Liquid Chromatography-mass Spectrometry Utilizing Multi-stage Fragmentation for the Identification of Oxysterols

Overview

Journal J Lipid Res

Publisher Elsevier

Specialty Biochemistry

Date 2007 Jan 26

PMID 17251593

Citations 33

Authors

Kersti Karu

Martin Hornshaw

Gary Woffendin

Karl Bodin

Mats Hamberg

Gunvor Alvelius

Jan Sjovall

John Turton

Yuqin Wang

William J Griffiths

Affiliations

Soon will be listed here.

Abstract

In humans, the brain accounts for about 20% of the body's free cholesterol, most of which is synthesized de novo in brain. To maintain cholesterol balance throughout life, cholesterol becomes metabolized 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 (MS(n)) methodology for the identification, with high sensitivity (low pg), of cholesterol metabolites in rat brain. The methodology includes derivatization to enhance ionization, exact mass analysis at high resolution to identify potential metabolites, and LC-MS(n) (n=3) to allow their characterization. 24S-hydroxycholesterol was confirmed as a major oxysterol in rat brain, and 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 characterized in rat brain.

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