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A more recent version of this article appeared on July 1, 2006

Papers In Press, published online ahead of print April 25, 2006
J. Lipid Res., doi:10.1194/jlr.D600007-JLR200
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Submitted on February 14, 2006
Revised on April 18, 2006
Accepted on April 24, 2006

Reversed phase liquid chromatography atmospheric pressure chemical ionization mass spectrometry for comprehensive profiling of ceramide molecules in human hair

Yoshinori Masukawa, Hisashi Tsujimura, and Hirofumi Narita

Kao Analytical Research Center, Kao Corporation, Haga-Gun, Tochigi 321-3497

Corresponding Author: masukawa.yoshinori{at}kao.co.jp

Ceramides (CERs), which consist of various fatty acid and sphingoid moieties, 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 mixed CER molecules including isobaric and isomeric CERs. Therefore, a method for the comprehensive profiling of CERs was developed and then applied to the analysis of CERs in the hair. The method developed is based on reversed phase liquid chromatography (RPLC) coupled to atmospheric pressure chemical ionization (APCI) MS with positive ion mode. Comprehensive identification and profiling of CERs is achieved by using of two sets of multi-mass chromatograms obtained from two channel detections that monitor both molecular-related and sphingoid-related ions under two different in-source collision-induced dissociation conditions upon the positive APCI, and by using retention times obtained from RPLC. The application of this method to the analysis of CERs in the hair revealed that human hair contains 73 species of CER molecules, which were all corroborated by structural analysis using negative ESI tandem mass spectrometry. The results further revealed that the composition is characterized by predominant CER molecules consisting of even carbon atom-containing saturated/unsaturated non-hydroxy or a-hydroxy fatty acids and C18 dihydrosphingosine, a minor but distinct content of isobaric/isomeric and odd chain-containing CERs, in which branched chain-containing C17 and C19 dihydrosphingosine might be included. This successfully developed RPLC-APCI-MS technique allows the comprehensive profiling of mixed CER molecules in the hair for the investigation of their physicochemical and physiological roles.


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