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Journal of Lipid Research, Vol. 46, 21-26, January 2005
Copyright © 2005 by American Society for Biochemistry and Molecular Biology

Rapid Communication

Rapid quantification of free and esterified phytosterols in human serum using APPI-LC-MS/MS

Jan Lembcke¹, Uta Ceglarek¹, Georg Martin Fiedler, Sven Baumann, Alexander Leichtle and Joachim Thiery²

Institute of Laboratory Medicine, Clinical Chemistry, and Molecular Diagnostics, University Hospital, Leipzig, Germany

² To whom correspondence should be addressed. e-mail: thiery{at}medizin.uni-leipzig.de

ABSTRACT

A novel analytical platform based on liquid chromatography and tandem mass spectrometry using atmospheric pressure photoionization 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 3 h [gas chromatography-mass spectrometry (GC-MS)] to 15 min. The detection limits of the different phytosterols ranged between 0.25 and 0.68 µg/l. Linear ranges were between 1 and 1,000 µg/l. The within-run and between-run variabilities ranged between 1.4% and 9.9%. The analytical sensitivity was at least 150-fold higher compared with GC-MS.

Our new method allows a rapid and simultaneous determination of free and esterified phytosterols in serum.

Abbreviations: APPI, atmospheric pressure photoionization; LC-MS/MS, liquid chromatography-tandem mass spectrometry; LPDS, lipoprotein-deficient serum

Supplementary key words free phytosterols • phytosterol esters • liquid chromatography • plant sterols

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