title: Validation of hepcidin quantification in plasma using LC-HRMS and discovery of a new hepcidin isoform.
abstract: BACKGROUND
  Hepcidin, a 25 amino acid peptide, plays an important role in iron homeostasis. Some hepcidin truncated peptides have antibiotic effects.
  
  
  RESULTS
  A new analytical method for hepcidin determination in human plasma using LC-HRMS operating in full-scan acquisition mode has been validated. The extraction consists of protein precipitation and a drying reconstitution step; a 2.1 x 50 mm (idxL) C18 analytical column was used. Detection specificity, stability, accuracy, precision and recoveries were determined. The LOQ/LOD were 0.25/0.1 nM, respectively. More than 600 injections of plasma extracts were performed, allowing evaluation of the assay robustness. Hepcidin-20, hepcidin-22 and a new isoform, hepcidin-24, were detected in patients.
  
  
  CONCLUSIONS
  The data underscore the usefulness of LC-HRMS for in-depth investigations related to hepcidin levels and pathways.
authors.affiliation: Quantitative Mass Spectrometry Facility University of Lausanne -CHUV, Route du Bugnon, BH18-218, 1011 Lausanne, Switzerland.
authors.forename: Bertrand
authors.initials: B
authors.lastname: Rochat
authors.forename: Davide
authors.initials: D
authors.lastname: Peduzzi
authors.forename: Justin
authors.initials: J
authors.lastname: McMullen
authors.forename: Amélie
authors.initials: A
authors.lastname: Favre
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Kottelat
authors.forename: Bernard
authors.initials: B
authors.lastname: Favrat
authors.forename: Jean-Daniel
authors.initials: JD
authors.lastname: Tissot
authors.forename: Anne
authors.initials: A
authors.lastname: Angelillo-Scherrer
authors.forename: Maciej
authors.initials: M
authors.lastname: Bromirski
authors.forename: Sophie
authors.initials: S
authors.lastname: Waldvogel
mesh_terms: D000595:Amino Acid Sequence; D002138:Calibration; D002853:Chromatography, Liquid / Q000592:standards*; D005260:Female; D064451:Hepcidins / Q000097:blood*; D006801:Humans; D008297:Male; D013058:Mass Spectrometry / Q000379:methods / Q000592:standards*; D008875:Middle Aged; D008969:Molecular Sequence Data; D020033:Protein Isoforms / Q000097:blood; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D052617:Solid Phase Microextraction
chemical_list: D064451:Hepcidins; D020033:Protein Isoforms