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Papers In Press, published online ahead of print December 1, 2008
J. Lipid Res., doi:10.1194/jlr.D800038-JLR200

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Journal of Lipid Research, Vol. 49, 2678-2689, December 2008
Copyright © 2008 by American Society for Biochemistry and Molecular Biology

Methods

Targeted analysis of ganglioside and sulfatide molecular species by LC/ESI-MS/MS with theoretically expanded multiple reaction monitoring

Kazutaka Ikeda^*,, Takao Shimizu and Ryo Taguchi^1,*,

^* Department of Metabolome, Graduate School of Medicine, Faculty of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan
Department of Biochemistry and Molecular Biology, Faculty of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan
Core Research for Evolutional Science and Technology, Japan Science and Technology Agency, Kawaguchi, Saitama 332-0012, Japan

The online version of this article (available at http://www.jlr.org) contains supplementary data in the form of one figure.

Published, JLR Papers in Press, August 14, 2008.

This study was performed with the help of Core Research for Evolutional Science and Technology, Japan Science and Technology Agency.

¹ To whom correspondence should be addressed. e-mail: rytagu{at}m.u-tokyo.ac.jp

Matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) has been applied primarily to the analysis of glycosphingolipids separated from other complex mixtures by TLC, but it is difficult to obtain quantitative profiling of each glycosphingolipid among the different spots on TLC by MALDI-MS. Thus, the development of a convenient approach that utilizes liquid chromatography/electrospray ionization (LC/ESI)-MS has received interest. However, previously reported methods have been insufficient to separate and distinguish each ganglioside class. Here we report an effective method for the targeted analysis of theoretically expected ganglioside molecular species by LC/ESI tandem mass spectrometry (LC/ESI-MS/MS) in combination with multiple reaction monitoring (MRM). MRM detection specific for sialic acid enabled us to analyze ganglioside standards such as GM1, GM2, GM3, GD1, and GT1 at picomolar to femtomolar levels. Furthermore, other gangliosides, such as GD2, GD3, GT2, GT3, and GQ1, were also detected in glycosphingolipid standard mixtures from porcine brain and acidic glycolipid extract from mouse brain by theoretically expanded MRM. We found that this approach was also applicable to sulfatides contained in the glycosphingolipid mixtures. In addition, we established a method to separate and distinguish regioisomeric gangliosides, such as GM1a and -1b, GD1a, -1b, and -1c, and GT1a, -1b, and -1c with diagnostic sugar chains in the MRM.

Supplementary key words liquid chromatography/electrospray ionization tandem mass spectrometry • glycosphingolipids • gangliosides • sulfatides • lipidomics

Abbreviations: CID, collision-induced dissociation; DP, declustering potential; ESI, electrospray ionization; LC, liquid chromatography; LCB, long-chain base; MRM, multiple reaction monitoring; MS, mass spectrometry; MS/MS, tandem mass spectrometry; ODS, octadecyl silane; RPLC, reverse-phase LC

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