Department of Pathology, Children's Hospital Boston, Boston, Massachusetts 02115, USA.
Anal Chem. 2010 Feb 15;82(4):1551-5. 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 disregulated, 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 matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) mass spectrometry assay for hepcidin in human urine which involves (i) direct enrichment from minute volumes (5 microL) 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 (intraday CV 12.11%; interday CV 13.21%), and a strong correlation upon interlaboratory cross validation with an existing immunoassay. The assay is simple, accurate, and efficient, and the high-throughput performance features of the assay make large-scale clinical research studies feasible.
肽激素铁调素的水平与全身铁状态呈负相关，并且在铁代谢被继发性失调的疾病中增加，例如慢性病贫血。因此，在临床环境中测量铁调素的能力可能对广泛的适应症具有诊断价值。我们描述了一种新颖的定量基质辅助激光解吸电离飞行时间（MALDI-TOF）质谱测定法，用于人尿液中的铁调素，该方法涉及（i）在功能化芯片表面上直接从微量体积（5 μL）的最小处理尿液中富集，（ii）使用稳定同位素标记的内标进行定量，以及（iii）通过 MALDI-TOF 进行分析。性能特点包括宽线性范围（1-1000 nM；LOQ 2.5 nM）、高准确度（90-110%回收率）和精密度（日内 CV 12.11%；日间 CV 13.21%），以及在与现有免疫测定的实验室间交叉验证时具有很强的相关性。该测定方法简单、准确、高效，并且该测定方法的高通量性能特点使得大规模临床研究成为可能。
