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Anal Biochem. 2008 Oct 1;381(1):151-3. doi: 10.1016/j.ab.2008.05.037. Epub 2008 Jun 13.

Liquid chromatography-tandem mass spectrometry determination of plasma 24S-hydroxycholesterol with chromatographic separation of 25-hydroxycholesterol.

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  • 1Department of Physiology and Pharmacology, Oregon Health and Science University, Portland, OR 97239, USA.


Concentrations of circulating 24S-hydroxycholesterol (24SOHChol) are of interest as a practical measure of cholesterol efflux from the human brain. The current method of choice for 24SOHChol quantification is with gas chromatography-mass spectrometry (GC-MS). Liquid chromatography-mass spectrometry (LC-MS) methods to detect 24SOHChol have been described, but they lack rigorous high-performance liquid chromatography (HPLC) separation of a closely eluting isomeric oxysterol, 25-hydroxycholesterol. This is important because 25-hydroxycholesterol can be present in significant amounts and tandem mass spectrometry (MS/MS) cannot completely differentiate 24SOHChol. We describe an LC-MS method with rapid chromatographic separation of the oxysterols to permit accurate determination of plasma 24SOHChol. The availability of an LC-MS method offers advantages such as simplified sample work-up and analysis.

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