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Speciation of Serum Copper and Zinc-Binding High- and Low-Molecular Mass Ligands in Dairy Cows Using HPLC-ICP-MS Technique

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Abstract

The objective of the present study was assessment of the major copper and zinc species in dairy cow blood serum using a hybrid high-performance liquid chromatography-inductively coupled plasma-mass spectrometry (HPLC-ICP-MS) technique. A total of seventeen 5–6-year-old female Simmental cows, cultivated in the Southern Ural region, were examined. Speciation of serum Cu and Zn was performed using chromatographic PerkinElmer Series 200 system equipped with Agilent Bio SEC-5 Column and docked with NexION 300D mass spectrometer. Analysis of serum 63Cu species revealed four major fractions containing 2.5% (A), 15.6% (B), 75.6% (C), and 11.9% (D) of total copper levels. The revealed fractions could be assigned to tetrameric and dimeric macroglobulin, ceruloplasmin, albumin, and low molecular mass (LMM) copper compounds, respectively. Minor fraction (E) containing <1% of total serum Cu levels may be represented by low-molecular mass Cu species. Speciation analysis also revealed four Zn fractions containing 6.3% (A), 16.9% (B), 71% (C), and 3% (D) of total Zn levels that may be attributed to zinc-bound tetrameric and dimeric macroglobulin, albumin, and Zn-amino acid compounds. Correlation analysis demonstrated that relative levels (%) of Zn-B (dimeric α2-macroglobulin), Zn-C (albumin), and Zn-D (LMM) fractions correlate inversely with Cu-A (monomeric α2-macroglobulin) (r = −0.600), Cu-D (albumin) (r = −0.696), and Cu-C (ceruloplasmin) (r = −0.652), respectively. The obtained data demonstrate the particular features of Zn and Cu transport in dairy cows that may be used for assessment of dietary status of trace elements.

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Acknowledgements

The study was performed in agreement with the plan of the studies of FSBSI FSC BST RAS № FNWZ-2019-0001.

Funding

The study was performed with the financial support of the Russian Science Foundation (project No. 078).

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Authors and Affiliations

  1. Orenburg State University, Orenburg, 460018, Russia

    Sergey A. Miroshnikov, Svetlana V. Notova, Anatoly V. Skalny & Alexey A. Tinkov

  2. Federal Research Centre of Biological Systems and Agrotechnologies of the Russian Academy of Sciences, Orenburg, 460000, Russia

    Svetlana V. Notova, Elena A. Sizova, Olga V. Marshinskaia, Tatiana V. Kazakova, Anatoly V. Skalny, Olga P. Ajsuvakova & Alexey A. Tinkov

  3. Micronutrients LTD, Moscow, 105082, Russia

    Margarita G. Skalnaya

  4. Helmholtz Center Munich, D-85764, Neuherberg, Germany

    Bernhard Michalke

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  1. Sergey A. Miroshnikov
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  2. Svetlana V. Notova
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  3. Margarita G. Skalnaya
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Fig. S1.

SEC-ICP-MS chromatograms of the copper blood serum species of the same sample obtained during the initial and repeated sampling (PNG 194 kb)

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Miroshnikov, S.A., Notova, S.V., Skalnaya, M.G. et al. Speciation of Serum Copper and Zinc-Binding High- and Low-Molecular Mass Ligands in Dairy Cows Using HPLC-ICP-MS Technique. Biol Trace Elem Res 200, 591–599 (2022). https://doi.org/10.1007/s12011-021-02666-6

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