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Impurity determination for hepcidin by liquid chromatography-high resolution and ion mobility mass spectrometry for the value assignment of candidate primary calibrators

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Abstract

In metrology institutes, the state-of-the-art for purity analysis of peptides/proteins mainly addresses short and unfolded peptides. Important developments are anticipated for the characterization of nonlinear peptides or proteins. Hepcidin 1-25 is an interesting model system because this small protein contains four disulfide bridges with a particular connectivity that is difficult to reproduce and could induce a bias in quantification. Hepcidin 1-25 is involved in iron-related disorders and anemia, in an inflammatory context, and its clinical relevance in neurodegenerative disorders is under investigation. It is also an emerging biomarker. Recent inter-laboratory studies showed a need for standardization of hepcidin assay and the need to produce certified reference materials. This paper discusses two hepcidin standards from different synthesis pathways that have been characterized by high-resolution mass spectrometry and ion mobility mass spectrometry.

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Abbreviations

hrMS:

High-resolution mass spectrometry

IM-MS:

Ion mobility mass spectrometry

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Acknowledgements

The authors would like to thank Maria Colombo from Eurogentec for making available different batches of hepcidin material and for her support of the study design.

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Authors and Affiliations

  1. Laboratoire National de Métrologie et d’Essais (LNE), 1 Rue Gaston Boissier, 75015, Paris, France

    Pauline Bros & Vincent Delatour

  2. Departement de Chimie, Bureau International des Poids et Mesures (BIPM), Pavillon de Breteuil, 92312, Sèvres, France

    Pauline Bros, Ralf D. Josephs, Norbert Stoppacher & Robert I. Wielgosz

  3. Laboratoire de Biochimie et de Protéomique Clinique – Institut de Médecine Régénérative et Biothérapies (LBPC-IRMB), CHU de Montpellier, 80 rue Augustin Fliche, 34091, Montpellier, France

    Pauline Bros, Sylvain Lehmann & Christophe Hirtz

  4. Laboratoire de Mesures Physiques, Place Eugène Bataillon, 34095, Montpellier, France

    Guillaume Cazals

Authors
  1. Pauline Bros
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  2. Ralf D. Josephs
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  3. Norbert Stoppacher
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  4. Guillaume Cazals
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  5. Sylvain Lehmann
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  6. Christophe Hirtz
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  7. Robert I. Wielgosz
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  8. Vincent Delatour
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Corresponding author

Correspondence to Ralf D. Josephs.

Ethics declarations

The experiments were conducted with synthetic hepcidin materials provided by Eurogentec (Seraing, Belgium).

Conflict of interest

The authors declare that they have no conflict of interest.

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Bros, P., Josephs, R.D., Stoppacher, N. et al. Impurity determination for hepcidin by liquid chromatography-high resolution and ion mobility mass spectrometry for the value assignment of candidate primary calibrators. Anal Bioanal Chem 409, 2559–2567 (2017). https://doi.org/10.1007/s00216-017-0202-4

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  • DOI: https://doi.org/10.1007/s00216-017-0202-4

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