The impact of delayed blood centrifuging, choice of collection tube, and type of assay on 25-hydroxyvitamin D concentrations

Cancer Causes Control. 2010 Apr;21(4):643-8. doi: 10.1007/s10552-009-9485-x. Epub 2009 Dec 18.


Studies have examined the associations between cancers and circulating 25-hydroxyvitamin D [25(OH)D], but little is known about the impact of different laboratory practices on 25(OH)D concentrations. We examined the potential impact of delayed blood centrifuging, choice of collection tube, and type of assay on 25(OH)D concentrations. Blood samples from 20 healthy volunteers underwent alternative laboratory procedures: four centrifuging times (2, 24, 72, and 96 h after blood draw); three types of collection tubes (red top serum tube, two different plasma anticoagulant tubes containing heparin or EDTA); and two types of assays (DiaSorin radioimmunoassay [RIA] and chemiluminescence immunoassay [CLIA/LIAISON((R))]). Log-transformed 25(OH)D concentrations were analyzed using the generalized estimating equations (GEE) linear regression models. We found no difference in 25(OH)D concentrations by centrifuging times or type of assay. There was some indication of a difference in 25(OH)D concentrations by tube type in CLIA/LIAISON((R))-assayed samples, with concentrations in heparinized plasma (geometric mean, 16.1 ng ml(-1)) higher than those in serum (geometric mean, 15.3 ng ml(-1)) (p = 0.01), but the difference was significant only after substantial centrifuging delays (96 h). Our study suggests no necessity for requiring immediate processing of blood samples after collection or for the choice of a tube type or assay.

Keywords: 25-hydroxyvitamin D; Epidemiologic methods; Specimen handling; Time factors; Vitamin D.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, N.I.H., Intramural

MeSH terms

  • Anticoagulants / administration & dosage
  • Blood Coagulation / drug effects
  • Blood Specimen Collection / instrumentation*
  • Blood Specimen Collection / methods*
  • Centrifugation
  • Clinical Laboratory Techniques / standards*
  • Edetic Acid / administration & dosage
  • Heparin / administration & dosage
  • Humans
  • Immunoassay / methods
  • Luminescent Measurements / methods
  • Quality Control
  • Radioimmunoassay / methods
  • Reproducibility of Results
  • Time Factors
  • Vitamin D / analogs & derivatives*
  • Vitamin D / blood


  • Anticoagulants
  • Vitamin D
  • Heparin
  • Edetic Acid
  • 25-hydroxyvitamin D