Jump to ContentJump to Main Navigation
Show Summary Details
More options …

Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Greaves, Ronda / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter


IMPACT FACTOR 2018: 3.638

CiteScore 2018: 2.44

SCImago Journal Rank (SJR) 2018: 1.191
Source Normalized Impact per Paper (SNIP) 2018: 1.205

Online
ISSN
1437-4331
See all formats and pricing
More options …
Volume 57, Issue 6

Issues

Provisional standardization of hepcidin assays: creating a traceability chain with a primary reference material, candidate reference method and a commutable secondary reference material

Laura E. Diepeveen
  • Department of Laboratory Medicine, Radboud University Medical Center, Nijmegen, The Netherlands
  • Hepcidinanalysis.com, Nijmegen, The Netherlands
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Coby M.M. Laarakkers
  • Department of Laboratory Medicine, Radboud University Medical Center, Nijmegen, The Netherlands
  • Hepcidinanalysis.com, Nijmegen, The Netherlands
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Gustavo Martos
  • Laboratoire National de Métrologie et d’Essais (LNE), Paris, France
  • Departement de Chimie, Bureau International des Poids et Mesures (BIPM), Sèvres, France
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Marta E. Pawlak
  • Department of Laboratory Medicine, Radboud University Medical Center, Nijmegen, The Netherlands
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Fatih F. Uğuz
  • Department of Laboratory Medicine, Radboud University Medical Center, Nijmegen, The Netherlands
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Kim E.S.A. Verberne
  • Department of Laboratory Medicine, Radboud University Medical Center, Nijmegen, The Netherlands
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Rachel P.L. van Swelm
  • Department of Laboratory Medicine, Radboud University Medical Center, Nijmegen, The Netherlands
  • Hepcidinanalysis.com, Nijmegen, The Netherlands
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Siem Klaver
  • Department of Laboratory Medicine, Radboud University Medical Center, Nijmegen, The Netherlands
  • Hepcidinanalysis.com, Nijmegen, The Netherlands
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Anton F.J. de Haan / Kelly R. Pitts / Sukhvinder S. Bansal / Ioana M. Abbas
  • Division 1.5 Protein Analysis, Federal Institute for Materials Research and Testing (BAM), Berlin, Germany
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Marianne Fillet / Thibaud Lefebvre
  • French Center of Porphyria, INSERM U1149, Sorbonne Paris Cité University, Labex GR-Ex, Louis Mourier Hospital APHP, Colombes, France
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Anneke J. Geurts-Moespot
  • Department of Laboratory Medicine, Radboud University Medical Center, Nijmegen, The Netherlands
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Domenico Girelli / Annalisa Castagna / Matthias Herkert / Outi Itkonen / Gordana Olbina / Naohisa Tomosugi / Mark E. Westerman / Vincent Delatour / Cas W. Weykamp / Dorine W. Swinkels
  • Corresponding author
  • Department of Laboratory Medicine, Radboud University Medical Center, Nijmegen, The Netherlands
  • Hepcidinanalysis.com, Nijmegen, The Netherlands
  • Email
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
Published Online: 2018-11-29 | DOI: https://doi.org/10.1515/cclm-2018-0783

Abstract

Background

Hepcidin concentrations measured by various methods differ considerably, complicating interpretation. Here, a previously identified plasma-based candidate secondary reference material (csRM) was modified into a serum-based two-leveled sRM. We validated its functionality to increase the equivalence between methods for international standardization.

Methods

We applied technical procedures developed by the International Consortium for Harmonization of Clinical Laboratory Results. The sRM, consisting of lyophilized serum with cryolyoprotectant, appeared commutable among nine different measurement procedures using 16 native human serum samples in a first round robin (RR1). Harmonization potential of the sRM was simulated in RR1 and evaluated in practice in RR2 among 11 measurement procedures using three native human plasma samples. Comprehensive purity analysis of a candidate primary RM (cpRM) was performed by state of the art procedures. The sRM was value assigned with an isotope dilution mass spectrometry-based candidate reference method calibrated using the certified pRM.

Results

The inter-assay CV without harmonization was 42.1% and 52.8% in RR1 and RR2, respectively. In RR1, simulation of harmonization with sRM resulted in an inter-assay CV of 11.0%, whereas in RR2 calibration with the material resulted in an inter-assay CV of 19.1%. Both the sRM and pRM passed international homogeneity criteria and showed long-term stability. We assigned values to the low (0.95±0.11 nmol/L) and middle concentration (3.75±0.17 nmol/L) calibrators of the sRM.

Conclusions

Standardization of hepcidin is possible with our sRM, which value is assigned by a pRM. We propose the implementation of this material as an international calibrator for hepcidin.

This article offers supplementary material which is provided at the end of the article.

Keywords: harmonization; hepcidin; iron metabolism; primary reference material; secondary reference material; standardization

References

  • 1.

    Nemeth E, Tuttle MS, Powelson J, Vaughn MB, Donovan A, Ward DM, et al. Hepcidin regulates cellular iron efflux by binding to ferroportin and inducing its internalization. Science 2004;306:2090–3.CrossrefPubMedGoogle Scholar

  • 2.

    Girelli D, Nemeth E, Swinkels DW. Hepcidin in the diagnosis of iron disorders. Blood 2016;127:2809–13.CrossrefWeb of ScienceGoogle Scholar

  • 3.

    Kroot JJ, Kemna EH, Bansal SS, Busbridge M, Campostrini N, Girelli D, et al. Results of the first international round robin for the quantification of urinary and plasma hepcidin assays: need for standardization. Haematologica 2009;94:1748–52.PubMedCrossrefWeb of ScienceGoogle Scholar

  • 4.

    Kroot JJ, van Herwaarden AE, Tjalsma H, Jansen RT, Hendriks JC, Swinkels DW. Second round robin for plasma hepcidin methods: first steps toward harmonization. Am J Hematol 2012;87: 977–83.PubMedWeb of ScienceCrossrefGoogle Scholar

  • 5.

    van der Vorm LN, Hendriks JC, Laarakkers CM, Klaver S, Armitage AE, Bamberg A, et al. Toward worldwide hepcidin assay harmonization: identification of a commutable secondary reference material. Clin Chem 2016;62:993–1001.CrossrefPubMedWeb of ScienceGoogle Scholar

  • 6.

    Vesper HW, Myers GL, Miller WG. Current practices and challenges in the standardization and harmonization of clinical laboratory tests. Am J Clin Nutr 2016;104:907S–12S.Google Scholar

  • 7.

    Hoofnagle AN. Harmonization of blood-based indicators of iron status: making the hard work matter. Am J Clin Nutr 2017;106:1615s–9s.Web of ScienceGoogle Scholar

  • 8.

    ISO17511:2003. In vitro diagnostic medical devices – measurement of quantities in biological samples – metrological traceability of values assigned to calibrators and control materials. https://www.iso.org/standard/30716.html. Accessed: 10 Sept 2018.

  • 9.

    Miller WG. Harmonization: its time has come. Clin Chem 2017;63:1184–6.PubMedWeb of ScienceCrossrefGoogle Scholar

  • 10.

    Myers GL, Miller WG. The international consortium for harmonization of clinical laboratory results (ichclr) – a pathway for harmonization. Ejifcc 2016;27:30–6.PubMedGoogle Scholar

  • 11.

    Myers GL, Miller WG. The roadmap for harmonization: status of the international consortium for harmonization of clinical laboratory results. Clin Chem Lab Med 2018;56:1667–72.Web of SciencePubMedCrossrefGoogle Scholar

  • 12.

    Weykamp C, Eckfeld J, Vesper H, Thienpont L, Burns C, Caliendo A, et al. AACC harmonization initiative; toolbox of technical procedures to achieve harmonization for a measurand 2012. www.harmonization.net. Accessed: 10 Sept 2018.

  • 13.

    Bros P, Josephs RD, Stoppacher N, Cazals G, Lehmann S, Hirtz C, et al. Impurity determination for hepcidin by liquid chromatography-high resolution and ion mobility mass spectrometry for the value assignment of candidate primary calibrators. Anal Bioanal Chem 2017;409:2559–67.Web of SciencePubMedCrossrefGoogle Scholar

  • 14.

    Laarakkers CM, Wiegerinck ET, Klaver S, Kolodziejczyk M, Gille H, Hohlbaum AM, et al. Improved mass spectrometry assay for plasma hepcidin: detection and characterization of a novel hepcidin isoform. PLoS One 2013;8:e75518.CrossrefPubMedWeb of ScienceGoogle Scholar

  • 15.

    Pechlaner R, Kiechl S, Mayr M, Santer P, Weger S, Haschka D, et al. Correlates of serum hepcidin levels and its association with cardiovascular disease in an elderly general population. Clin Chem Lab Med 2016;54:151–61.Web of ScienceGoogle Scholar

  • 16.

    Murao N, Ishigai M, Yasuno H, Shimonaka Y, Aso Y. Simple and sensitive quantification of bioactive peptides in biological matrices using liquid chromatography/selected reaction monitoring mass spectrometry coupled with trichloroacetic acid clean-up. Rapid Commun Mass Spectrom 2007;21:4033–8.CrossrefPubMedWeb of ScienceGoogle Scholar

  • 17.

    Udali S, Castagna A, Corbella M, Ruzzenente A, Moruzzi S, Mazzi F, et al. Hepcidin and DNA promoter methylation in hepatocellular carcinoma. Eur J Clin Invest 2018;48.Web of SciencePubMedGoogle Scholar

  • 18.

    Itkonen O, Parkkinen J, Stenman UH, Hamalainen E. Preanalytical factors and reference intervals for serum hepcidin lc-ms/ms method. Clin Chim Acta 2012;413:696–701.Web of SciencePubMedCrossrefGoogle Scholar

  • 19.

    Lefebvre T, Dessendier N, Houamel D, Ialy-Radio N, Kannengiesser C, Manceau H, et al. Lc-ms/ms method for hepcidin-25 measurement in human and mouse serum: Clinical and research implications in iron disorders. Clin Chem Lab Med 2015;53:1557–67.Web of SciencePubMedGoogle Scholar

  • 20.

    Abbas IM, Hoffmann H, Montes-Bayon M, Weller MG. Improved lc-ms/ms method for the quantification of hepcidin-25 in clinical samples. Anal Bioanal Chem 2018;410:3835–46.PubMedWeb of ScienceCrossrefGoogle Scholar

  • 21.

    Kroot JJ, Laarakkers CM, Geurts-Moespot AJ, Grebenchtchikov N, Pickkers P, van Ede AE, et al. Immunochemical and mass-spectrometry-based serum hepcidin assays for iron metabolism disorders. Clin Chem 2010;56:1570–9.CrossrefPubMedWeb of ScienceGoogle Scholar

  • 22.

    Zipperer E, Post JG, Herkert M, Kundgen A, Fox F, Haas R, et al. Serum hepcidin measured with an improved elisa correlates with parameters of iron metabolism in patients with myelodysplastic syndrome. Ann Hematol 2013;92:1617–23.Web of SciencePubMedCrossrefGoogle Scholar

  • 23.

    Butterfield AM, Luan P, Witcher DR, Manetta J, Murphy AT, Wroblewski VJ, et al. A dual-monoclonal sandwich elisa specific for hepcidin-25. Clin Chem 2010;56:1725–32.CrossrefPubMedWeb of ScienceGoogle Scholar

  • 24.

    Budd JR, Weykamp C, Rej R, MacKenzie F, Ceriotti F, Greenberg N, et al. IFCC working group recommendations for assessing commutability part 3: using the calibration effectiveness of a reference material. Clin Chem 2018;64:465–74.CrossrefWeb of ScienceGoogle Scholar

  • 25.

    Fraser CG, Hyltoft Petersen P, Libeer JC, Ricos C. Proposals for setting generally applicable quality goals solely based on biology. Ann Clin Biochem 1997;34:8–12.CrossrefPubMedGoogle Scholar

  • 26.

    Murphy AT, Witcher DR, Luan P, Wroblewski VJ. Quantitation of hepcidin from human and mouse serum using liquid chromatography tandem mass spectrometry. Blood 2007;110:1048–54.PubMedWeb of ScienceCrossrefGoogle Scholar

  • 27.

    ISO13528:2005. Statistical methods for use in proficiency testing by interlaboratory comparisons. https://www.iso.org/standard/35664.html. Accessed: 10 Sept 2018.

  • 28.

    ISO/IECGuide98-3:2008. Uncertainty of measurement – part 3: guide to the expression of uncertainty in measurement (gum:1995). https://www.iso.org/standard/50461.html. Accessed: 10 Sept 2018.

  • 29.

    Kroot JJ, Hendriks JC, Laarakkers CM, Klaver SM, Kemna EH, Tjalsma H, et al. (Pre)analytical imprecision, between-subject variability, and daily variations in serum and urine hepcidin: implications for clinical studies. Anal Biochem 2009; 389:124–9.CrossrefPubMedWeb of ScienceGoogle Scholar

  • 30.

    Schaap CC, Hendriks JC, Kortman GA, Klaver SM, Kroot JJ, Laarakkers CM, et al. Diurnal rhythm rather than dietary iron mediates daily hepcidin variations. Clin Chem 2013; 59:527–35.PubMedCrossrefWeb of ScienceGoogle Scholar

  • 31.

    Kemna E, Tjalsma H, Laarakkers C, Nemeth E, Willems H, Swinkels D. Novel urine hepcidin assay by mass spectrometry. Blood 2005;106:3268–70.CrossrefPubMedGoogle Scholar

  • 32.

    Ganz T, Olbina G, Girelli D, Nemeth E, Westerman M. Immunoassay for human serum hepcidin. Blood 2008;112:4292–7.CrossrefWeb of SciencePubMedGoogle Scholar

About the article

Corresponding author: Prof. Dr. Dorine W. Swinkels, Department of Laboratory Medicine, Translational Metabolic Laboratory (830), Radboud University Medical Center, P.O. Box 9101, 6500 HB Nijmegen, The Netherlands; and Hepcidinanalysis.com, Nijmegen, The Netherlands, Phone: +31 (0)24-3618957, Fax: +31 (0)24-3668754


Received: 2018-07-23

Accepted: 2018-10-04

Published Online: 2018-11-29

Published in Print: 2019-05-27


Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

Research funding: We would like to acknowledge Fondazione Cariverona. The funding organization played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

Employment or leadership: LD, CL, SK, RvS and DS are employees of Radboudumc, that via its Hepcidinanalysis.com initiative offers high quality hepcidin measurements to the medical, scientific and pharmaceutical community at a fee for the service basis. KRP is an employee of Corgenix Medical Corporation specializing in IVD development, manufacturing, and distribution. MW (deceased) was President and CEO of Intrinsic LifeSciences LLC. GO is an employee of Intrinsic LifeSciences LLC and has ownership interest in the company. Intrinsic LifeSciences is engaged in the commercial development of hepcidin and related diagnostic products and provides CLIA compliant hepcidin testing to research, clinical and pharmaceutical clients worldwide.

Honorarium: None declared.

Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.


Citation Information: Clinical Chemistry and Laboratory Medicine (CCLM), Volume 57, Issue 6, Pages 864–872, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/cclm-2018-0783.

Export Citation

©2019 Walter de Gruyter GmbH, Berlin/Boston.Get Permission

Supplementary Article Materials

Comments (0)

Please log in or register to comment.
Log in