title: Hepcidin-25: Measurement by LC-MS/MS in serum and urine, reference ranges and urinary fractional excretion.
abstract: A liquid chromatography tandem-mass spectrometry (LC-MS/MS) method was developed for reliably quantifying hepcidin-25 in human urine and serum. A 95% reference range was established for serum hepcidin-25 levels by standardizing the sampling time between 8:00am and 11:00am in 90 apparently healthy volunteers. The association between hepcidin-25 concentration and other biological parameters was studied using multivariable analysis and the coefficient of renal excretion of hepcidin-25 was calculated. Preanalytical variables were also investigated. The LC-MS/MS method was validated using a recent validation strategy based on accuracy profiles. Good results were obtained in terms of trueness, precision, and linearity in the following dosing ranges: from 0.77 to 200nmol/L for urine and from 0.48 to 100nmol/L for serum. The 95% reference range of serum hepcidin-25 concentration established after excluding known conditions that affect hepcidin-25 expression was 1.5 to 15.2nmol/L. A difference between genders was demonstrated with a median concentration of 5.5 versus 7.2nmol/L for women and men, respectively. Serum hepcidin-25 concentrations were strongly correlated with ferritin and, to a lesser extent, with iron levels. The coefficient of renal excretion ranged from 0.1 to 16.4%. Higher values of hepcidin-25 concentrations were observed on ethylene diamine tetraacetate tubes compared to serum or lithium-heparin devices.
authors: Department of Clinical Chemistry, Hôpital Erasme, Belgium. fleur.wolff@erasme.ulb.ac.be
authors: Fleur
authors: F
authors: Wolff
authors: Marie
authors: M
authors: Deleers
authors: Christian
authors: C
authors: Melot
authors: Béatrice
authors: B
authors: Gulbis
authors: Frédéric
authors: F
authors: Cotton
mesh_terms: D000328:Adult; D018798:Anemia, Iron-Deficiency / Q000097:blood; D001774:Blood Chemical Analysis / Q000379:methods*; D002853:Chromatography, Liquid; D005260:Female; D064451:Hepcidins / Q000097:blood* / Q000652:urine*; D006801:Humans; D007501:Iron / Q000097:blood; D008297:Male; D008875:Middle Aged; D012016:Reference Values; D053719:Tandem Mass Spectrometry; D016482:Urinalysis / Q000379:methods*; D055815:Young Adult
chemical_list: D064451:Hepcidins; D007501:Iron