advances in quantitative hepcidin measurements by time-of-flight mass spectrometry飞行时间质谱定量hepcidin进步测量.pdfVIP

Advances in Quantitative Hepcidin Measurements by Time-of-Flight Mass Spectrometry 1 2 1 1 2 Dorine W. Swinkels *, Domenico Girelli , Coby Laarakkers , Joyce Kroot , Natascia Campostrini , Erwin H. J. M. Kemna1, Harold Tjalsma1 1 Department of Clinical Chemistry, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands, 2 Department of Clinical and Experimental Medicine, Section of Internal Medicine, University of Verona, Verona, Italy Abstract Assays for the detection of the iron regulatory hormone hepcidin in plasma or urine have not yet been widely available, whereas quantitative comparisons between hepcidin levels in these different matrices were thus far even impossible due to technical restrictions. To circumvent these limitations, we here describe several advances in time-of flight mass spectrometry (TOF MS), the most important of which concerned spiking of a synthetic hepcidin analogue as internal standard into serum and urine samples. This serves both as a control for experimental variation, such as recovery and matrix-dependent ionization and ion suppression, and at the same time allows value assignment to the measured hepcidin peak intensities. The assay improvements were clinically evaluated using samples from various patients groups and its relevance was further underscored by the significant correlation of serum hepcidin levels with serum iron indices in healthy individuals. Most importantly, this approach allowed kinetic studies as illustrated by the paired analyses of serum and urine samples, showing that more than 97% of the freely filtered serum hepcidin can be reabsorbed in the kidney. Thus, the here reported