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A more recent version of this article appeared on April 1, 2004

Papers In Press, published online ahead of print January 16, 2004
J. Lipid Res., doi:10.1194/jlr.D300036-JLR200
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Submitted on November 11, 2003
Revised on January 6, 2004
Accepted on January 7, 2004

Determination of 24S- and 27-hydroxycholesterol in plasma by high-performance liquid chromatography-mass spectrometry

Ines Burkard, Katharina M. Rentsch, and Arnold von Eckardstein

Institute for Clinical Chemistry, University Hospital Zurich, Zurich, Zurich 8091

Corresponding Author: rentsch{at}ikc.unizh.ch

24S-Hydroxycholesterol (24S-OH-Chol) and 27-hydroxycholesterol (27-OH-Chol) are oxidized derivatives of cholesterol and of potential diagnostic interest since their circulating levels may reflect the cholesterol metabolism of the brain and macrophages, respectively. We developed a sensitive and specific high-performance liquid chromatography-mass spectrometry (HPLC-MS) method for the quantification of 24S-OH-Chol and 27-OH-Chol in human plasma. In contrast to currently available procedures based on gas chromatography-mass spectrometry (GC-MS), this methodology offers the advantage that no time-consuming derivatization is needed. After saponification, solid-phase extraction and HPLC separation under reversed-phase column conditions, detection by MS was performed using atmospheric pressure chemical ionization (APCI) and selected ion monitoring (SIM) mode. The standard curves were linear throughout the calibration range for both oxysterols. Within-day and between-day coefficients of variation were below 9% and the recoveries ranged between 98% and 103%. The respective quantification limits were 40 µg/l and 25 µg/l for 24S-OH-Chol and 27-OH-Chol, respectively. Mean values for both oxysterols were determined in plasma from 22 healthy volunteers. The here described sensitive and selective HPLC-MS method together with the appropriate workup procedure allow the quantification of 24S-OH-Chol and 27-OH-Chol in plasma samples, for example in clinical studies to elaborate the clinical usefulness of these two oxysterols.


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