Department of Economics and Business Economics

A sensitive LC/MS/MS assay of 25OH vitamin D(3) and 25OH vitamin D(2) in dried blood spots

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

  • Darryl Eyles, Denmark
  • Cameron Anderson, Denmark
  • Pauline Ko, Denmark
  • Andrew Thomas, Australia
  • Alun Jones, Denmark
  • Thomas Burne, Denmark
  • Preben Bo Mortensen
  • Bent Nørgaard-Pedersen, Denmark
  • David Michael Hougaard, Denmark
  • John McGrath, Denmark
  • National Centre for Register-based Research
BACKGROUND: Low levels of 25 hydroxyvitamin D (25OHD) during early development is associated with a range of adverse health outcomes. While a number of methods exist to measure 25OHD in sera, none has been specifically developed to examine dried blood spots (DBS). Methods We describe an assay where 25 hydroxyvitamin D(3) (25OHD3) and 25hydroxyvitamin D(2) (25OHD2) are extracted from 3.2 mm DBS punches, derivatised with 4-phenyl-1,2,4-triazoline-3,5-dione (PTAD) prior to analysis with LC/MS/MS. We assessed assay precision, relative accuracy and examined the impact of storage conditions in samples stored for up to 22 years. RESULTS: The new assay had good accuracy and precision, and was highly sensitive, being capable of detecting <1 nmol/L 25OHD3 and 2 nmol/L 25OHD2. CDER sensitivity criteria were slightly higher at 7.7 nmol/L for 25OHD3 and 10.7 nmol/L for 25OHD2. The mean 25OHD3 concentration in 118 archived DBS was 20.8+/-11.4, (4.8 to 67.8 nmol/L). 25OHD2 was detected in only two of these samples. 25OHD3 concentrations were significantly higher in DBS collected in summer compared to winter (p<0.0001). CONCLUSION: Both 25OHD3 and 25OHD2 can be reliably quantified in archived 3.2 mm dried blood spots. We can not be certain that the levels we measure in archived samples are exactly the same as when they were collected. However, the fact that the DBS levels reflect the well-known seasonal variation in this vitamin and when corrected for sera, values fall within the normal range for 25OHD3, means that DBS are a useful tissue repository for testing a range of hypotheses linking developmental hypovitaminosis D and adverse health outcomes.
Original languageEnglish
JournalClinica Chimica Acta
Pages (from-to)145-151
Publication statusPublished - 2009

See relations at Aarhus University Citationformats

ID: 15343573