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A more recent version of this article appeared on January 1, 2005

Papers In Press, published online ahead of print October 16, 2004
J. Lipid Res., doi:10.1194/jlr.D400022-JLR200
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Submitted on August 23, 2004
Revised on October 12, 2004
Accepted on October 14, 2004

Characterization and direct quantitation of cerebroside molecular species from lipid extracts by shotgun lipidomics

Xianlin Han and Hua Cheng

Dept of Internal Medicine, Washington University School of Medicine, St. Louis, MO 63110

Corresponding Author: xianlin{at}wustl.edu

By employing shotgun lipidomics based on the separation of lipid classes in the electrospray ion source (intrasource separation) and 2D mass spectrometry (MS) techniques (Mass Spectrom. Rev. DOI: 10.1002/mas.20023), individual molecular species of most major and many minor lipid classes can be quantitated directly from biological lipid extracts. Herein, we extended shotgun lipidomics to the characterization and quantitation of cerebroside molecular species in biological samples. By exploiting the differential fragmentation patterns of chlorine adducts using electrospray ionization (ESI) tandem mass spectrometry, hydroxy and non-hydroxy cerebroside species are readily identified. The hexose (either galactose or glucose) moiety of a cerebroside species can be distinguished by examination of the peak intensity ratio of its product ions at m/z 179 and 89 (i.e., 0.74 ± 0.10 and 4.8 ± 0.7 for galactose- and glucose-containing cerebroside species, respectively). Quantitation of cerebroside molecular species (as little as 10 fmol) from chloroform extracts of brain tissue samples was directly conducted by 2D ESI/MS after correction for differences in 13C isotopomer intensities. This method was demonstrated to have over a 1000-fold linear dynamic range in the low concentration region, and therefore, it should have a wide range of applications in studies on the cellular sphingolipid lipidome.


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