Cancer Diagnosis Program | Biorepositories and Biospecimen Research Branch

Search BRD Papers

Hepcidin on 24-hour urine collection: preanalytical aspects and correlation with ferritin levels.

Author(s): Wolff F, Gentelet M, Gulbis B, Cotton F

Publication: Scand J Clin Lab Invest, 2016, Vol. 76, Page 454-9

PubMed ID: 27284811 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   This study investigated the effects of BSA addition, storage at room temperature and spot versus 24-h collection on the measurement of hepcidin in urine. The effect of 24-h collection versus spot collection on the correlation of urine hepcidin levels with plasma hepcidin and serum ferritin levels was also investigated. Blood and urine were collected from 33 healthy individuals, one patient on antibiotics for an ear infection, and a patient on quinolones for epididymitis. Blood was collected from each patient into polypropylene Venosafe serum, EDTA-K2, and lithium heparin tubes; spot urine was collected into containers with 0.065 g BSA, 0.125 g BSA or no BSA and 24 h urine into a container containing 5 g BSA. Levels of ferritin were measured immediately using blood from the serum tube. The lithium heparin blood was centrifuged at 4000 x g and plasma aliquots were stored at -80˚C in polypropylene tubes until hepcidin measurement by LC-MS/MS. Urine specimens were aliquoted and stored at -80˚C until analysis by LC-MS/MS.

   Summary of Findings:

   Spot urine with 2.5 g/L BSA had significantly higher hepcidin levels than spot urine without BSA (0.68 versus 0.59 nmol/mmol, P<0.001), but comparable levels to spot urine containing 5 g/L BSA (0.67 nmol/mmol). The calculated bias for collection without BSA was -13.3%. Hepcidin levels declined by 11.2% in spot urine specimens stored at room temperature for 24 h before aliquoting without BSA, but no decline was noted when specimens were stored with 2.5 g/L BSA. Similarly, hepcidin levels in 24 h urine were stable when specimens were stored at room temperature for 24 h prior to analysis. The authors report no effect of specimen pH on the measurement of hepcidin in urine. 

   Hepcidin levels were more strongly correlated between spot urine containing 2.5 g/L BSA and plasma (r=0.88) than between 24-h urine and plasma (r=0.78).  Although plasma hepcidin values were strongly correlated with serum ferritin levels (r=0.73), only modest correlations were observed for serum ferritin levels and hepcidin levels in 24 h urine (r=0.67) and spot urine with 2.5 g/L BSA (r=0.63). Finally, a cut-off of <0.9 nmol/L hepcidin in plasma had 100% sensitivity and 88.5% specificity for identifying iron deficiency.

   Biospecimens

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

   Preservative Types

   • Frozen

   Diagnoses:

   • Other diagnoses
   • Normal

   Platform:

   Analyte	Technology Platform
   Glycoprotein	Clinical chemistry/auto analyzer
   Electrolyte/Metal	Colorimetric assay
   Protein	LC-MS or LC-MS/MS
   Protein	Clinical chemistry/auto analyzer

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Acquisition	Biospecimen location	Plasma Urine Serum
   Biospecimen Aliquots and Components	Biospecimen components	0 g/L BSA 2.5 g/L BSA 5 g/L BSA
   Biospecimen Acquisition	Method of fluid acquisition	Voided urine (24-h collection) Voided urine (spot collection)

   View more