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

1. 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

   • Bodily Fluid - Serum
   • Bodily Fluid - Urine
   • Bodily Fluid - Blood

   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

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