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Focused ultrasound-assisted acceleration of enzymatic hydrolysis of alkylphenols and 17β-oestradiol glucuronide in fish bile

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Abstract

According to the European Water Framework Directive (WFD), alkylphenols, such as octylphenols and nonylphenols, and 17β-oestradiol are considered as priority or emerging pollutants, respectively, mainly due to their possible properties as endocrine-disrupting compounds (EDCs). EDCs are accumulated in liver, fat, kidney and bile in the glucuronide form. In order to determine the concentration of these compounds in bile, an enzymatic hydrolysis step is necessary. This step is usually long (∼16 h), and in this sense, ultrasound probes were studied as a possible alternative energy source to accelerate this process. Enzymatic hydrolysis was reduced to 20 min using an ultrasound probe at one cycle and 10% of amplitude. For validation of analytical procedure, nonylphenol glucuronide (4NP-G), 4-tert-octylphenol glucuronide (4tOP-G) and 4-n-octylphenol glucuronide (4nOP-G) were synthesised while 17β-oestradiol glucuronide (E2-G) was commercially available. Bile from thick-lip grey mullets (Chelon labrosus) was spiked with known amounts of 4NP-G, 4tOP-G, 4nOP-G and E2-G and submitted to the optimised procedure. Good recoveries (77–122%), precision in the 5% to 12% range and limits of detection, ranging from the low nanogramme per gramme level for 4tOP, 4nOP and E2 to the low microgramme per gramme level for nonylphenols, were obtained. The optimised method was applied for the determination of alkylphenol in the bile of thick-lip grey mullets fish bile from the Urdaibai estuary (UNESCO reserve of the Biosphere, Bay of Biscay), and high concentrations (2.3–14.2 μg/g), such as those obtained in polluted areas, were measured. E2 was determined in the bile of thick-lip grey mullets, intraperitoneally injected with E2.

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Acknowledgements

This work was supported by the Spanish Ministry of Science through the CTQ2008-02775/BQU project and the University of the Basque Country through the UNESCO 07/09 project. A. Vallejo is grateful to the University of the Basque Country for his pre-doctoral fellowship.

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Authors and Affiliations

  1. Department of Analytical Chemistry, University of Basque Country, 644 PK, 48080, Bilbao, Spain

    Asier Vallejo, Aresatz Usobiaga, Luis A. Fernández & Olatz Zuloaga

  2. Department of Zoology and Animal Cell Biology, University of the Basque Country, 48080, Bilbao, Spain

    Maren Ortiz-Zarragoitia & Miren P. Cajaraville

Authors
  1. Asier Vallejo
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  2. Aresatz Usobiaga
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  3. Maren Ortiz-Zarragoitia
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  4. Miren P. Cajaraville
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  5. Luis A. Fernández
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  6. Olatz Zuloaga
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Correspondence to Asier Vallejo.

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Vallejo, A., Usobiaga, A., Ortiz-Zarragoitia, M. et al. Focused ultrasound-assisted acceleration of enzymatic hydrolysis of alkylphenols and 17β-oestradiol glucuronide in fish bile. Anal Bioanal Chem 398, 2307–2314 (2010). https://doi.org/10.1007/s00216-010-4156-z

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  • DOI: https://doi.org/10.1007/s00216-010-4156-z

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