Design and Validation of a High-Throughput Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry Method for Quantification of Hepcidin in Human Plasma

• Published in: Analytical chemistry (Washington) 2011-11, Vol.83 (21), p.8357-8362
• Main Authors: Anderson, Damon S, Kirchner, Marc, Kellogg, Mark, Kalish, Leslie A, Jeong, Jee-Yeong, Vanasse, Gary, Berliner, Nancy, Fleming, Mark D, Steen, Hanno
• Format: Article
• Language: English
• Subjects: Adult; Analytical chemistry; Antimicrobial Cationic Peptides - blood; Chemistry; Child; Desorption; Exact sciences and technology; Female; Hemochromatosis - diagnosis; Hepcidins; High-Throughput Screening Assays; Humans; Immunoassay; Isotopes; Male; Mass spectrometry; Metabolism; Miscellaneous; Plasma; Reference Standards; Spectrometric and optical methods; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/ac2020905

Disorders of iron metabolism affect over a billion people worldwide. The circulating peptide hormone hepcidin, the central regulator of iron distribution in mammals, holds great diagnostic potential for an array of iron-associated disorders, including iron loading (β-thalassemia), iron overload (hereditary hemochromatosis), and iron deficiency diseases. We describe a novel high-throughput matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry assay for quantification of hepcidin in human plasma. This assay involves enrichment using a functionalized MALDI chip, a novel solvent–detergent precipitation buffer, and quantification using a stable isotope labeled internal standard. The linear range of hepcidin in plasma was 1–120 nM, with a low limit of quantification (LOQ) (1 nM), high accuracy (