on a dry weight basis. The correlations of palmitate content with GlcCer and Cer concentrations were not significant. The results indicated that the palmitate content of soybean seed did not affect their GlcCer and Cer contents. Genetic factors other than those that control palmitate content seemed to be responsible for the variation among soybean lines for GlcCer and Cer contents. Efficient separation and accurate quantification of sphingolipids (SL) are important for studying SL concentrations and biological functions. The objectives of this study were to develop effective methods for the separation and quantification of SL and to determine the relationship between palmitate and SL contents of mature soybean seeds. Methods using column chromatography and high-performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) were developed to separate and quantify glucosylceramide (GlcCer) and ceramide (Cer) in 15 soybeans lines in which palmitate content ranged from 3.7 to 40.7%. There were significant differences among the lines for GlcCer (83.4-397.6 nmol/g) and major Cer contents (8.4-20.7 nmol/g) on a dry weight basis. The correlations of palmitate content with GlcCer and Cer concentrations were not significant. The results indicated that the palmitate content of soybean seed did not affect their GlcCer and Cer contents. Genetic factors other than those that control palmitate content seemed to be responsible for the variation among soybean lines for GlcCer and Cer contents.

show abstract

Contribution to Liquid Chromatographic Analysis of Cutaneous Ceramides

Cited by 37 publications

References 25 publications

Analysis of commercial ceramides by non-aqueous reversed-phase liquid chromatography with evaporative light-scattering detection

Analysis of commercial ceramides by non-aqueous reversed-phase liquid chromatography with evaporative light-scattering detection

Microanalytical systems for separations of stratum corneum ceramides

HPLC Quantification of Sphingolipids in Soybeans with Modified Palmitate Content

Contact Info

Product

Resources

About