2. Academic Validation
  3. 1β,25-Dihydroxyvitamin D3: A new vitamin D metabolite in human serum

1β,25-Dihydroxyvitamin D3: A new vitamin D metabolite in human serum

  • J Steroid Biochem Mol Biol. 2017 Oct;173:341-348. doi: 10.1016/j.jsbmb.2017.02.004.
Steven Pauwels 1,  Ivo Jans 2,  Jaak Billen 3,  Annemieke Heijboer 4,  Annemieke Verstuyf 5,  Geert Carmeliet 5,  Chantal Mathieu 5,  Miguel Maestro 6,  Etienne Waelkens 7,  Pieter Evenepoel 8,  Roger Bouillon 9,  Dirk Vanderschueren 3,  Pieter Vermeersch 1
Affiliations

Affiliations

  • 1 Department of Laboratory Medicine, University Hospitals Leuven, 3000 Leuven, Belgium; Department of Cardiovascular Sciences, KU Leuven, 3000 Leuven, Belgium.
  • 2 Department of Laboratory Medicine, University Hospitals Leuven, 3000 Leuven, Belgium.
  • 3 Department of Laboratory Medicine, University Hospitals Leuven, 3000 Leuven, Belgium; Laboratory of Clinical and Experimental Endocrinology, KU Leuven, 3000 Leuven, Belgium.
  • 4 Endocrine Laboratory, Department of Clinical Chemistry, VU University medical center, Amsterdam, the Netherlands.
  • 5 Laboratory of Clinical and Experimental Endocrinology, KU Leuven, 3000 Leuven, Belgium.
  • 6 Department of Fundamental Chemistry, University of A Coruña, A Coruña, Spain.
  • 7 Department of Cellular and Molecular Medicine, KU Leuven, 3000 Leuven, Belgium.
  • 8 Department of Immunology and Microbiology, Laboratory of Nephrology, KU Leuven, 3000 Leuven, Belgium.
  • 9 Laboratory of Clinical and Experimental Endocrinology, KU Leuven, 3000 Leuven, Belgium. Electronic address: [email protected].
PMID: 28193495 DOI: 10.1016/j.jsbmb.2017.02.004
Abstract

Background: The measurement of 1α,25(OH)2D3 in human serum poses a true challenge as concentrations are very low and structurally similar metabolites can interfere.

Materials and methods: During optimization of our in-house LC-MSMS method for serum 1α,25(OH)2D3 a previously co-eluting isobaric interference was separated. The isobar was identified as 1β,25(OH)2D3 by comparing retention time and fragmentation spectra to standards (other isobaric dihydroxylated vitamin D3 analogs). 1β,25(OH)2D3 showed specific cluster formation (water), not present in 1α,25(OH)2D3. 1β,25(OH)2D3 was measured in serum of apparently healthy human volunteers (n=20), patients with high serum 25-hydroxyvitamin D [25(OH)D] concentrations (>50ng/mL) (n=33 among which 4 with very high levels (>150ng/mL)) and patients with kidney failure (n=68; 39 stage 1-3, 29 stage 4-5). Pearson's r was calculated for correlations and Mann-Whitney statistic to compare group medians.

Results: Median serum 1β,25(OH)2D3 was 11pg/mL in apparently healthy volunteers and increased to 20pg/mL for serum 25(OH)D concentrations above 80ng/mL (n=22) (p<0.0001). 1β,25(OH)2D3 concentrations were significantly correlated to serum 25(OH)D concentrations (r=0.85) for the combined results from healthy volunteers and patient sera (n=53) (p<0.0001). For patients with kidney failure, median serum 1β,25(OH)2D3 was 7pg/mL and not different from the median level in healthy volunteers (p=0.06). The median concentration did not vary with different stages.

Conclusions: We present evidence for the widespread presence of 1β,25(OH)2D3, a new vitamin D metabolite, in human serum. The level increases with rising serum 25(OH)D concentrations and is particularly high in patients with very high 25(OH)D levels. We previously demonstrated that 1β,25(OH)2D3 is a poor genomic agonist but a potent non-genomic antagonist of 1α,25(OH)2D3. The clinical implications of the presence of this analog therefore require further exploration.

Keywords

1,25-dihydroxyvitamin D; Liquid chromatography; Mass Spectrometry; Metabolism; Vitamin D.

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