Liquid chromatography-mass spectrometry for comprehensive profiling of ceramide molecules in human hair

Masukawa, Yoshinori; Tsujimura, Hiroji; Narita, Hirofumi

doi:10.1194/jlr.d600007-jlr200

Cited by 42 publications

(62 citation statements)

References 48 publications

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“…5A ), very characteristic fragments were observed for both the sphingosine base as well as the fatty acid chain, which have also been reported in earlier studies ( 39,54,56,58 + ions, are also present in Fig. 5A .…”

Section: Because No Synthetic Variant Of Cer [Eods] Is Commercially Asupporting

confidence: 86%

“…This data was compared with that reported before ( 39,(53)(54)(55)(56)(57)(58). Following the synthetic CERs, CER fragment-ions of relevant human SC were analyzed.…”

Section: Because No Synthetic Variant Of Cer [Eods] Is Commercially Amentioning

confidence: 99%

“…5B . Again, fragments corresponding to the fatty acid chain and the sphingosine base were observed (54,56,58) TABLE 3. Overview of MS fragments of both synthetic and biological ceramides and its structural correlation Dotted lines with their corresponding numbers depict the location of fragmentation, viz.…”

Section: Ms/ms Of the Unknown Cer Classmentioning

confidence: 99%

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LC/MS analysis of stratum corneum lipids: ceramide profiling and discovery

Smeden

Hoppel²,

Heijden³

et al. 2011

Journal of Lipid Research

202

149

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organization of these lipids are of major importance for a competent skin barrier function ( 1-4 ). The major lipid classes in human SC are cholesterol, free fatty acids, and ceramides (CERs). In particular, the CERs have drawn much attention and several reported studies demonstrate that changes in CER composition may play a role in an impaired skin barrier ( 5-11 ). The various CERs consist of a sphingoid base linked via an amide bond to a fatty acid. Both variations in the fatty acid carbon chain and the sphingoid base architecture result in a large number of CER subclasses with a wide variation in chain length distribution. The molecular architecture of these subclasses is depicted in Fig. 1 . Over the years, important information about CER composition in human SC has been obtained using high performance thin layer chromatography (HPTLC) in conjunction with NMR. This resulted in a gradual increase in the number of identifi ed subclasses. By use of these methods, nine different human CER subclasses have been identifi ed in the SC of healthy human skin ( 12-16 ). As HPTLC is easily accessible, it is still frequently used in various fi elds with respect to skin lipid research to determine the CER subclasses in human SC ( 11,17,18 ). However, these approaches are very time consuming and a high quantity of material is required, which is not always available. The application of LC/MS results in a much more detailed profi le of each individual subclass using only small quantities of material (19)(20)(21)(22)(23)(24)(25)(26)(27)(28). Besides their important role in human skin, CERs are also key molecules with respect to cell signaling, growth, differentiation, and apoptosis ( 29-38 ). Also in these research areas, the introAbstract Ceramides (CERs) in the upper layer of the skin, the stratum corneum (SC), play a key role in the skin barrier function. In human SC, the literature currently reports 11 CER subclasses that have been identifi ed. In this paper, a novel quick and robust LC/MS method is presented that allows the separation and analysis of all known human SC CER subclasses using only limited sample preparation. Besides all 11 known and identifi ed subclasses, a 3D multimass chromatogram shows the presence of other lipid subclasses. Using LC/MS/MS with an ion trap (IT) system, a Fourier transform-ion cyclotron resonance system, and a triple quadrupole system, we were able to identify one of these lipid subclasses as a new CER subclass: the ester-linked -hydroxy fatty acid with a dihydrosphingosine base (CER [EOdS]). Besides the identifi cation of a new CER subclass, this paper also describes the applicability and robustness of the developed LC/MS method by analyzing three (biological) SC samples: SC from human dermatomed skin, human SC obtained by tape stripping, and SC from fullthickness skin explants. All three biological samples showed all known CER subclasses and slight differences were observed in CER profi le. The outermost layer of mammalian skin, the stratum corneum (SC), consists of corneocytes embedd...

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Section: Because No Synthetic Variant Of Cer [Eods] Is Commercially Asupporting

confidence: 86%

“…This data was compared with that reported before ( 39,(53)(54)(55)(56)(57)(58). Following the synthetic CERs, CER fragment-ions of relevant human SC were analyzed.…”

Section: Because No Synthetic Variant Of Cer [Eods] Is Commercially Amentioning

confidence: 99%

See 1 more Smart Citation

LC/MS analysis of stratum corneum lipids: ceramide profiling and discovery

Smeden

Hoppel²,

Heijden³

et al. 2011

Journal of Lipid Research

202

149

View full text Add to dashboard Cite

show abstract

“…Human hair CERs are more complex than are those in human cells/tissues, because hair contains 4 classes (combinations of nonhydroxy fatty acids and sphingosines, nonhydroxy fatty acids and dihydrosphingosines, a-hydroxy fatty acids and sphingosines, and a-hydroxy fatty acids and dihydrosphingosines), and the predominant class, CERs consisting of nonhydroxy fatty acid and dihydrosphingosine moieties, has C 16 -, C 17 -, C 18 -, C 19 -, or C 20 -containing dihydrosphingosines and not only even but also odd carbon-containing nonhydroxy fatty acids (16). On the other hand, human SC has even more-complex CERs.…”

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confidence: 99%

Characterization of overall ceramide species in human stratum corneum

Masukawa

Narita

Shimizu

et al. 2008

Journal of Lipid Research

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299

259

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Ceramides (CERs) in human stratum corneum (SC) play physicochemical roles in determining barrier and waterholding functions of the skin, and specific species might be closely related to the regulation of keratinization, together with other CER-related lipids. Structures of those diverse CER species, however, have not been comprehensively revealed. The aim of this study was to characterize overall CER species in the SC. First, we constructed 3D multi-mass chromatograms of the overall CER species, based on normalphase liquid chromatography (NPLC) connected to electrospray ionization-mass spectrometry (ESI-MS) using a gradient elution system and a postcolumn addition of a volatile saltcontaining polar solvent. The CERs targeted from the 3D chromatograms were structurally analyzed using NPLC-ESItandem mass spectrometry (MS/MS), which resulted in the identification of 342 CER species in the inner forearm SC. This led to the discovery of a new CER class consisting of ahydroxy fatty acid and dihydrosphingosine moieties, in addition to the 10 classes generally known. The results also revealed that those CERs contain long-chain (more than C 18 )-containing sphingoids and a great number of isobaric species. These novel results will contribute not only to physiochemical research on CERs in the SC but also to lipidomics approaches to CERs in the skin.-Masukawa, Y

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“…Hair includes ceramides that are predominantly composed of nonhydroxyl or α-hydroxyl fatty acids moiety and dihydrosphingosine moiety [7]. It was considered that ceramides may be related to apoptosis during keratinization that proceeds from living hair matrix cell to dead cuticular or cortical cells and may contribute to barrier function and water holding in hair [18]. It was established that both the free and covalently bound lipids composition of felted and felted yellowed wool fiber contains fewer of ceramides (P < 0.05) compared to undamaged fibers.…”

Section: Resultsmentioning

confidence: 99%