Mass spectrometry-based hepcidin measurements in serum and urine: analytical aspects and clinical implications.
Author(s): Kemna EH, Tjalsma H, Podust VN, Swinkels DW
Publication: Clin Chem, 2007, Vol. 53, Page 620-8
PubMed ID: 17272487 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
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Study Purpose
The purpose of this study was to determine the effects of freeze-thaw cycling, storage and circadian rhythms on hepcidin in urine and serum.
Summary of Findings:
Hepcidin-25 was detected in both serum and urine, but hepcidin-22 was only found in urine. Slight and inconsistent decreases were found in serum hepcidin levels after freeze-thaw cycling, but urine hepcidin consistently decreased after freeze-thaw cycling. The authors report that hepicidin in urine and serum was stable at -80 degrees C, but levels declined during 6 months of storage at -20 degrees C, even in the presence of PMSF. Serum levels of hepcidin peaked between 3 and 7 PM and were lowest between 3 and 6 AM, but urine levels of hepcidin showed only slight changes with circadian rhythms. Serum iron levels showed an inverse association with serum and urine hepcidin levels. Hepcidin concentrations were always lower when measured using the NP20 array compared to the IMAC30 array, and sensitivity was higher with the IMAC30 array. Concentrations of hepicidin in urine and serum were highest in patients with endotoxemia.
Biospecimens
Preservative Types
- Frozen
Diagnoses:
- Hemochromatosis
- Other diagnoses
Platform:
Analyte Technology Platform Peptide SELDI-TOF MS Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Storage Freeze/thaw cycling 1 cycle
2 cycles
3 cycles
4 cycles
Storage Storage temperature -20 degrees C
-80 degrees C
Storage Storage duration 0 months
6 months
Biospecimen Acquisition Time of biospecimen collection 6 AM
9 AM
12 PM
3 PM
6 PM
9 PM
12 AM
3 AM
SELDI-TOF MS Specific Type of array NP20
IMAC30
Analyte Extraction and Purification Protease inhibitor No protease inhibitor added
PMSF
Preaquisition Diagnosis/ patient condition Iron-deficiency anemia
Endotoxemia
Thalassemia major
Biospecimen Acquisition Biospecimen location Serum
Urine