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Measurement of hepcidin isoforms in human serum by liquid chromatography with high resolution mass spectrometry

Research output: Contribution to journalArticle

Original languageEnglish
Pages (from-to)541-553
Number of pages13
Issue number6
Early online date23 Feb 2017
Accepted/In press19 Jan 2017
E-pub ahead of print23 Feb 2017
PublishedMar 2017


King's Authors


AIM: Hepcidin-25 is the master regulator of iron homeostasis. N-truncated isoforms of hepcidin-25 have been identified (hepcidin-20, -22, -24), although data on the concentrations of these isoforms are sparse.

MATERIALS & METHODS: Serum was mixed with aqueous formic acid, and the supernatant loaded onto a 96-well-SPE-plate. Eluted analytes were analyzed using LC-HR-MS. Forty-seven paired dipotassium-EDTA human plasma and serum samples were analyzed.

RESULTS: The LLOQ was 1 μg/l (all analytes). Accuracy and precision were acceptable. There was a good correlation (R(2) >0.90, all analytes) between matrices. The median (range) serum hepcidin-20, -22, -24 and -25 concentrations measured were 4 (1-40), 8 (2-20), 8 (1-50) and 39 (1-334) μg/l, respectively.

CONCLUSION: LC-HR-MS is widely applicable to the measurement of hepcidin-25, and truncated isoforms.

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