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Hydrophilic interaction (HILIC) and reverse phase liquid chromatography (RPLC)–high resolution MS for characterizing lipids profile disruption in serum of anabolic implanted bovines

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Abstract

Glycerophospholipids have been highlighted as the major lipids class affected by trenbolone acetate/estradiol implant administration in bovines. Non-targeted hydrophilic interaction chromatography (HILIC) and reverse phase liquid chromatography (RPLC) coupled to high resolution mass spectrometry have been successfully applied for characterizing lipid profile disruption in serum of implanted bovines. HILIC data pointed towards significant decrease of C22 fatty acids in implanted animals, i.e., C22:3 (p < 0.05), C22:4 (p < 0.05) and C22:6 (p < 0.01), whilst RPLC data confirmed depletion of glycerophospholipids (phosphatidylglycerols, phosphatidylcholine, phosphatidic acid and phosphatidylethanolamine) with C22 fatty acid chains. Using these two complementary methods, complex lipids with the same alkyl chain have been putatively characterized and could further serve to understand the biological mechanism of illicit administration of trenbolone acetate/estradiol implant. These metabolites underpin glycerophospholipid metabolism as potential pathway, which was identified using metabolomic pathway analysis and MetExplore platforms. We hypothesized that implantation of exogenous androgenic and estrogenic steroids induces a depletion of glycerophospholipids with C22 FA chains, by decreasing high-density lipoproteins, the main transporters of glycerophospholipids in plasma.

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Acknowledgments

This work was financially supported by the Italian Ministry of Health (RF-IZV-2008-1175188) and the French Région Pays de la Loire. Dr Judith Kouassi Nzoughet was a fellowship recipient of Région Pays de la Loire (Projet acronym: Metaboscreen, Contract No 2011-12706). Thanks to Merck Animal Health that kindly supplied the ear implants Revalor-XS®. Authors would like to thank Dr Gilles Simard for useful discussion on the biological pathway.

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Authors and Affiliations

  1. Laboratoire d’Étude des Résidus et Contaminants dans les Aliments (LABERCA), ONIRIS, LUNAM Université, 44307, Nantes, France

    J. Kouassi Nzoughet, H. Gallart-Ayala, K. Hennig, G. Dervilly-Pinel & B. Le Bizec

  2. Department of Chemistry, Istituto Zooprofilattico Sperimentale delle Venezie, 35020, Legnaro, Italy

    G. Biancotto

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  1. J. Kouassi Nzoughet
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Correspondence to G. Dervilly-Pinel.

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Research involving Human and Animal Rights

All institutional and national guidelines for the care and use of laboratory animals were followed. Animals were managed according to Directive 86/609/EEC for the protection of animals used for experimental or other scientific purposes, enforced by the Italian D. Lgs No 116 of January 27, 1992 and Directive 63/2010. The animal study was approved by the Animal Experimentation Ethical Committee of the University of Bologna and was therefore performed in accordance with the ethical standards laid down in the 1964 Declaration of Helsinki and its later amendments.

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Nzoughet, J.K., Gallart-Ayala, H., Biancotto, G. et al. Hydrophilic interaction (HILIC) and reverse phase liquid chromatography (RPLC)–high resolution MS for characterizing lipids profile disruption in serum of anabolic implanted bovines. Metabolomics 11, 1884–1895 (2015). https://doi.org/10.1007/s11306-015-0847-y

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