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An SPME–GC–MS method using an octadecyl silica fibre for the determination of the potential angiogenesis modulators 17β-estradiol and 2-methoxyestradiol in culture media

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Abstract

A simple and easily automable method based on solid-phase microextraction followed by gas chromatographic–mass spectrometric analysis was developed for the determination of two potential angiogenesis modulators 17β-estradiol (17-BE) and 2-methoxyestradiol (2-MEOE) in culture media. Trifluoroacetic anhydride was used as the derivatising agent. A homemade octadecyl silica coating, characterised by a coating thickness of 72 ± 10 μm and a good thermal stability until 250 °C, was prepared. Experimental design was used to optimise the extraction conditions in terms of derivatisation time, derivatisation temperature and time of extraction. As for method validation, lower limits of quantification of 0.17 and 0.015 µg/l for 17β-estradiol and 2-methoxyestradiol, respectively, were obtained. Finally, the capabilities of the developed fibres were evaluated for the analysis of the investigated analytes developed by granulosa cells in culture media maintained under normoxic, hypoxic and anoxic conditions, in order to better elucidate their possible role in the angiogenic process. An increase of the production of both 17-BE and 2-MEOE in hypoxic and anoxic conditions seems to be related to the effect of oxygen deprivation.

17-βestradiol and 2-methoxyestradiol as potential angiogenesis modulators

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Authors and Affiliations

  1. Dipartimento di Chimica Generale ed Inorganica, Chimica Analitica, Chimica Fisica, Università degli Studi di Parma, Viale Usberti 17/A, 43100, Parma, Italy

    Federica Bianchi, Monica Mattarozzi, Maria Careri, Alessandro Mangia & Marilena Musci

  2. Dipartimento di Produzioni Animali, Biotecnologie Veterinarie, Qualità e Sicurezza degli Alimenti, Sezione di Fisiologia Veterinaria, Università degli Studi di Parma, Via del Taglio 8, 43100, Parma, Italy

    Francesca Grasselli, Simona Bussolati & Giuseppina Basini

Authors
  1. Federica Bianchi
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  2. Monica Mattarozzi
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  3. Maria Careri
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  4. Alessandro Mangia
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  5. Marilena Musci
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  6. Francesca Grasselli
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  7. Simona Bussolati
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  8. Giuseppina Basini
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Correspondence to Federica Bianchi.

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Bianchi, F., Mattarozzi, M., Careri, M. et al. An SPME–GC–MS method using an octadecyl silica fibre for the determination of the potential angiogenesis modulators 17β-estradiol and 2-methoxyestradiol in culture media. Anal Bioanal Chem 396, 2639–2645 (2010). https://doi.org/10.1007/s00216-010-3508-z

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  • DOI: https://doi.org/10.1007/s00216-010-3508-z

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