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J. Lipid Res.
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A more recent version of this article appeared on December 1, 2008

Papers In Press, published online ahead of print August 14, 2008
J. Lipid Res., doi:10.1194/jlr.D800038-JLR200
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Submitted on July 22, 2008
Revised on August 5, 2008
Accepted on August 14, 2008

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

Department of Metabolome, Graduate School of Medicine, The University of Tokyo, Tokyo

Corresponding Author: rytagu{at}m.u-tokyo.ac.jp

Matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) was mostly applied to the analysis of glycosphingolipids separated from the other complex mixtures by thin-layer chromatography (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, but previously reported methods were 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-MS/MS combination with multiple reaction monitoring (MRM). MRM detection specific for sialic acid enables 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, 1b, GD1a, 1b, 1c and GT1a, 1b, 1c with diagnostic sugar chains in the MRM.


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