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A more recent version of this article appeared on July 1, 2003

Papers In Press, published online ahead of print May 1, 2003
J. Lipid Res., doi:10.1194/jlr.D300010-JLR200
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Submitted on March 27, 2003
Revised on April 28, 2003
Accepted on April 29, 2003

Simultaneous quantification of lyso-neutral-glycosphingolipids and neutral glycosphingolipids by N-acetylation with [3H]acetic anhydride

Jacques Bodennec, Selena Trajkovic-Bodennec, and Anthony H. Futerman

Biol. Chem., Weizmann Institute, Rehovot 76-100

Corresponding Author: tony.futerman{at}weizmann.ac.il

We describe a new method that permits quantification in the pmol to nmol range of three lyso-neutral-glycosphingolipids (lyso-n-GSLs), glucosylsphingosine (GlcSph), galactosylsphingosine (GalSph) and lactosylsphingosine (LacSph), in the same sample as neutral-glycosphingolipids (n-GSLs). Lyso-n-GSLs and n-GSLs are initially obtained from a crude lipid extract using Sephadex G25 chromatography, followed by their isolation in one fraction, which is devoid of other contaminating lipids, by aminopropyl solid phase chromatography. Lyso-n-GSLs and n-GSLs are subsequently separated from one another by weak cation exchange chromatography. n-GSLs are then deacylated by strong alkaline hydrolysis, and the N-deacylated-GSLs and lyso-n-GSLs are subsequently N-acetylated using [3H]acetic anhydride. An optimal concentration of 5 mM acetic anhydride was established, which gave >95% N-acetylation. We demonstrate the usefulness of this technique by showing an ~40-fold increase of both GlcSph and glucosylceramide in brain tissue from a glucocerebrosidase-deficient mouse, as well as significant lactosylceramide accumulation. The application and optimization of this technique for lyso-n-GSLs and lyso-GSLs will permit their quantification in small amounts of biological tissues, particularly in the GSL storage diseases, such as Gaucher and Krabbe’s disease, in which GlcSph and GalSph, respectively, accumulate.


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This article has been cited by other articles:


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[Abstract] [Full Text] [PDF]


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