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Cited by 140Vitamin D Research StudiesClinical Laboratory Practices and Quality ControlMetabolomics and Mass Spectrometry Studies

Measurement by a Novel LC-MS/MS Methodology Reveals Similar Serum Concentrations of Vitamin D–Binding Protein in Blacks and Whites

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Henderson, Clark M, Lutsey, Pamela L, Misialek, Jeffrey R, Laha, Thomas J, Selvin, Elizabeth, Eckfeldt, John H, Hoofnagle, Andrew N·2016·Clinical Chemistry
DOI 10.1373/clinchem.2015.244541PMC PMC4698095
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Abstract

BACKGROUND: Vitamin D deficiency is associated with poor bone health and other adverse health outcomes; however, the associations are greatly attenuated in black vs white individuals. One possible explanation for this attenuation is different concentrations of bioavailable vitamin D metabolites in plasma, which are estimated with equations that include the total concentration of vitamin D binding globulin (VDBG) and haplotype-specific dissociation constants. METHODS: We developed a method to quantify VDBG with LC-MS/MS that could also identify the haplotypes/isoforms of VDBG present. We validated the method according to recent recommendations for publications of biomarker studies. We determined serum VDBG concentrations in samples from the Atherosclerosis Risk in Communities cohort and compared the results with a widely used monoclonal immunoassay. RESULTS: With 10 μL of serum or plasma, the lower limit of quantification for the assay (<20% CV) was 71 μg/mL. The assay was linear from 62 to 434 μg/mL, with total imprecision of 7.3–9.0% CV at approximately 250 μg/mL. Significant hemolysis interfered with quantification. The identification of isoforms was 97% concordant with genotyping (κ coefficient). Method comparison with immunoassay revealed significant isoform-specific effects in the immunoassay. Mean concentrations (SD) of VDBG by mass spectrometry were similar in whites and blacks [262 (25) vs 266 (35) μg/mL, respectively; P = 0.43]. CONCLUSIONS: Validated mass spectrometric methods for the quantification of proteins in human samples can provide additional information beyond immunoassay. Counter to prior observations by immunoassay, VDBG concentrations did not vary by race.

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  • No racial difference in DBP was observed when using a polyclonal assay or applying LC-MS/MS, so the earlier conclusion was most likely the result of a monoclonal assay bias, confirming poor vitamin D status in subjects of African origin.

    “However, as no racial difference in DBP was observed when using a polyclonal assay or applying LC–MS/MS, this conclusion was most likely the result of a monoclonal assay bias, confirming poor vitamin D status in subjects of African origin [81, 82].”

    From Role of sex hormone-binding globulin in the free hormone hypothesis and the relevance of free testosterone in androgen physiology (2022) · cites this paper as [82]

    The Results (Racial distributions; page unavailable) report no racial difference in vitamin D–binding protein measured by LC-MS/MS, despite a difference measured by the immunoassay. The Discussion (Estimates of free and bioavailable vitamin D metabolites; page unavailable) says the monoclonal assay may yield misleading results and reports that 81% of Black people fall below a standard 25(OH)D cutoff. This paper does not report a polyclonal assay finding of no racial difference, and it cautions that the cutoff may not fully capture vitamin D sufficiency; it therefore does not confirm poor overall vitamin D status in people of African origin. (LLM, conf=0.86)

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Authors
Henderson, C. M., Lutsey, P. L., Misialek, J. R., Laha, T. J., Selvin, E., Eckfeldt, J. H., & Hoofnagle, A. N.
Affiliations
  • Department of Laboratory Medicine and
  • Division of Epidemiology and Community Health
  • Johns Hopkins University (US)
  • University of Minnesota (US)
  • University of Washington (US)
Corresponding author
Hoofnagle, Andrew N
ORCID
Venue
Clinical Chemistry
Publisher
Oxford University Press (OUP)
Published
2016-01-01
Volume
62
Issue
1
Pages
179-187
ISSN
  • 0009-9147 (ISSN-L)
  • 1530-8561
DOI
10.1373/clinchem.2015.244541
PMC
PMC4698095
OpenAlex
W2178622146
Cited by
140
References
1
Citations by year
2026: 4 · 2025: 5 · 2024: 9 · 2023: 9 · 2022: 12 · 2021: 11 · 2020: 16 · 2019: 11 · 2018: 17 · 2017: 27 · 2016: 15 · 2015: 3
FWCI
16.42
Type
journal article
Language
en
Open access
Licence
academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model (vor)
Locations
  • Clinical Chemistry (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • National Heart, Lung, and Blood Institute (HHSN268201100012C, HHSN268201100011C, HHSN268201100010C, HHSN268201100009C, HHSN268201100008C, HHSN268201100007C, HHSN268201100006C, HHSN268201100005C)
  • University of Washington
  • NIH (DK035816, CA160034, DK089174, HL103706-S1, HL103706, HL007028)
Topics
  • Health Sciences › Medicine › Pathology and Forensic Medicine › Vitamin D Research Studies
  • Health Sciences › Medicine › Physiology › Clinical Laboratory Practices and Quality Control
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Metabolomics and Mass Spectrometry Studies
Keywords
vitamin D binding protein, vitamin D deficiency, serum protein quantification, LC-MS/MS method validation, racial differences, Atherosclerosis Risk in Communities Study, hemolysis interference
MeSH terms
Black or African American, Chromatography, Liquid, Female, Genotype, Humans, Immunoassay, Male, United States, Vitamin D-Binding Protein, Black People, White People, Tandem Mass Spectrometry

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