Development of a standard reference material for vitamin D in
serum1– 4
Karen W Phinney
ABSTRACT
The most widely used indicator of vitamin D status is the mea-
surement 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. Investiga-
tors 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. Am J Clin Nutr 2008;
88(suppl):511S–2S.
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. Be-
cause circulating 25(OH)D can arise from hydroxylation of ei-
ther vitamin D2 or vitamin D3, measurement of total 25(OH)D
[both 25(OH)D2 and 25(OH)D3] is essential for accurate assess-
ment 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 par-
ticular cause for these discrepancies, some of the aspects of
25(OH)D measurements that researchers have considered in-
clude 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 ac-
curately the vitamin D status of individuals. In addition, the lack
of agreement in assay results obtained by different methods
Am J Clin Nutr 2008;88(suppl):511S–2S. Printed in USA. © 2008 American Society for Nutrition
might complicate the ability to define optimal levels of circulat-
ing 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
Downloaded from www.ajcn.org at West Virginia Univ on December 19, 2010
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 mea-
surements.
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 collab-
orating laboratories. NIST will perform its measurements by
isotope-dilution liquid chromatography-mass spectrometry and
tandem mass spectrometry methodology. The certificate of anal-
ysis 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 de-
signed the SRM to serve as a reproducible point of compari-
son. 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 devel-
oped and to assign values to in-house quality-control materi-
als. In addition, the new SRM can serve as an adjunct to
1
From the Analytical Chemistry Division, Chemical Science and Tech-
nology Laboratory, National Institute of Standards and Technology, Gaith-
ersburg, 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 iden-
tification 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@nist.gov.
511S
512S PHINNEY
existing quality assurance programs, such as the Vitamin D External
Quality Assessment Scheme, for vitamin D measurements.
The author had no conflicts of interest.
REFERENCES
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Downloaded from www.ajcn.org at West Virginia Univ on December 19, 2010