Original scientific paper

https://doi.org/10.11613/BM.2019.020701

Evaluation of a commercial liquid-chromatography high-resolution mass-spectrometry method for the determination of hepcidin-25

Dietmar Enko orcid.org/0000-0002-1173-0928 ; Institute of Clinical Chemistry and Laboratory Medicine, General Hospital Steyr, Steyr, Austria; Clinical Institute of Medical and Chemical Laboratory Diagnostics, Medical University of Graz, Graz, Austria
Sieglinde Zelzer ; Clinical Institute of Medical and Chemical Laboratory Diagnostics, Medical University of Graz, Graz, Austria
Günter Fauler ; Clinical Institute of Medical and Chemical Laboratory Diagnostics, Medical University of Graz, Graz, Austria
Markus Herrmann ; Clinical Institute of Medical and Chemical Laboratory Diagnostics, Medical University of Graz, Graz, Austria

Abstract

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 massspectrometry (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).
Conclusions: The hepcidin-25 LC-MS/MS kit shows a broad analytical range and meets the imprecision and bias acceptance criteria of ≤ 15%. Serum samples can be stored at RT for 3 h and resist up to two freeze and thaw cycles.

Keywords

hepcidin-25; preanalytical phase; protein biomarker; liquid-chromatography high-resolution mass-spectrometry

Hrčak ID:

218971

URI

https://hrcak.srce.hr/218971

Publication date:

15.6.2019.

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