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Optimization and comparison of miniaturized extraction techniques for PAHs from crude oil exposed Atlantic cod and haddock eggs

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Abstract

Two miniaturized extraction methods for a wide range of 2-6 ring polycyclic aromatic hydrocarbons (PAHs) and their alkylated homologues in small lipid-rich biota samples (≤100 mg) have been developed. Both methods utilize liquid extraction (LE) prior to a clean-up step using either normal phase solid phase extraction (SPE) or mixed-phase dispersive SPE (dSPE). Optimization of the methods was achieved by comparing the type and amount of sorbents, drying agents, and solvents used. In order to improve the limits of detection (LOD) of target PAHs under high sensitivity gas chromatography–tandem mass spectrometry analysis, specific emphasis was given to minimizing lipid co-extraction. The optimized LE–SPE method comprised extraction with dichloromethane/n-hexane (1:1, v/v) and clean-up by silica SPE, whereas the optimized LE–dSPE method comprised extraction with acetonitrile and clean-up with PSA and C18 sorbents. The methods were validated and directly compared through the analysis of Atlantic cod (Gadus morhua) and haddock (Melanogrammus aeglefinus) eggs exposed to oil. The LE–SPE method resulted in lower levels of co-extracted lipids (14.1 ± 1.7 ng/μL) than the LE–dSPE method (60 ± 14 ng/μL). Achieved PAH LODs for the LE–SPE method were typically an order of magnitude lower (<5 ng/g) than for the LE–dSPE method (<125 ng/g). The LE–SPE method offers the possibility for PAH analysis of small samples of fish eggs (~100 mg) exposed to small quantities of crude oil (~1–10 μg/L total PAHs).

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Acknowledgments

This work is part of the project “Assessment of long-term effects of oil exposure on early life stages of haddock”, which is funded by the Norwegian Research Council (grant no. 234367). Marta Silva has received financial support from the Erasmus Mundus program EMQAL. The authors wish to acknowledge Bjørn Grung (University of Bergen) for guidance with experimental designs, Elin Sørhus and Ørjan Karlsen (IMR) for providing samples of fish eggs, and Marianne Unaas Rønsberg (SINTEF) for assistance in the laboratory.

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Authors and Affiliations

  1. Institute of Marine Research, P.O. Box 1870 Nordnes, 5817, Bergen, Norway

    Lisbet Sørensen & Sonnich Meier

  2. Department of Chemistry, University of Bergen, P.O. Box 7800, 5020, Bergen, Norway

    Lisbet Sørensen & Marta S. Silva

  3. SINTEF Materials and Chemistry, P.O. Box 4760 Sluppen, 7465, Trondheim, Norway

    Andy M. Booth

Authors
  1. Lisbet Sørensen
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  2. Marta S. Silva
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  3. Andy M. Booth
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  4. Sonnich Meier
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Corresponding author

Correspondence to Lisbet Sørensen.

Ethics declarations

All animal experiments within the study were approved by NARA, the governmental Norwegian Animal Research Authority (http://www.fdu.no/fdu/, reference number 2012/275334-2). The Austevoll Aquaculture Research Station has the following permission for catch and maintenance of Atlantic haddock: H-AV 77, H-AV 78, and H-AV 79. These are permits given by the Norwegian Directorate of Fisheries. Furthermore, the Austevoll Aquaculture Research Station has a permit to run as a research animal facility using fish (all developmental stages), with code 93 from the national Animal Welfare and Institutional Animal Care and Use Committee (IACUC); NARA.

Conflict of interest

The authors declare no competing interest.

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Sørensen, L., Silva, M.S., Booth, A.M. et al. Optimization and comparison of miniaturized extraction techniques for PAHs from crude oil exposed Atlantic cod and haddock eggs. Anal Bioanal Chem 408, 1023–1032 (2016). https://doi.org/10.1007/s00216-015-9225-x

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  • DOI: https://doi.org/10.1007/s00216-015-9225-x

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