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Determination of human reference values for serum total 1,25-dihydroxyvitamin D using an extensively validated 2D
Niek F Dirks1, Frans Martens1, Dirk Vanderschueren2
1Department of Clinical Chemistry, Endocrine Laboratory, VU University Medical Center, Amsterdam, The Netherlands.
The Journal of Steroid Biochemistry and Molecular Biology
|December 23, 2015
Summary
This study establishes a reference interval of 59-159pmol/L for 1,25-dihydroxyvitamin D3 using a validated LC-MS/MS method. This provides crucial data for assessing vitamin D status and diagnosing related disorders.
Area of Science:
- Biochemistry
- Endocrinology
- Clinical Chemistry
Background:
- Assessing vitamin D status typically involves measuring 25-hydroxyvitamin D.
- Measurement of 1,25-dihydroxyvitamin D is critical for diagnosing specific vitamin D metabolism disorders, including 1a-hydroxylation issues and vitamin D receptor defects.
Purpose of the Study:
- To establish reference values for 1,25-dihydroxyvitamin D3 and D2.
- To validate a 2D ID-UPLC-MS/MS method for precise measurement of these metabolites.
Main Methods:
- A 2D ID-UPLC-MS/MS method was developed and validated for quantifying 1,25-dihydroxyvitamin D3 and D2 in human serum.
- The method demonstrated high precision (intra-assay <5-11%) and a low limit of quantitation (3.4 pmol/L).
- Correlation analysis showed good agreement with existing LC-MS/MS methods and external quality assessment schemes.
Main Results:
- The validated LC-MS/MS method accurately measured picomolar concentrations of 1,25-dihydroxyvitamin D3 and D2.
- Established reference ranges in healthy volunteers: 59-159 pmol/L for 1,25-dihydroxyvitamin D3 and <17 pmol/L for 1,25-dihydroxyvitamin D2.
- Observed age-related decrease in 1,25-dihydroxyvitamin D3 in women and higher levels in premenopausal women using oral contraceptives, suggesting an estrogen influence.
Conclusions:
- The study successfully determined a precise reference interval for 1,25-dihydroxyvitamin D3 (59-159 pmol/L) using an advanced LC-MS/MS technique.
- These findings contribute essential reference data for clinical diagnostics and research related to vitamin D metabolism.
