Simultaneous quantitative determination method for ceramide species from crude cellular extracts by high-performance liquid chromatography–thermospray mass spectrometry

Yamane, Mototeru

doi:10.1016/s1570-0232(02)00637-2

Cited by 14 publications

(12 citation statements)

References 26 publications

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“…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.…”

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confidence: 99%

Comprehensive quantification of ceramide species in human stratum corneum

Masukawa

Narita

Sato

et al. 2009

Journal of Lipid Research

211

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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confidence: 99%

Comprehensive quantification of ceramide species in human stratum corneum

Masukawa

Narita

Sato

et al. 2009

Journal of Lipid Research

211

150

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“…Although gas chromatography-mass spectrometry (MS) can individually detect trimethylsilylated CERs (23,24), it cannot comprehensively detect minor CERs as well as major CERs because of its lower sensitivity. Since Mano et al (29), Gu et al (30), and Couch et al (31) demonstrated MS using electrospray ionization (ESI) or atmospheric pressure chemical ionization (APCI) to be extremely valuable tools in the analysis of mixed CER molecules, many methods for the identification and determination of CER molecules have been proposed and applied to the analysis of CERs in human cells/tissues (10,(32)(33)(34)(35)(36)(37)(38)(39)(40)(41)(42)(43)(44)(45): reversed-phase liquid chromatography (RPLC) (34,35,39,40), normal-phase LC (41)(42)(43)45) connected to single quadrupole MS (39,40,44) or ion trap MS (34,35,(41)(42)(43)45), and direct triple quadrupole MS (10,32,33,(36)(37)(38). However, these methods do not provide comprehensive profiling of all CER molecules, including isobaric and isomeric CERs.…”

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confidence: 99%

“…However, these methods do not provide comprehensive profiling of all CER molecules, including isobaric and isomeric CERs. Furthermore, it is unknown whether those methods are optimal to detect lower levels of a-hydroxy FAM-containing CERs that are present in human cells/tissues, because those CERs were not detected in any of their applications (10,(32)(33)(34)(35)(36)(37)(38)(39)(40)(41)(42)(43)(44)(45), despite their successful detection in authentic samples.…”

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

Masukawa

Tsujimura

Narita

2006

Journal of Lipid Research

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Ceramides (CERs) play key roles in signal transduction and cell regulation, probably during the keratinization of human hair. Current methods using mass spectrometry (MS), however, are not sufficient to allow the comprehensive analysis of CER molecules, including isobaric and isomeric CERs. Therefore, a method for the comprehensive profiling of CERs was developed. The method developed is based on reversed-phase liquid chromatography (RPLC) coupled to atmospheric pressure chemical ionization (APCI)-MS. Comprehensive identification and profiling of CERs is achieved using two sets of multimass chromatograms obtained from two channel detections that monitor both molecular-related and sphingoid-related ions under two different in-source collision-induced dissociation conditions and using retention times obtained from RPLC. The application of this method revealed that human hair contains 73 species of CER molecules, which were all corroborated by structural analysis using tandem mass spectrometry. The results further revealed that the composition is characterized by predominant molecules consisting of even carbon atom-containing saturated/unsaturated nonhydroxy or a-hydroxy fatty acids and C 18 dihydrosphingosine, a minor but distinct content of isobaric/isomeric and odd chain-containing CERs. This successfully developed RPLC-APCI-MS technique allows the comprehensive profiling of CER molecules in hair for the investigation of their physicochemical and physiological roles.-Masukawa, Y., H. Tsujimura, and H. Narita. Liquid chromatography-mass spectrometry for comprehensive profiling of ceramide molecules in human hair. J. Lipid Res. 2006Res. . 47: 1559Res. -1571 Supplementary key words fatty acid moiety . identification . isobaric .

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“…The optimised LC/ESI/MS method was validated studying several parameters, namely: repeatability, linearity, limit of detection (LOD), limit of quantification (LOQ) and recovery, analysing standard mixtures (10 lM of Cer 6 , Cer 8 , Cer 16 , Cer 18 , Cer 24 : 0 , and Cer 24:1 ) and extracted biological samples supplemented with 10 lM of Cer 2 ceramide as the I.S.…”

Section: Methods Validationmentioning

confidence: 99%

“…Several Cer, namely Cer 2 , Cer 6 , Cer 8 , CER 16 -is probably due to the formation of a dimeric aggregate. The TIC trace also showed a small peak (28.9 min) that presented a mass spectrum resembling that of the studied Cer.…”

Section: Lc-ms Analysismentioning

confidence: 99%

Separation and characterisation of sphingoceramides by high‐performance liquid chromatography‐electrospray ionisation mass spectrometry

Camera

Picardo

Presutti

et al. 2004

J of Separation Science

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We developed a simple and reliable analytical method for the quantification and the characterization of ceramides extracted from biological samples by high-performance liquid chromatography (HPLC) coupled to electrospray ionisation tandem mass spectrometry (ESI/MS/MS). The chromatographic separation of analytes was carried out in a RP8 column, eluting with a methanol-water mixture in gradient elution mode. The separated lipids were detected by total ion monitoring and characterised by MS/MS spectra; quantitative analysis was performed by integrating the extracted ion peaks obtained in the negative ion mode. Good repeatability was obtained for retention time (0.3-2%), peak area ratio (A(S)/A(IS), 2-8%), as well as limit of detection (LOD, 5-26 pg) and quantification (LOQ, 13-53 pg). The method was validated for the analysis of N-palmitoyl-D-erythro-sphingosine (Cer16), N-stearoyl-D-erythro-sphingosine (Cer18), N-tetracosanoyl-D-erythro-sphingosine (N24:0, lignoceric ceramide, Cer24:0), and N-tetracos-15'-enoyl-D-erythro-sphingosine (N24:1, nervonic ceramide, Cer24:1), giving good results. Lipid mixtures, extracted from skin and epidermal cells, were analysed for their content of the studied ceramides.

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Simultaneous quantitative determination method for ceramide species from crude cellular extracts by high-performance liquid chromatography–thermospray mass spectrometry

Cited by 14 publications

References 26 publications

Comprehensive quantification of ceramide species in human stratum corneum

Comprehensive quantification of ceramide species in human stratum corneum

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

Separation and characterisation of sphingoceramides by high‐performance liquid chromatography‐electrospray ionisation mass spectrometry

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