A high throughput MALDI-TOF mass spectrometry method for quantification of hepcidin in human urine

• Published in: Analytical chemistry (Washington) 2010-02, Vol.82 (4), p.1551-1555
• Main Authors: Anderson, Damon S., Heeney, Matthew M., Roth, Udo, Menzel, Christoph, Fleming, Mark D., Steen, Hanno
• Format: Article
• Language: English
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/ac902479p

Levels of the peptide hormone hepcidin negatively correlate with systemic iron status and are increased in disorders in which iron metabolism is secondarily dysregulated, such as the anemia of chronic disease. Consequently, the ability to measure hepcidin in the clinical setting may have diagnostic value for a broad range of indications. We describe a novel quantitative MALDI-TOF mass spectrometry assay for hepcidin in human urine which involves, i) direct enrichment from minute volumes (5 μL) of minimally treated urine on the surface of a functionalized chip, ii) quantification by the use of a stable isotope labeled internal standard and iii) analysis by MALDI-TOF. Performance features include: a wide linear range (1–1000 nM; LOQ 2.5 nM), high accuracy (90–110% recovery) and precision (intra-day CV 12.11%; inter-day CV 13.21%), and a strong correlation upon inter-laboratory cross validation with an existing immunoassay. The assay is simple, accurate, efficient, and the high throughput performance features of the assay make large-scale clinical research studies feasible.