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Quantitation of hepcidin in serum using ultra‐high‐pressure liquid chromatography and a linear ion trap mass spectrometer
Author(s) -
Bansal Sukhvinder S.,
Abbate Vincenzo,
Bomford Adrian,
Halket John M.,
Macdougall Iain C.,
Thein Swee Lay,
Hider Robert C.
Publication year - 2010
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.4512
Subject(s) - chemistry , hepcidin , chromatography , hereditary hemochromatosis , detection limit , mass spectrometry , quadrupole ion trap , ion trap , hemochromatosis , medicine , inflammation
Hepcidin is a peptide hormone that functions as a key regulator of mammalian iron metabolism. Biological levels are increased in end‐stage renal disease and during inflammation but suppressed in hemochromatosis. Thus hepcidin levels have diagnostic importance. This study describes the development of an analytical method for the quantitative determination of the concentration of hepcidin in clinical samples. The fragmentation of hepcidin was investigated using triple quadrupole and linear ion trap mass spectrometers. A standard quantity of a stable isotopically labelled hepcidin internal standard was added to serum samples. Extraction was performed by protein precipitation and weak cation‐exchange magnetic nanoparticles. Chromatography was carried out on sub 2 µm particle stationary phase, using ultra‐high‐pressure liquid chromatography and a linear ion trap for quantitation. The lower limit of quantitation was 0.4 nmol/L with less than 20% accuracy and precision. The mean hepcidin concentration in sera for controls was 4.6 ± 2.7 nmol/L, in patients with sickle cell disease, 7.0 ± 8.9 nmol/L; in patients with end‐stage renal disease, 30.5 ± 15.7 nmol/L; and patients with penetrant hereditary hemochromatosis, 1.4 ± 0.8 nmol/L. Copyright © 2010 John Wiley & Sons, Ltd.