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A two-dimensional HPLC separation for the enantioselective determination of hexabromocyclododecane (HBCD) isomers in biota samples

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Abstract

A new method for enantioselective analysis of isomers of hexabromocyclododecane (HBCD) is described, using a two-dimensional high-performance liquid chromatography (HPLC) approach to avoid coelution, in particular between (+) α-HBCD, (+) β-HBCD, or (+) γ-HBCD. After isomer separation on a conventional column, the single isomers are transferred to an enantioselective HPLC column using heart cuts. Two enantioseparations are conducted in two separate partial chromatograms: one for α-HBCD and one for β- and γ-HBCD. The result is a completely undisturbed enantioselective separation for α-HBCD at a resolution of 4.11. A peak capacity of 107 was achieved. This peak capacity is utilized by the six peaks of the three isomers with two enantiomers each by 6 %. This method was applied to samples of sand eel oil, glaucous gull, and ringed seal. The calibration was performed by treating each enantiomer as a single analyte using a multilevel internal standard calibration. Enantiomeric fractions of 0.495–0.501 with standard deviations (SDs) of 0.056–0.071 were determined for racemic standards of α-HBCD, while the values for fish oil were 0.548–0.562 with SD of 0.018–0.041, depending on the respective mass spectrometric transition.

Enantioseparation of a HBCD in a 2-D separation

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Acknowledgments

Thanks to Birgit Groth, who performed the sample preparation and integration of large datasets for this project as a lab technician. The glaucous gull sample originates from the Arctic Monitoring and Assessment Programme funded by the Danish Environmental Protection Agency and coordinated by Frank F. Rigét (Institute of Bioscience, Aarhus University, Denmark).

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  1. Department of Environmental Science, Aarhus University, Frederiksborgsvej 399, 4000, Roskilde, Denmark

    Kai Bester & Katrin Vorkamp

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  1. Kai Bester
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Correspondence to Kai Bester.

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Bester, K., Vorkamp, K. A two-dimensional HPLC separation for the enantioselective determination of hexabromocyclododecane (HBCD) isomers in biota samples. Anal Bioanal Chem 405, 6519–6527 (2013). https://doi.org/10.1007/s00216-013-7100-1

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  • DOI: https://doi.org/10.1007/s00216-013-7100-1

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