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Method for quantifying NSAIDs and clofibric acid in aqueous samples, lumpfish (Cyclopterus lumpus) roe, and zebrafish (Danio rerio) eleutheroembryos and evaluation of their bioconcentration in zebrafish eleutheroembryos

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Abstract

Pharmaceuticals such as nonsteroidal anti-inflammatory drugs (NSAIDs) and lipid regulators are being repeatedly detected at low concentrations (pg · mL−1–ng · mL−1) in the environment. A large fraction of these compounds are ionizable. Ionized compounds show different physico-chemical properties and environmental behavior in comparison to their neutral analogs; as a consequence, the quantification methods currently available, based on the neutral molecules, might not be suitable to detect the corresponding ionized compounds. To overcome this problem, we developed a specific analytical method to quantify NSAIDs and lipid regulators (i.e., ibuprofen, diclofenac, naproxen, and clofibric acid) and their ionized compounds. This method is based on three steps: (1) the extraction of the organic compounds with an organic solvent assisted with an ultrasonic probe, (2) the cleaning of the extracts with a dispersive SPE with C18, and (3) the determination of the chemical compounds by GC-MS (prior derivatization of the analytes). We demonstrated that the proposed method can successfully quantify the pharmaceuticals and their ionized compounds in aqueous samples, lumpfish eggs, and zebrafish eleutheroembryos. Additionally, it allows the extraction and the cleanup of extracts from small samples (0.010 g of wet weight in pools of 20 larvae) and complex matrixes (due to high lipid content) and can be used as a basis for bioaccumulation assays performed with zebrafish eleutheroembryos in alternative to OECD test 305.

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Acknowledgments

The authors want to thank the financial support of this work by the Ministry of Economy and Competitivity (Project SAFEFOOD, grant code CTQ2014-54801-C2-1-R), Comunidad Autónoma of Madrid (Project AVANSECAL, code S2013/ABI-3028) and Universidad Complutense for supporting the research group “Determinación de Trazas y Especiación.” Noemí Molina thanks the Spanish Ministry of Education for the FPU predoctoral fellowship (Ref. AP2010-0740). We are grateful to AZTI (Bizkaia, Spain) for providing the eleutheroembryo samples.

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  1. Department of Analytical Chemistry, Faculty of Chemical Science, Complutense University of Madrid, Avenida Complutense s/n, 28040, Madrid, Spain

    N. Molina-Fernandez, C. Perez-Conde & J. Sanz-Landaluze

  2. Food Research Division, AZTI, Parque Tecnológico de Bizkaia, Astondo Bidea 609, 48160, Derio, Spain

    S. Rainieri

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  1. N. Molina-Fernandez
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  2. C. Perez-Conde
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  4. J. Sanz-Landaluze
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Correspondence to J. Sanz-Landaluze.

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ESM 1

Fig. S1 Analyte areas obtained using different derivatization time for a solution of four compounds (CFB, IBU, NP, DC) at 1 μg · mL−1. Table S1 Recoveries and %RSD obtained for the four pharmaceuticals in an intra and inter-day study at two different concentrations for the three different matrices. (DOC 88 kb)

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Molina-Fernandez, N., Perez-Conde, C., Rainieri, S. et al. Method for quantifying NSAIDs and clofibric acid in aqueous samples, lumpfish (Cyclopterus lumpus) roe, and zebrafish (Danio rerio) eleutheroembryos and evaluation of their bioconcentration in zebrafish eleutheroembryos. Environ Sci Pollut Res 24, 10907–10918 (2017). https://doi.org/10.1007/s11356-016-6671-8

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