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An Improved Method for Analysis of Glucosylceramide Species Having cis ‐8 and trans ‐8 Isomers of Sphingoid Bases by LC–MS/MS
Author(s) -
Imai Hiroyuki,
Hattori Hideyasu,
Watanabe Masayuki
Publication year - 2012
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/s11745-012-3725-7
Subject(s) - chromatography , chemistry , electrospray ionization , high performance liquid chromatography , mass spectrometry , tandem mass spectrometry , lipidomics , gas chromatography , biochemistry
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.