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Separation of Δ5- and Δ7-Phytosterols by Adsorption Chromatography and Semipreparative Reversed Phase High-Performance Liquid Chromatography for Quantitative Analysis of Phytosterols in Foods

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    Separation of Δ5- and Δ7-Phytosterols by Adsorption Chromatography and Semipreparative Reversed Phase High-Performance Liquid Chromatography for Quantitative Analysis of Phytosterols in Foods
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    Laboratoire de Chimie Analytique et Sciences de l'Aliment (LC4, UMR 7178), Faculté de Pharmacie, Université Louis Pasteur, 74 route du Rhin, 67400 Illkirch, France, Laboratoire de Chimie Organique Synthétique (LC3, UMR 7177), Université Louis Pasteur, 1 rue Blaise Pascal, 67008 Strasbourg, France, Laboratoire d'Oncologie Nutritionnelle, ULP-EA 3430, IRCAD, 1 place de l'Hôpital, 67091 Strasbourg, France, and Aérial, rue Laurent Fries, Parc d'Innovation, 67412 Illkirch, France
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    Journal of Agricultural and Food Chemistry

    Cite this: J. Agric. Food Chem. 2006, 54, 4, 1196–1202
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    https://doi.org/10.1021/jf052761x
    Published January 31, 2006
    Copyright © 2006 American Chemical Society
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    Abstract

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    A method for the separation, isolation, and identification of phytosterols was developed. A commercial phytosterols mixture, Generol 95S, was fractionated first by adsorption silica gel column chromatography and then separated by means of a semipreparative reverse phase high-performance liquid chromatography fitted with a Polaris C8-A column (250 mm × 10 mm i.d., 5 μm) using isocratic acetonitrile:2-propanol:water (2:1:1, v/v/v) as the mobile phase. Milligram scales of six individual phytosterols, including citrostadienol, campesterol, β-sitosterol, Δ7-avenasterol, Δ7-campesterol, and Δ7-sitosterol, were obtained. Purities of these isolated sterols were 85−98%. Relative response factors (RRF) of these phytosterols were calculated against cholestanol as an authentic commercial standard. These RRF values were used to quantify by gas chromatography−mass spectrometry (GC-MS) the phytosterols content in a reference material, oils, and chocolates.

    Keywords: Phytosterol; isolation; column chromatography; semipreparative RP-HPLC; quantitative analysis

    Copyright © 2006 American Chemical Society

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    †

     Laboratoire de Chimie Analytique et Sciences de l'Aliment (LC4, UMR 7178), Université Louis Pasteur.

    ‡

     Laboratoire de Chimie Organique Synthétique (LC3, UMR 7177), Université Louis Pasteur.

    §

     IRCAD.

    ‖

     Aérial.

    *

     To whom correspondence should be addressed. Tel:  +33(0)3 90244326. Fax:  +33(0)3 90244325. E-mail:  [email protected].

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    Journal of Agricultural and Food Chemistry

    Cite this: J. Agric. Food Chem. 2006, 54, 4, 1196–1202
    Click to copy citationCitation copied!
    https://doi.org/10.1021/jf052761x
    Published January 31, 2006
    Copyright © 2006 American Chemical Society
    Request reuse permissions

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