Separation and identification of ceramides in the human stratum corneum by high-performance liquid chromatography coupled with electrospray ionization mass spectrometry and electrospray multiple-stage mass spectrometry profiling

Vietzke, Jens-Peter; Straßner, M.; Hintze, Ulrich

doi:10.1007/bf02493611

Cited by 26 publications

(21 citation statements)

References 26 publications

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“…An advantage of ion traps is their capability to perform multiple stage MS (MS n ), which allows for structure elucidation by stepwise fragmentation, provided that the software is able to control MS n . This was also demonstrated for stratum corneum ceramides by Vietzke et al [75]. A number of other papers described the successful use of ESI-MS/MS to characterize ceramides in biological samples [83][84][85][86][87][88].…”

Section: Mass Spectrometrymentioning

confidence: 68%

Progress in the analysis of Stratum corneum ceramides

Raith

Farwanah

Wartewig

et al. 2004

Euro J Lipid Sci & Tech

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Progress in the analysis of Stratum corneum ceramidesThe stratum corneum with its unique structure of corneocytes and intercellular lipid lamellae enables a protection of human and other mammalian skin against transepidermal water loss and harmful substances from the environment. Among these lipids, ceramides play a key role. Several skin disorders such as psoriasis, atopic dermatitis and others show a disturbed barrier function, which can be linked in part to a changed ceramide pattern. Research in dermatology and cosmetic industry requires the use of analytical methods to characterize ceramides. However, the amazing variability of ceramide structures found in stratum corneum makes their analysis a challenge. This is to our knowledge the first review that deals with ceramide analysis. It is focused on stratum corneum ceramides which show much more complexity than intracellular ceramides known to be signal transducers. After a short summary of lipid extraction methods, we discuss the following set of methods: thin-layer chromatography, gas chromatography, HPLC, LC/MS, mass spectrometry, NMR spectroscopy, vibrational spectroscopy, and X-ray diffraction. It will be lined out which information about structure, concentration or physical state is available by means of each particular method. Although the focus is clearly on stratum corneum ceramides, some methods of general skin lipid analysis and structure analysis of sphingolipids are also discussed when appropriate.

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Section: Mass Spectrometrymentioning

confidence: 68%

Progress in the analysis of Stratum corneum ceramides

Raith

Farwanah

Wartewig

et al. 2004

Euro J Lipid Sci & Tech

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“…To analyze CERs in human SC using LC-MS, we decided to use NP and ESI modes in LC and MS, respectively, because the NP mode may enable the class separation of diverse CERs that have chain distributions both in the fatty acid and sphingoid moieties based on differences in their polarities, whereas the ESI mode seems to be the best for the LC-MS analysis of CERs, as demonstrated in many analytical studies (14,15,(32)(33)(34)(35)(36).…”

Section: Optimization Of Nplc-esi-ms Conditionsmentioning

confidence: 99%

Characterization of overall ceramide species in human stratum corneum

Masukawa

Narita

Shimizu

et al. 2008

Journal of Lipid Research

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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“…This indicates However, there seemed to be no tendency that specifi c fatty acid or sphingoid moieties are related to the greater RSD. This might be related to relatively unstable performances of NPLC-ESI-MS, especially on separate days, compared with RPLC-ESI-MS that is stable and commonly used in the analyses of CERs (22,(30)(31)(32). Relative response values are a ratio of the slope of the calibration curve (100-1000 fmol) each for 11 authentic CERs to that of the internal standard CER[N(17)S (18) (3) responses of isobaric species may be substantially similar to each other.…”

Section: Simultaneous Sim Measurement In Nplc-esi-msmentioning

confidence: 99%

“…To characterize differences in lipidomes among biological samples of interest, the relative intensities or areas given from mass spectrometers are only helpful as semi-quantitative values ( 11,12 ). Precise quantitative values of lipidomes of interest would also be desirable, especially in the case of evaluations of lipid dynamics in de MS has been employed for the analysis of CERs in human SC as follows: reversed-phase LC (RPLC)-ESI-MS ( 22,(30)(31)(32); ESI-MS ( 33,34 ); RPLC-thermospray-MS ( 35 ); and NPLC-APCI-MS (36)(37)(38)(39). However, those methods were not validated as quantitative methods for overall CER species in the SC, although some studies reported quantitative values based on unrealistic assumptions.…”

mentioning

confidence: 99%

Comprehensive quantification of ceramide species in human stratum corneum

Masukawa

Narita

Sato

et al. 2009

Journal of Lipid Research

209

150

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This article is available online at http://www.jlr.org novo pathways, such as alterations of some species of the targeted lipidomes into others. Similar to common quantitative analyses using direct MS or LC-MS that need authentic standards together with internal ones, quantitative lipidomics may also require a standard set of all lipid species targeted, because each lipid species has a different molar response in MS detection except for special cases. However, it is practically impossible to obtain all authentic species, including not only different even straight carbon chains but also odd and/or branched ones. To overcome such limitations, lipidomics researchers have contrived a novel procedure to quantify lipidomes comprehensively. Han and Gross ( 3 ) proposed a quantitative method for analyzing triglyceride molecular species using direct MS, which involves a procedure to calculate response factors for species that are not available as authentic materials, based on differences of the responses in the numbers of total carbons and double bonds among the species. For quantitative determination of phospholipid molecular species, Koivusalo et al. ( 4 ) utilized the following parameAbstract One of the key challenges in lipidomics is to quantify lipidomes of interest, as it is practically impossible to collect all authentic materials covering the targeted lipidomes. For diverse ceramides (CER) in human stratum corneum (SC) that play important physicochemical roles in the skin, we developed a novel method for quantifi cation of the overall CER species by improving our previously reported profi ling technique using normal-phase liquid chromatography-electrospray ionization-mass spectrometry (NPLC-ESI-MS). The use of simultaneous selected ion monitoring measurement of as many as 182 kinds of molecular-related ions enables the highly sensitive detection of the overall CER species, as they can be analyzed in only one SC-stripped tape as small as 5 mm × 10 mm. To comprehensively quantify CERs, including those not available as authentic species, we designed a procedure to estimate their levels using relative responses of representative authentic species covering the species targeted, considering the systematic error based on intra-/inter-day analyses. The CER levels obtained by this method were comparable to those determined by conventional thin-layer chromatography (TLC), which guarantees the validity of this method. , ceramide class consisting of non-hydroxy fatty acids and 4-sphingenines; ESI, electrospray ionization; LOD, limit of detection; LOQ, limit of quantifi cation; NPLC, normal-phase liquid chromatography; RPLC, reversed-phase liquid chromatography; RSD, relative standard deviation; SC, stratum corneum; SIM, selected ion monitoring; S/N, signal to noise; TLC, thinlayer chromatography

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Separation and identification of ceramides in the human stratum corneum by high-performance liquid chromatography coupled with electrospray ionization mass spectrometry and electrospray multiple-stage mass spectrometry profiling

Cited by 26 publications

References 26 publications

Progress in the analysis of Stratum corneum ceramides

Progress in the analysis of Stratum corneum ceramides

Characterization of overall ceramide species in human stratum corneum

Comprehensive quantification of ceramide species in human stratum corneum

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