Methods

Lipids

, Volume 47, Issue 12, pp 1221-1229

An Improved Method for Analysis of Glucosylceramide Species Having cis-8 and trans-8 Isomers of Sphingoid Bases by LC–MS/MS

  • Hiroyuki ImaiAffiliated withDepartment of Biology, Graduate School of Natural Science, Konan University Email author 
  • , Hideyasu HattoriAffiliated withDevelopment and Marketing Group, Frontier Science Business Division, Shiseido Co., Ltd.
  • , Masayuki WatanabeAffiliated withDepartment of Biology, Graduate School of Natural Science, Konan University

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Abstract

High-performance liquid chromatography–electrospray ionization tandem mass spectrometry (HPLC–ESI–MS/MS) approaches have enabled high selectivity and sensitivity for the identification and quantification of glucosylceramide molecular species. Here we demonstrate that HPLC–ESI–MS/MS is an efficient method for characterizing plant glucosylceramide species having the cis-8 and trans-8 isomers of sphingoid bases. Complete baseline separation was achieved using a high-carbon-content octadecylsilyl column and a simple binary gradient comprising methanol and water. The result of 2-hydroxy fatty acid composition achieved by HPLC–ESI–MS/MS was compared with that achieved by gas chromatography with flame ionization detection (GC–FID), indicating that the two methods yield similar molar compositions. The current method should be applicable to seeking the active components of glucosylceramide species from plant materials in response to biological challenges.

Keywords

Glucosylceramides Glycosphingolipids Liquid chromatography–electrospray ionization tandem mass spectrometry Plant lipids Sphingoid bases