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Design and validation of a novel high-throughput Matrix-Assisted Laser Desorption Ionization Time-of-Flight mass spectrometry method for quantification of hepcidin in human plasma. Anal Chem. 2011 Sep 29; Authors: Anderson DS, Kirchner M, Kellogg M, Kalish L, Jeong JY, Vanasse G, Berliner N, Fleming MD, Steen H Abstract
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 LOQ (1nM), high accuracy (<15 %RE), and high precision (intraday average 5.52-18.48 %CV and interday 9.32-14.83 %CV). The assay showed strong correlation with an established hepcidin immunoassay (Spearman; R2=0.839 n=93 EDTA plasma). A collection of normal healthy pediatric samples (range 3.8 - 32.5 ng/mL; mean 12.9 ng/mL; n=119) showed significant differences from an adult collection (range 1.8 - 48.7 ng/mL; mean 16.1 ng/mL; n=95; P=0.0096). We discuss these preliminary reference ranges and correlations with additional parameters in light of the utility and limitations of hepcidin measurements as a stand-alone diagnostic and as a tool for therapeutic intervention.
PMID: 21958231 [PubMed - as supplied by publisher] Read more |
