Simultaneous quantification of vitamin D3, 25-hydroxyvitamin D-3 and 24,25-dihydroxyvitamin D3 in human serum by LC-MS/MS

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Abstract

Introduction. Serum 25-hydroxy-vitamin D is the established biomarker of vitamin D status although serum concentrations of vitamin D and 24,25-dihydroxyvitamin D may also be of interest to understand the in vivo kinetics of serum 25-hydroxyvitamin D. Method. An LC-MS/MS method was developed and validated to quantify vitamin D-3, 25-hydroxyvitamin D-3 and 24,25-dihydroxyvitamin D-3 in serum. After protein precipitation of the serum it was loaded on a HybridSPE column to separate vitamin D metabolites from phospholipids. Vitamin D-3, 25-hydroxyvitamin D-3 and 24,25-dihydroxyvitamin D-3 in the eluate were derivatized by 4-phenyl-1,2,4-triazoline-3,5-dione to improve sensitivity in the following LC-MS/MS analysis. Results. Using only 100 L serum the limit of quantification was
Original languageEnglish
JournalScandinavian Journal of Clinical & Laboratory Investigation
Volume74
Issue number5
Pages (from-to)418-423
Number of pages6
ISSN0036-5513
DOIs
Publication statusPublished - 2014

Keywords

  • MEDICINE,
  • TANDEM MASS-SPECTROMETRY
  • LIQUID-CHROMATOGRAPHY
  • D METABOLITES
  • CIRCULATING VITAMIN-D-3
  • SUPPLEMENTATION
  • Liquid chromatography
  • vitamin D
  • mass spectroscopy

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