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Development and validation of a comprehensive two-dimensional gas chromatography–mass spectrometry method for the analysis of phytosterol oxidation products in human plasma

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Abstract

Phytosterol oxidation products (POPs) have been suggested to exert adverse biological effects similar to, although less severe than, their cholesterol counterparts. For that reason, their analysis in human plasma is highly relevant. Comprehensive two-dimensional gas chromatography (GC×GC) coupled with time-of-flight mass spectrometry (TOF-MS) has been proven to be an extremely powerful separation technique for the analysis of very low levels of target compounds in complex mixtures including human plasma. Thus, a GC×GC/TOF-MS method was developed and successfully validated for the simultaneous quantification of ten POPs in human plasma. The calibration curves for each compound showed correlation coefficients (R 2) better than 0.99. The detection limits were below 0.1 ng mL−1. The recovery data were between 71.0% and 98.6% (RSDs <10% for all compounds validated). Good results were obtained for within- and between-day repeatability, with most values being below 10%. In addition, non-targeted sterol metabolites were also identified with the method. The concentrations of POPs found in human plasma in the current study are between 0.3 and 4.5 ng mL−1, i.e., 10–100 times lower than the typical values found for cholesterol oxidation products.

Extracted regions of GC×GC/TOF-MS chromatograms displaying derivatives of sterol oxidation products and sterols for plasma samples

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Authors and Affiliations

  1. Unilever Research and Development, Advanced Measurement and Data Modelling, P.O. Box 114, 3130 AC, Vlaardingen, The Netherlands

    María Menéndez-Carreño, Herrald Steenbergen & Hans-Gerd Janssen

  2. Analytical-Chemistry Group, Van’t Hoff Institute for Molecular Sciences, University of Amsterdam, Science Park 904, P.O. Box 94157, 1090 GD, Amsterdam, The Netherlands

    Hans-Gerd Janssen

Authors
  1. María Menéndez-Carreño
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  2. Herrald Steenbergen
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  3. Hans-Gerd Janssen
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Correspondence to María Menéndez-Carreño.

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Published in the special issue Comprehensive Multidimensional Separations with Guest Editors James Harynuk and Philip Marriott.

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Menéndez-Carreño, M., Steenbergen, H. & Janssen, HG. Development and validation of a comprehensive two-dimensional gas chromatography–mass spectrometry method for the analysis of phytosterol oxidation products in human plasma. Anal Bioanal Chem 402, 2023–2032 (2012). https://doi.org/10.1007/s00216-011-5432-2

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  • DOI: https://doi.org/10.1007/s00216-011-5432-2

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