title: LC-MS/MS method for hepcidin-25 measurement in human and mouse serum: clinical and research implications in iron disorders.
abstract: BACKGROUND
  The peptide hepcidin plays a central role in regulating dietary iron absorption and body iron distribution. This 25-amino acid hormone is produced and secreted predominantly by hepatocytes. Hepcidin has been suggested as a promising diagnostic marker for iron-related disorders. However, its accurate quantification for clinical use remains so far challenging. In this report we describe a highly specific and quantitative serum hepcidin method using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS).
  
  
  METHODS
  The analytical validation included the determination of the limit of detection, of quantification, repeatability, reproducibility and linearity. This assay was developed for human and mouse hepcidin. The human assay was performed on serum patients with unexplained microcytic anemia. We applied our LC-MS/MS method for quantifying hepcidin-1 in mouse in various conditions: inflammation, hemolytic anemia, Hamp-1, Hjv and Hfe KO mice.
  
  
  RESULTS
  We show that the LC-MS/MS is suitable for accurate determination of hepcidin-25 in clinical samples, thereby representing a useful tool for the clinical diagnosis and follow-up of iron-related diseases. In mouse, a strong correlation between hepatic Hamp-1 mRNA expression and serum hepcidin-1 levels was found (r=0.88; p=0.0002) and the expected variations in mouse models of iron disorders were observed.
  
  
  CONCLUSIONS
  Therefore, we propose this adaptive LC-MS/MS method as a suitable method for accurate determination of hepcidin-25 in clinical samples and as a major tool contributing to the clinical diagnosis, follow-up and management of iron-related disorders. It also opens new avenues to measure hepcidin in animal models without interspecies antigenic limitations.
authors.forename: Thibaud
authors.initials: T
authors.lastname: Lefebvre
authors.forename: Nathalie
authors.initials: N
authors.lastname: Dessendier
authors.forename: Dounia
authors.initials: D
authors.lastname: Houamel
authors.forename: Nathalie
authors.initials: N
authors.lastname: Ialy-Radio
authors.forename: Caroline
authors.initials: C
authors.lastname: Kannengiesser
authors.forename: Hana
authors.initials: H
authors.lastname: Manceau
authors.forename: Carole
authors.initials: C
authors.lastname: Beaumont
authors.forename: Gael
authors.initials: G
authors.lastname: Nicolas
authors.forename: Laurent
authors.initials: L
authors.lastname: Gouya
authors.forename: Hervé
authors.initials: H
authors.lastname: Puy
authors.forename: Zoubida
authors.initials: Z
authors.lastname: Karim
mesh_terms: D000328:Adult; D000595:Amino Acid Sequence; D018798:Anemia, Iron-Deficiency / Q000097:blood*; D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D005260:Female; D064451:Hepcidins / Q000097:blood*; D006801:Humans; D007501:Iron / Q000097:blood; D008297:Male; D051379:Mice; D008875:Middle Aged; D023421:Models, Animal; D008969:Molecular Sequence Data; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*; D055815:Young Adult
chemical_list: C579599:Hamp protein, mouse; D064451:Hepcidins; C545475:hepcidin 25, human; D007501:Iron