title: Impurity determination for hepcidin by liquid chromatography-high resolution and ion mobility mass spectrometry for the value assignment of candidate primary calibrators.
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.
authors.affiliation: Laboratoire National de Métrologie et d'Essais (LNE), 1 Rue Gaston Boissier, 75015, Paris, France.|Departement de Chimie, Bureau International des Poids et Mesures (BIPM), Pavillon de Breteuil, 92312, Sèvres, France.|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.
authors.forename: Pauline
authors.initials: P
authors.lastname: Bros
authors.affiliation: Departement de Chimie, Bureau International des Poids et Mesures (BIPM), Pavillon de Breteuil, 92312, Sèvres, France. ralf.josephs@bipm.org.
authors.forename: Ralf D
authors.initials: RD
authors.lastname: Josephs
authors.affiliation: Departement de Chimie, Bureau International des Poids et Mesures (BIPM), Pavillon de Breteuil, 92312, Sèvres, France.
authors.forename: Norbert
authors.initials: N
authors.lastname: Stoppacher
authors.affiliation: Laboratoire de Mesures Physiques, Place Eugène Bataillon, 34095, Montpellier, France.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Cazals
authors.affiliation: 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.
authors.forename: Sylvain
authors.initials: S
authors.lastname: Lehmann
authors.affiliation: 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.
authors.forename: Christophe
authors.initials: C
authors.lastname: Hirtz
authors.affiliation: Departement de Chimie, Bureau International des Poids et Mesures (BIPM), Pavillon de Breteuil, 92312, Sèvres, France.
authors.forename: Robert I
authors.initials: RI
authors.lastname: Wielgosz
authors.affiliation: Laboratoire National de Métrologie et d'Essais (LNE), 1 Rue Gaston Boissier, 75015, Paris, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Delatour
mesh_terms: D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods* / Q000592:standards*; D004340:Drug Contamination / Q000517:prevention & control*; D064451:Hepcidins / Q000032:analysis* / Q000302:isolation & purification*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods* / Q000592:standards*; D012015:Reference Standards
chemical_list: D064451:Hepcidins
keywords: Disulfide bridges; Hepcidin; High-resolution mass spectrometry; Ion mobility, conformers; Purity assessment