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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

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Lipids

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.

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Abbreviations

d18:2(t4,c8):

trans-4,cis-8-Sphingadienine

d18:2(t4,t8):

trans-4,trans-8-Sphingadienine

ESI:

Electrospray ionization

FID:

Flame ionization detection

GC:

Gas chromatography

Glc:

Glucose

HPLC:

High-performance liquid chromatography

LC:

Liquid chromatography

MS/MS:

Tandem mass spectrometry

ODS:

Octadecylsilyl

t18:1(c8):

4-Hydroxy-cis-8-sphingenine

t18:1(t8):

4-Hydroxy-trans-8-sphingenine

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Acknowledgments

We thank Ayae Wada for technical and experimental assistance. We also thank Dr. Kouhei Yamamoto, Osaka Prefecture University, for his valuable advice. Parts of this work were supported by Special Ordinary Expense Subsidies for Private Universities from the Ministry of Education, Culture, Sports, Science, and Technology (MEXT) of Japan.

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Authors and Affiliations

  1. Department of Biology, Graduate School of Natural Science, Konan University, 8-9-1 Okamoto, Higashinada-ku, Kobe, 658-8501, Japan

    Hiroyuki Imai & Masayuki Watanabe

  2. Development and Marketing Group, Frontier Science Business Division, Shiseido Co., Ltd., 12 Nishikawabe-cho, Higashikujyo, Minami-ku, Kyoto, 601-8037, Japan

    Hideyasu Hattori

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  1. Hiroyuki Imai
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  2. Hideyasu Hattori
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  3. Masayuki Watanabe
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Correspondence to Hiroyuki Imai.

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Imai, H., Hattori, H. & Watanabe, M. An Improved Method for Analysis of Glucosylceramide Species Having cis-8 and trans-8 Isomers of Sphingoid Bases by LC–MS/MS. Lipids 47, 1221–1229 (2012). https://doi.org/10.1007/s11745-012-3725-7

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  • DOI: https://doi.org/10.1007/s11745-012-3725-7

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