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Distribution of C16:0, C18:0, C24:1, and C24:0 sulfatides in central nervous system lipid rafts by quantitative ultra-high-pressure liquid chromatography tandem mass spectrometry

Journal article
Authors A. L. Moyano
G. N. Li
A. Lopez-Rosas
Jan-Eric Månsson
R. B. van Breemen
M. I. Givogri
Published in Analytical Biochemistry
Volume 467
Pages 31-39
ISSN 0003-2697
Publication year 2014
Published at Institute of Neuroscience and Physiology, Department of Psychiatry and Neurochemistry
Pages 31-39
Language en
Keywords Sulfated galactosylceramides, Membrane microdomains, Tandem mass spectrometry, Metachromatic, A-DEFICIENT MICE, METACHROMATIC LEUKODYSTROPHY, MULTIPLE-SCLEROSIS, CELL-MEMBRANES, DETERGENT, BRAIN, SULFOGLYCOLIPIDS, DOMAINS, DISEASE, TISSUE, Biochemical Research Methods, Biochemistry & Molecular Biology, Chemistry, Analytical
Subject categories Clinical Medicine


Sulfated galactosylceramides (sulfatides) are glycosphingolipids associated with cholesterol- and sphingolipid-enriched membrane microdomains (lipid rafts) and are highly expressed in brain tissue. Although it is known that sulfatide species show heterogeneity in their fatty acid acyl group composition throughout brain development, their lipid raft distribution and biological relevance is poorly understood. We validated a fast and sensitive ultra-high-pressure liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method to measure developmentally regulated sulfatide species (06:0, C18:0, C24:1, and C24:0) in central nervous system (CNS) lipid rafts isolated without using detergent. Our UHPLC-MS/MS assay showed good accuracy and precision with a linear range of 5 to 1000 nM for C18:0 and C24:1 sulfatides and 10 to 1000 nM for 06:0 and C24:0 sulfatides. We applied this quantitative analysis to detergent-free lipid rafts isolated from wild-type mice and arylsulfatase A-deficient (ASA knockout) mice that accumulate sulfatides. All four sulfatide species were more abundant in raft membranes than in non-raft membranes, with a significant increase in lipid rafts isolated from ASA knockout mice. This is the first description of an analytical method to study these sulfatide species in raft and non-raft membranes and has the potential to be applied to preparations from other tissues.

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