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Advances in Quantitative Hepcidin Measurements by Time-of-Flight Mass Spectrometry

Assays for the detection of the iron regulatory hormone hepcidin in plasma or urine have not yet been widely available, whereas quantitative comparisons between hepcidin levels in these different matrices were thus far even impossible due to technical restrictions. To circumvent these limitations, w...

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Autores principales: Swinkels, Dorine W., Girelli, Domenico, Laarakkers, Coby, Kroot, Joyce, Campostrini, Natascia, Kemna, Erwin H. J. M., Tjalsma, Harold
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2442656/
https://www.ncbi.nlm.nih.gov/pubmed/18628991
http://dx.doi.org/10.1371/journal.pone.0002706
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author Swinkels, Dorine W.
Girelli, Domenico
Laarakkers, Coby
Kroot, Joyce
Campostrini, Natascia
Kemna, Erwin H. J. M.
Tjalsma, Harold
author_facet Swinkels, Dorine W.
Girelli, Domenico
Laarakkers, Coby
Kroot, Joyce
Campostrini, Natascia
Kemna, Erwin H. J. M.
Tjalsma, Harold
author_sort Swinkels, Dorine W.
collection PubMed
description Assays for the detection of the iron regulatory hormone hepcidin in plasma or urine have not yet been widely available, whereas quantitative comparisons between hepcidin levels in these different matrices were thus far even impossible due to technical restrictions. To circumvent these limitations, we here describe several advances in time-of flight mass spectrometry (TOF MS), the most important of which concerned spiking of a synthetic hepcidin analogue as internal standard into serum and urine samples. This serves both as a control for experimental variation, such as recovery and matrix-dependent ionization and ion suppression, and at the same time allows value assignment to the measured hepcidin peak intensities. The assay improvements were clinically evaluated using samples from various patients groups and its relevance was further underscored by the significant correlation of serum hepcidin levels with serum iron indices in healthy individuals. Most importantly, this approach allowed kinetic studies as illustrated by the paired analyses of serum and urine samples, showing that more than 97% of the freely filtered serum hepcidin can be reabsorbed in the kidney. Thus, the here reported advances in TOF MS-based hepcidin measurements represent critical steps in the accurate quantification of hepcidin in various body fluids and pave the way for clinical studies on the kinetic behavior of hepcidin in both healthy and diseased states.
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spelling pubmed-24426562008-07-16 Advances in Quantitative Hepcidin Measurements by Time-of-Flight Mass Spectrometry Swinkels, Dorine W. Girelli, Domenico Laarakkers, Coby Kroot, Joyce Campostrini, Natascia Kemna, Erwin H. J. M. Tjalsma, Harold PLoS One Research Article Assays for the detection of the iron regulatory hormone hepcidin in plasma or urine have not yet been widely available, whereas quantitative comparisons between hepcidin levels in these different matrices were thus far even impossible due to technical restrictions. To circumvent these limitations, we here describe several advances in time-of flight mass spectrometry (TOF MS), the most important of which concerned spiking of a synthetic hepcidin analogue as internal standard into serum and urine samples. This serves both as a control for experimental variation, such as recovery and matrix-dependent ionization and ion suppression, and at the same time allows value assignment to the measured hepcidin peak intensities. The assay improvements were clinically evaluated using samples from various patients groups and its relevance was further underscored by the significant correlation of serum hepcidin levels with serum iron indices in healthy individuals. Most importantly, this approach allowed kinetic studies as illustrated by the paired analyses of serum and urine samples, showing that more than 97% of the freely filtered serum hepcidin can be reabsorbed in the kidney. Thus, the here reported advances in TOF MS-based hepcidin measurements represent critical steps in the accurate quantification of hepcidin in various body fluids and pave the way for clinical studies on the kinetic behavior of hepcidin in both healthy and diseased states. Public Library of Science 2008-07-16 /pmc/articles/PMC2442656/ /pubmed/18628991 http://dx.doi.org/10.1371/journal.pone.0002706 Text en Swinkels et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Swinkels, Dorine W.
Girelli, Domenico
Laarakkers, Coby
Kroot, Joyce
Campostrini, Natascia
Kemna, Erwin H. J. M.
Tjalsma, Harold
Advances in Quantitative Hepcidin Measurements by Time-of-Flight Mass Spectrometry
title Advances in Quantitative Hepcidin Measurements by Time-of-Flight Mass Spectrometry
title_full Advances in Quantitative Hepcidin Measurements by Time-of-Flight Mass Spectrometry
title_fullStr Advances in Quantitative Hepcidin Measurements by Time-of-Flight Mass Spectrometry
title_full_unstemmed Advances in Quantitative Hepcidin Measurements by Time-of-Flight Mass Spectrometry
title_short Advances in Quantitative Hepcidin Measurements by Time-of-Flight Mass Spectrometry
title_sort advances in quantitative hepcidin measurements by time-of-flight mass spectrometry
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2442656/
https://www.ncbi.nlm.nih.gov/pubmed/18628991
http://dx.doi.org/10.1371/journal.pone.0002706
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