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A more recent version of this article appeared on January 1, 2005

Papers In Press, published online ahead of print October 16, 2004
J. Lipid Res., doi:10.1194/jlr.C400004-JLR200
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Submitted on June 28, 2004
Revised on September 27, 2004
Accepted on October 14, 2004

Rapid quantification of free and esterified phytosterols in serum using atmospheric pressure photoionization liquid chromatography tandem mass spectrometry

Jan Lembcke, Uta Ceglarek, Georg Martin Fiedler, Sven Baumann, Alexander Leichtle, and Joachim Thiery

Department of Clinical Chemistry and Molecular Diagnostics, University Hospital Leipzig, Leipzig D-04103

Corresponding Author: thiery{at}medizin.uni-leipzig.de

Recent studies suggest that even modest elevations of serum phytosterols may be associated with an increased atherosclerotic risk. However, further large-scale studies are mandatory to prove the role of phytosterols as an additional atherosclerotic risk factor. For this purpose sensitive high throughput analytical methods are necessary. A novel analytical platform based on liquid chromatography and tandem mass spectrometry (LC-MS/MS) using atmospheric pressure photoionization (APPI) was applied for the simultaneous quantification of free and esterified ß-sitosterol, campesterol, brassicasterol, and stigmasterol. The total time for sample pretreatment and analysis could be reduced from about three hours (gas chromatography-mass spectrometry, GC-MS) to 15 minutes. The detection limits of the different phytosterols ranged between 0.25 and 0.68 µg/l. Linear ranges were between 1 and 1000 µg/l. The within and between-run variability ranged between 1.4 and 9.9%. The analytical sensitivity was at least 150fold higher compared to GC-MS. Our new method allows a rapid and simultaneous determination of free and esterified phytosterols in serum.


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