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Determination of estrogenic compounds in milk and yogurt samples by hollow-fibre liquid-phase microextraction-gas chromatography-triple quadrupole mass spectrometry

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Abstract

An environmentally friendly method based on hollow-fibre liquid-phase microextraction (HF-LPME) was developed for the extraction of selected estrogenic compounds (i.e. four natural sexual hormones: estrone, 17β-estradiol, 17α-estradiol and estriol; two exoestrogens: 17α-ethynylestradiol and 2-methoxyestradiol; two synthetic stilbenes: dienestrol and hexestrol; and five resorcylic acid lactones: zearalenone, α-zearalanol, β-zearalanol, α-zearalenol and β-zearalenol), from whole cow and semi-skimmed goat milk and whole natural yogurt. After the optimization of the sample preparation procedure, spiked extracts were derivatized to their trimethylsilyl products using N,O-bis(trimethylsilyl)trifluoroacetamide reagent and then analyzed by gas chromatography–tandem mass spectrometry (GC-MS/MS). Once optimum extraction conditions were established (protein precipitation with acetonitrile, extraction and the back-extraction in acetonitrile following the HF-LPME procedure), the method was validated and the calibration range, precision and accuracy were studied. The RSD values for the intra- and inter-day precision of the peak areas were in the range 0.65–9.69 and 1.00–11.47 %, respectively. The determination coefficients were higher than 0.991 for method calibration curves while LOD and LOQ values were between 0.06–2.55 and 0.16–6.11 μg/L for whole cow milk, 0.04–1.70 and 0.11–4.86 μg/L for semi-skimmed goat milk and 0.07–3.73 and 0.23–9.81 μg/L for natural yogurt, respectively. Finally, the accuracy and precision of the method were evaluated, obtaining a value in the range 84 81–119 % and RSD values lower than 20 % in all cases.

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Abbreviations

2-MeOE2 :

2-Methoxyestradiol

DES:

Diethylstilbestrol

DS:

Dienestrol

E1 :

Estrone

E2 :

Estradiol

E3 :

Estriol

EE2 :

17α-Ethinylestradiol

HEX:

Hexestrol

ZAL:

Zearalanol

ZEL:

Zearalenol

ZEN:

Zearalenone

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Acknowledgments

This work has been supported by the Spanish Ministry of Economy and Competitiveness (project AGL2011-24667). A.V.H.H. wishes to thank the Fundación CajaCanarias for his grant in the Servicios Generales de Apoyo a la Investigación (SEGAI). G. D’Orazio would like to thank the CNR- Institute of Chemical Methodologies, Monterotondo (Rome, Italy) for the support to his PhD.

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Authors and Affiliations

  1. Institute of Chemical Methodologies, Italian National Research Council-C.N.R., Area della Ricerca di Roma I, Via Salaria Km 29,300, 00015, Monterotondo, Roma, Italy

    Giovanni D’Orazio & Salvatore Fanali

  2. Departamento de Química, Unidad Departamental de Química Analítica, Facultad de Ciencias, Universidad de La Laguna (ULL), Avenida Astrofísico Francisco Sánchez s/n°, 38206, La Laguna, Tenerife, Spain

    Giovanni D’Orazio, Javier Hernández-Borges & Miguel Ángel Rodríguez-Delgado

  3. Instituto Universitario de BioOrgánica Antonio González, Universidad de La Laguna (ULL), Avda. Astrofísico Fco. Sánchez, 2, 38206, La Laguna, Tenerife, Spain

    Antonio Vicente Herrera-Herrera

Authors
  1. Giovanni D’Orazio
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  2. Javier Hernández-Borges
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  3. Antonio Vicente Herrera-Herrera
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  4. Salvatore Fanali
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  5. Miguel Ángel Rodríguez-Delgado
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Correspondence to Miguel Ángel Rodríguez-Delgado.

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D’Orazio, G., Hernández-Borges, J., Herrera-Herrera, A.V. et al. Determination of estrogenic compounds in milk and yogurt samples by hollow-fibre liquid-phase microextraction-gas chromatography-triple quadrupole mass spectrometry. Anal Bioanal Chem 408, 7447–7459 (2016). https://doi.org/10.1007/s00216-016-9833-0

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