Mass spectrometry measurement of plasma hepcidin for the prediction of iron overload

Annarosa Floreani, Filippo Navaglia, Erik Rosa Rizzotto, Daniela Basso, Maria Chiaramonte, Andrea Padoan, Ioannis Petridis, Nora Cazzagon, Roberto Testa, Maurizio Marra, Mario Plebani, Osvaldo Di Andrea, Salvatore Lobello, Barbara Marracini, Isabella Carderi, Emanuele Lattanzi

Research output: Contribution to journalArticlepeer-review


Background: Hepcidin has emerged as the primary regulator of iron homeostasis. Previous studies on assessing urinary hepcidin are limited. We developed a method for quantifying hepcidin-25 (Hep-25) in plasma using surface-enhanced laser-desorption-ionization time-of-flight mass spectrometry (SELDI-TOF/MS) and a 25-AA peptide as reference standard. The aims of the study were 1) to assess the performance of this method in different conditions of iron metabolism disorders; 2) to assess the diagnostic validity of non-invasive serum biomarkers in the identification of iron overload. Methods: Validation of the method was performed in 10 patients with type I hemochromatosis (HE) and in 177 subjects previously enrolled in a general population epidemiological study. Among the latter group, 17 had non-alcoholic fatty liver disease, 10 had chronic hepatitis C, and 150 subjects had normal ultrasound, normal liver function tests (LFTs), an alcohol intake 4.8 mmol/L and Hep-25/SF ratio ≤6.6 as the main splitters. These variables enabled the correct diagnosis of HE with 100% sensitivity, 93% specificity and an area under the receiver operating characteristic (ROC) curve of 0.993. Conclusions: Our plasma Hep-25 mass spectrometry method yields measurements that reflect pathological and genetic influences; simple non-invasive biomarkers (Hep-25/SF ratio and glucose) can predict the presence of HE.

Original languageEnglish
Pages (from-to)197-206
Number of pages10
JournalClinical Chemistry and Laboratory Medicine
Issue number2
Publication statusPublished - Feb 1 2011


  • hemochromatosis
  • hepcidin
  • iron overload
  • surface-enhanced laser-desorption-ionization time-of-flight mass spectrometry (SELDI-TOF/MS)

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, medical


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