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American Journal of Clinical Nutrition, Vol. 88, No. 2, 511S-512S, August 2008
© 2008 American Society for Nutrition


Vitamin D and Health in the 21st Century: an Update

Development of a standard reference material for vitamin D in serum1,2,3,4

Karen W Phinney

1 From the Analytical Chemistry Division, Chemical Science and Technology Laboratory, National Institute of Standards and Technology, Gaithersburg, MD

2 Presented at the National Institutes of Health conference "Vitamin D and Health in the 21st Century: an Update," held in Bethesda, MD, September 5–6, 2007

3 Certain commercial equipment, instruments, or materials are identified in this report to specify adequately the experimental procedure. Such identification does not imply recommendation or endorsement by the National Institute of Standards and Technology, nor does it imply that the materials or equipment specified are necessarily the best available for the purpose.

4 Address reprint requests to KW Phinney, National Institute of Standards and Technology, 100 Bureau Drive, Stop 8392, Gaithersburg, MD 20899-8392. E-mail: karen.phinney{at}nist.gov.

ABSTRACT

The most widely used indicator of vitamin D status is the measurement of 25-hydroxyvitamin D [25(OH)D] in either serum or plasma. Several studies have reported discrepancies between the results of assays used to measure 25(OH)D, however, which calls into question the ability of 25(OH)D assays to reflect accurately the vitamin D status of individuals. The National Institute of Standards and Technology has been working with the National Institutes of Health's Office of Dietary Supplements to develop a standard reference material for circulating vitamin D analysis. This standard reference material will provide a material with stable, well-defined levels of the analytes of interest. Investigators will be able to use the standard reference material to validate new analytic methods as they are developed and to assign values to in-house quality-control materials.

STANDARD REFERENCE MATERIAL FOR VITAMIN D

The prevalence of vitamin D deficiency or insufficiency in the general population is an issue of concern. The most widely used indicator of vitamin D status is the measurement of 25-hydroxyvitamin D [25(OH)D] in either serum or plasma. Because circulating 25(OH)D can arise from hydroxylation of either vitamin D2 or vitamin D3, measurement of total 25(OH)D [both 25(OH)D2 and 25(OH)D3] is essential for accurate assessment of vitamin D status (1).

Several studies, including interlaboratory comparisons, have reported discrepancies between the results of assays used to measure 25(OH)D (2, 3). Data from the Vitamin D External Quality Assessment Scheme have also illustrated assay-specific results (4). Although it is difficult to point to a particular cause for these discrepancies, some of the aspects of 25(OH)D measurements that researchers have considered include the assays' ability to respond equally to 25(OH)D2 and 25(OH)D3, preparation of calibrants, and the recovery of 25(OH)D from spiked samples (5). In addition, a recently discovered metabolite, 3-epi-25-hydroxyvitamin D [3-epi-25(OH)D], which might be present in samples from infants, can pose problems for certain assays (6).

The current level of variability in 25(OH)D measurements calls into question the ability of 25(OH)D assays to reflect accurately the vitamin D status of individuals. In addition, the lack of agreement in assay results obtained by different methods might complicate the ability to define optimal levels of circulating 25(OH)D (3). For these reasons, investigators have called for international standardization of vitamin D measurements.

The National Institute of Standards and Technology (NIST) has been working with the National Institutes of Health's Office of Dietary Supplements to develop a standard reference material (SRM) for circulating vitamin D analysis. NIST has a long history of providing SRMs for the clinical chemistry community to support accuracy in clinical laboratory measurements.

The reference material currently in development at NIST, SRM 972 Vitamin D in Human Serum, consists of 4 pools of fresh-frozen serum. Each pool has a different level of 25(OH)D2, 25(OH)D3, or both. One pool also contains 3-epi-25(OH)D3. NIST designed the SRM to pose similar analytic challenges to those encountered in patient samples. NIST will assign values for each of the analytes through measurements at NIST and collaborating laboratories. NIST will perform its measurements by isotope-dilution liquid chromatography-mass spectrometry and tandem mass spectrometry methodology. The certificate of analysis for SRM 972 will include values for 25(OH)D2, 25(OH)D3, and 3-epi-25(OH)D3. NIST anticipates official release of SRM 972 in 2008.

This new SRM is not intended to characterize any particular analytic method as superior to another. Instead, NIST designed the SRM to serve as a reproducible point of comparison. The SRM will provide a material with stable, well-defined levels of the analytes of interest. Investigators can use the SRM to validate new analytic methods as they are developed and to assign values to in-house quality-control materials. In addition, the new SRM can serve as an adjunct to existing quality assurance programs, such as the Vitamin D External Quality Assessment Scheme, for vitamin D measurements.

ACKNOWLEDGMENTS

The author had no conflicts of interest.

REFERENCES

  1. Hollis B. The determination of circulating 25-hydroxyvitamin D: no easy task. J Clin Endocrinol Metab 2004; 89: 3149–51.[Free Full Text]
  2. Glendenning P, Noble J, Taranto M, et al. Issues of methodology, standardization and metabolite recognition for 25-hydroxyvitamin D when comparing the DiaSorin radioimmunoassay and the Nichols Advantage automated chemiluminescence protein-binding assay in hip fracture cases. Ann Clin Biochem 2003; 40: 546–51.[Abstract/Free Full Text]
  3. Binkley N, Krueger D, Cowgill C, et al. Assay variation confounds the diagnosis of hypovitaminosis D: a call for standardization. J Clin Endocrinol Metab 2004; 89: 3152–7.[Abstract/Free Full Text]
  4. Carter G, Carter R, Jones J, Berry J. How accurate are assays for 25-hydroxyvitamin D? Data from the International Vitamin D External Quality Assessment Scheme. Clin Chem 2004; 50: 2195–7.
  5. Lensmeyer G, Wiebe D, Binkley N, Drezner M. HPLC method for 25-hydroxyvitamin D measurement: comparison with contemporary assays. Clin Chem 2006; 52: 1120–6.[Abstract/Free Full Text]
  6. Singh R, Taylor R, Reddy G, Grebe S. C-3 epimers can account for a significant proportion of total circulating 25-hydroxyvitamin D in infants, complicating accurate measurement and interpretation of vitamin D status. J Clin Endocrinol Metab 2006; 91: 3055–61.[Abstract/Free Full Text]



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