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Bioanalysis ; 5(20): 2509-20, 2013 Oct.
Article in English | MEDLINE | ID: mdl-24138624

ABSTRACT

BACKGROUND: Hepcidin, a 25 amino acid peptide, plays an important role in iron homeostasis. Some hepcidin truncated peptides have antibiotic effects. RESULTS: A new analytical method for hepcidin determination in human plasma using LC-HRMS operating in full-scan acquisition mode has been validated. The extraction consists of protein precipitation and a drying reconstitution step; a 2.1 x 50 mm (idxL) C18 analytical column was used. Detection specificity, stability, accuracy, precision and recoveries were determined. The LOQ/LOD were 0.25/0.1 nM, respectively. More than 600 injections of plasma extracts were performed, allowing evaluation of the assay robustness. Hepcidin-20, hepcidin-22 and a new isoform, hepcidin-24, were detected in patients. CONCLUSION: The data underscore the usefulness of LC-HRMS for in-depth investigations related to hepcidin levels and pathways.


Subject(s)
Chromatography, Liquid/standards , Hepcidins/blood , Mass Spectrometry/standards , Amino Acid Sequence , Calibration , Female , Humans , Male , Mass Spectrometry/methods , Middle Aged , Molecular Sequence Data , Protein Isoforms/blood , Reproducibility of Results , Sensitivity and Specificity , Solid Phase Microextraction
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