Aim: Hepcidin-25 is the master regulator of iron homeostasis. N-truncated isoforms of hepcidin-25 have been identified (hepcidin-20, -22, -24), although data on the concentrations of these isoforms are sparse.
Materials & methods: Serum was mixed with aqueous formic acid, and the supernatant loaded onto a 96-well-SPE-plate. Eluted analytes were analyzed using LC-HR-MS. Forty-seven paired dipotassium-EDTA human plasma and serum samples were analyzed.
Results: The LLOQ was 1 μg/l (all analytes). Accuracy and precision were acceptable. There was a good correlation (R2 >0.90, all analytes) between matrices. The median (range) serum hepcidin-20, -22, -24 and -25 concentrations measured were 4 (1-40), 8 (2-20), 8 (1-50) and 39 (1-334) μg/l, respectively.
Conclusion: LC-HR-MS is widely applicable to the measurement of hepcidin-25, and truncated isoforms.
Keywords: hepcidin-25; isoforms; plasma; serum.