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Determination of Fecal Sterols Following a Diet with and without Plant Sterols

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Lipids

Abstract

The aim of this study was to develop a method for neutral fecal sterols determination in subjects receiving a normal diet with or without a plant sterols-enriched beverage using gas chromatography–mass spectrometry (GC/MS). Sample preparation conditions (homogenization of lyophilized feces with water) were evaluated. Sterol determination required direct hot saponification, unsaponifiable extraction with hexane, and the formation of trimethylsilyl (TMS) ether derivatives. The method allows the quantification of cholesterol, plant sterols and their metabolites (coprostanol, coprostanone, cholestanol, cholestanone, methylcoprostanol, methylcoprostanone, ethylcoprostenol, stigmastenol, ethylcoprostanol and ethylcoprostanone). Good linearity was obtained (r > 0.96) and interference was only observed for coprostanone, where the standard addition method proved necessary for quantification. The limits of detection (LOD) ranged from 0.10 to 3.88 µg/g dry feces and the limits of quantitation (LOQ) from 0.34 to 12.94 µg/g dry feces. Intra- and inter-assay precision (RSD %) were 0.9–9.2 and 2.1–11.3, respectively. Accuracy, expressed as percentage recovery (80–119%) was obtained for all determined sterols.

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Abbreviations

PS:

Plant sterols

GC/MS:

Gas chromatography–mass spectrometry

RSD:

Relative standard deviation

LOD:

Limit of detection

LOQ:

Limit of quantitation

IS:

Internal standard

BHT:

Butylhydroxytoluene

BSTFA:

N,O-Bis(trimethylsilyl)trifluoroacetamide

TMCS:

Trimethylchlorosilane

TMS:

Trimethylsilyl

Rrt:

Relative retention time

Rt:

Retention time

NIST:

National Institute of Standards and Technology

SD:

Standard deviation

AOAC:

Association of Official Analytical Chemists

Brassicasterol:

5,22-Cholestadien-24β-methyl-3β-ol

Campestanol:

24α-Methyl-5β-cholestan-3β-ol

Campesterol:

24α-Methyl-5-cholesten-3β-ol

Coprostanol:

5β-Cholestan-3β-ol

Coprostanone:

5β-Cholestan-3-one

Cholestane:

5α-Cholestane

Cholestanol:

5α-Cholestan-3β-ol

Cholestanone:

5α-Cholestan-3-one

Cholestenone:

4-Cholesten-3-one

Cholesterol:

5-Cholesten-3β-ol

Cholesterone:

5-Cholesten-3-one

Desmosterol:

5,24-Cholestadien-3β-ol

Epicoprostanol:

5β-Cholestan-3α-ol

Ethylcoprostanol:

24β-Ethyl-5β-cholestan-3β-ol

Ethylcoprostanone:

24β-Ethyl-5β-cholestan-3-one

Ethylcoprostenol:

24β-Ethyl-5β-cholestan-3α-ol

Lathosterol:

5α-Cholest-7-en-3β-ol

β-Sitosterol:

5-Stigmansten-3β-ol

Methylcoprostanol:

24α-Methyl-5β-cholestan-3β-ol

Methylcoprostanone:

24α-Methyl-5β-cholestan-3-one

Stigmastanol:

3β-5α-Stigmastan-3-ol

Stigmastenol:

5α-Stigmast-7-en-3β-ol

Stigmasterol:

Stigmastan-5,22-dien-3β-ol

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Acknowledgements

Thanks are due to the Vitamins Unit of Puerta del Hierro-Majadahonda University Hospital (Madrid, Spain) for providing the feces samples, and to Hero España, S.A., for manufacturing the beverages. This study was part of a project funded by the Spanish Ministry of Economy and Competitiveness (AGL2012-39503-C02-01)(MINECO-FEDER). Maria Cuevas-Tena holds a grant from the Spanish Ministry of Economy and Competitiveness (BES-2013-062705) MINECO.

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Authors and Affiliations

  1. Nutrition and Food Chemistry Area, Faculty of Pharmacy, University of Valencia, Avda. Vicente Andrés Estellés s/n, Burjassot, Valencia, 46100, Spain

    María Cuevas-Tena, Amparo Alegría & María Jesús Lagarda

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  1. María Cuevas-Tena
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  2. Amparo Alegría
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  3. María Jesús Lagarda
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Correspondence to Amparo Alegría.

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Cuevas-Tena, M., Alegría, A. & Lagarda, M.J. Determination of Fecal Sterols Following a Diet with and without Plant Sterols. Lipids 52, 871–884 (2017). https://doi.org/10.1007/s11745-017-4286-6

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