Journal Information
Journal ID (publisher-id): BM
Journal ID (nlm-ta): Biochem Med (Zagreb)
Title: Biochemia Medica
Abbreviated Title: Biochem. Med. (Zagreb)
ISSN (print): 1330-0962
ISSN (electronic): 1846-7482
Publisher: Croatian Society of Medical Biochemistry and Laboratory Medicine
Article Information
Copyright statement: ©Croatian Society of Medical Biochemistry and Laboratory Medicine.
Copyright: 2018, Croatian Society of Medical Biochemistry
License (open-access):
This is an Open Access article distributed under the terms of the Creative Commons Attribution (http://creativecommons.org/licenses/by/4.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Date received: 04 November 2018
Date accepted: 18 January 2019
Publication date (electronic): 15 April 2019
Publication date (print): 15 June 2019
Volume: 29
Issue: 2
Electronic Location Identifier: 020701
Publisher ID: bm-29-2-020701
DOI: 10.11613/BM.2019.020701
Evaluation of a commercial liquid-chromatography high-resolution mass-spectrometry method for the determination of hepcidin-25
Sieglinde Zelzer[2]
Günter Fauler[2]
Markus Herrmann[2]
Author notes:
The first two authors contributed equally to this work.
[*] Corresponding author: enko.dietmar@gmx.at
Introduction
Reliable determination of hepcidin-25, a key regulator of iron metabolism, is important. This study aimed at evaluating the performance of the Hepcidin-25 Liquid Chromatography-Tandem Mass-Spectrometry (LC-MS/MS) Kit (Immundiagnostik AG, Bensheim, Germany) for quantification of the hepcidin-25 protein.
Materials and methods
Precision, accuracy, linearity, and preanalytical requirements of the liquid-chromatography high-resolution mass-spectrometry (LC-HR-MS) method were evaluated. The imprecision and bias acceptance criteria were defined ≤ 15%. We investigated sample stability at room temperature (RT) and after repeated freeze and thaw cycles. Additionally, we assessed serum hepcidin-25 concentrations of 165 healthy adults referred for a medical check-up.
Results
The hepcidin-25 LC-MS/MS assay was linear over the concentration range of 3 – 200 ng/mL. Within- and between-run precision ranged between 1.9 – 8.6% and 5.1 – 12.4%, respectively. The mean bias of the low and high control material was - 2.7% and 2.1%, respectively. At RT, serum samples were stable for 3 h (mean bias + 0.3%). After two and three freeze and thaw cycles, hepcidin-25 concentrations showed a bias of + 8.0 and + 20%, respectively. Of 165 healthy adults, 109 females had a significantly lower median of 8.42 (range: 1.00 – 60.10) ng/mL compared to 56 males with 15.76 (range: 1.50 – 60.50) ng/mL (P = 0.002).
Keywords: hepcidin-25; preanalytical phase; protein biomarker; liquid-chromatography high-resolution mass-spectrometry