J. Lipid Res. Neurobiology of Lipids (ISSN1683-5506)
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A more recent version of this article appeared on April 1, 2007

Papers In Press, published online ahead of print January 24, 2007
J. Lipid Res., doi:10.1194/jlr.M600497-JLR200
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Submitted on November 20, 2006
Revised on January 23, 2007
Accepted on January 24, 2007

Liquid chromatography combined with mass spectrometry utilising high-resolution, exact mass, and multi-stage fragmentation for the identification of oxysterols in rat brain

Kersti Karu, Martin Hornshaw, Gary Woffendin, Karl Bodin, Mats Hamberg, Gunvor Alvelius, Jan Sjövall, John Turton, Yuqin Wang, and William J. Griffiths

Pharmaceutical & Biological Chemistry, The School of Pharmacy, London WC1N 1AX

Corresponding Author: william.griffiths{at}pharmacy.ac.uk

In man the brain accounts for about 20% of the body’s free cholesterol, most of which is synthesised de novo in brain. To maintain cholesterol balance throughout life, cholesterol becomes metabolised to 24S-hydroxycholesterol principally in neurons. In mouse, rat, and probably human, metabolism to 24S-hydroxycholesterol accounts for about 50% of cholesterol turnover, however, the route by which the remainder is turned over has yet to be elucidated. Here we describe a novel liquid chromatography (LC) – multi-stage fragmentation mass spectrometry (MSn) methodology for the identification, with high sensitivity (low pg), of cholesterol metabolites in rat brain. The methodology includes derivatisation to enhance ionisation, exact mass analysis at high-resolution to identify potential metabolites, and LC-MS3 to allow their characterisation. 24S-Hydroxycholesterol was confirmed as a major oxysterol in rat brain, while other oxysterols identified for the first time in brain included 24,25-, 24,27-, 25,27-, 6,24, 7alpha,25-, and 7alpha,27-dihydroxycholesterols. In addition, 3beta-hydroxy-5-oxo-5,6-secocholestan-6-al and its aldol, two molecules linked to amyloidogenesis of proteins, were characterised in rat brain.


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W. J. Griffiths, Y. Wang, K. Karu, E. Samuel, S. McDonnell, M. Hornshaw, and C. Shackleton
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[Abstract] [Full Text] [PDF]


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