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A quantitative HPLC–MS/MS method for studying internal concentrations and toxicokinetics of 34 polar analytes in zebrafish (Danio rerio) embryos

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Abstract

An analytical method using high-performance liquid chromatography–tandem mass spectrometry was developed to determine internal concentrations of 34 test compounds such as pharmaceuticals and pesticides in zebrafish embryos (ZFE), among them, cimetidine, 2,4-dichlorophenoxyacetic acid, metoprolol, atropine and phenytoin. For qualification and quantification, multiple reaction monitoring mode was used. The linear range extends from 0.075 ng/mL for thiacloprid and metazachlor and 7.5 ng/mL for coniine and clofibrate to 250 ng/mL for many of the test compounds. Matrix effects were strongest for nicotine, but never exceeded ±20 % for any of the developmental stages of the ZFE. Method recoveries ranged from 90 to 110 % from an analysis of nine pooled ZFE. These findings together with the simple sample preparation mean this approach is suitable for the determination of internal concentrations from only nine individual ZFE in all life stages up to 96 h post-fertilization. Exemplarily, the time course of the internal concentrations of clofibric acid, metribuzin and benzocaine in ZFE was studied over 96 h, and three different patterns were distinguished, on the basis of the speed and extent of uptake and whether or not a steady state was reached. Decreasing internal concentrations may be due to metabolism in the ZFE.

Quantification of polar pollutants in different life stages of the zebrafish embryo by LC-MS/MS

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Acknowledgments

This work is part of the research topic “Chemicals in the Environment” (CITE) within the research programme of the Helmholtz Centre for Environmental Research–UFZ. We thank C. Petzold and J. Müller for their support in laboratory work and S. Scholz for helpful discussions on ZFE testing and toxicokinetics.

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Authors and Affiliations

  1. Department of Analytical Chemistry, Helmholtz Centre for Environmental Research—UFZ, Permoserstr. 15, 04318, Leipzig, Germany

    Stephan Brox, Axel P. Ritter & Thorsten Reemtsma

  2. Department Bioanalytical Ecotoxicology, Helmholtz Centre for Environmental Research—UFZ, Permoserstr. 15, 04318, Leipzig, Germany

    Eberhard Küster

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  1. Stephan Brox
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  2. Axel P. Ritter
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  3. Eberhard Küster
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  4. Thorsten Reemtsma
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Correspondence to Thorsten Reemtsma.

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Brox, S., Ritter, A.P., Küster, E. et al. A quantitative HPLC–MS/MS method for studying internal concentrations and toxicokinetics of 34 polar analytes in zebrafish (Danio rerio) embryos. Anal Bioanal Chem 406, 4831–4840 (2014). https://doi.org/10.1007/s00216-014-7929-y

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  • DOI: https://doi.org/10.1007/s00216-014-7929-y

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