Interlaboratory comparison of 25-hydroxyvitamin D assays: Vitamin D Standardization Program (VDSP) Intercomparison Study 2 - Part 2 ligand binding assays - impact of 25-hydroxyvitamin D2 and 24R,25-dihydroxyvitamin D3 on assay performance

doi:10.1007/s00216-021-03577-0

. 2022 Jan;414(1):351-366.

doi: 10.1007/s00216-021-03577-0. Epub 2021 Aug 25.

Interlaboratory comparison of 25-hydroxyvitamin D assays: Vitamin D Standardization Program (VDSP) Intercomparison Study 2 - Part 2 ligand binding assays - impact of 25-hydroxyvitamin D₂ and 24R,25-dihydroxyvitamin D₃ on assay performance

Stephen A Wise^{1

2}, Johanna E Camara³, Carolyn Q Burdette³, Grace Hahm³, Federica Nalin³, Adam J Kuszak⁴, Joyce Merkel⁴, Ramón A Durazo-Arvizu⁵, Emma L Williams⁶, Christian Popp⁷, Christian Beckert⁷, Jan Schultess⁷, Glen Van Slooten⁸, Carole Tourneur⁹, Camille Pease⁹, Ravi Kaul¹⁰, Alfredo Villarreal¹⁰, Marcelo Cidade Batista¹¹, Heather Pham¹², Alex Bennett¹², Eugene Jansen¹³, Dilshad Ahmed Khan¹⁴, Mark Kilbane¹⁵, Patrick J Twomey¹⁵, James Freeman¹⁶, Neil Parker¹⁶, Sohail Mushtaq¹⁷, Christine Simpson¹⁸, Pierre Lukas¹⁹, Étienne Cavalier¹⁹, Christopher T Sempos^{4

20}

Affiliations

¹ Office of Dietary Supplements (ODS), National Institutes of Health (NIH), Bethesda, MD, 20892, USA. stephen.wise@nih.gov.
² Scientist Emeritus, National Institute of Standards and Technology (NIST), MD, 20899, Gaithersburg, USA. stephen.wise@nih.gov.
³ Chemical Sciences Division, National Institute of Standards and Technology (NIST), Gaithersburg, MD, 20899, USA.
⁴ Office of Dietary Supplements (ODS), National Institutes of Health (NIH), Bethesda, MD, 20892, USA.
⁵ Biostatistics Core, The Sabin Research Institute, Keck School of Medicine, University of Southern California, Los Angeles, CA, 90027, USA.
⁶ Imperial Healthcare NHS Trust, London, W6 8RF, UK.
⁷ Abbott Laboratories, ADD Wiesbaden Abbott GmbH, 65205, Wiesbaden, Germany.
⁸ Awareness Technology, Inc., Palm City, FL, 34990, USA.
⁹ bioMérieux, 69280, Marcy-l'Étoile, France.
¹⁰ Clinical Diagnostics, Bio-Rad Laboratories, Hercules, CA, 94547, USA.
¹¹ Clinical Lab, Hospital Israelita Albert Einstein, São Paulo, 05652-900, Brazil.
¹² Immunodiagnostic Systems (IDS), Boldon, NE35 9PD, UK.
¹³ National Institute of Public Health and the Environment (RIVM), 3720, Bilthoven, BA, Netherlands.
¹⁴ National University of Medical Sciences (NUMS), Rawalpindi, 46000, Pakistan.
¹⁵ Clinical Chemistry, St. Vincent's University Hospital, Elm Park, Dublin 4, D04 T6F4, Ireland.
¹⁶ Siemens-Healthineers, Tarrytown, NY, 10591, USA.
¹⁷ University of Chester, Chester, CH1 4BJ, UK.
¹⁸ School of Medicine, Yale University, New Haven, CT, USA.
¹⁹ Clinical Chemistry, University of Liège, CHU de Liège, 4000, Liège, Belgium.
²⁰ Vitamin D Standardization Program LLC, Havre de Grace, MD, 21078, USA.

PMID: 34435207
PMCID: PMC8748290
DOI: 10.1007/s00216-021-03577-0

Interlaboratory comparison of 25-hydroxyvitamin D assays: Vitamin D Standardization Program (VDSP) Intercomparison Study 2 - Part 2 ligand binding assays - impact of 25-hydroxyvitamin D₂ and 24R,25-dihydroxyvitamin D₃ on assay performance

Stephen A Wise et al. Anal Bioanal Chem. 2022 Jan.

. 2022 Jan;414(1):351-366.

doi: 10.1007/s00216-021-03577-0. Epub 2021 Aug 25.

Authors

Affiliations

¹ Office of Dietary Supplements (ODS), National Institutes of Health (NIH), Bethesda, MD, 20892, USA. stephen.wise@nih.gov.
² Scientist Emeritus, National Institute of Standards and Technology (NIST), MD, 20899, Gaithersburg, USA. stephen.wise@nih.gov.
³ Chemical Sciences Division, National Institute of Standards and Technology (NIST), Gaithersburg, MD, 20899, USA.
⁴ Office of Dietary Supplements (ODS), National Institutes of Health (NIH), Bethesda, MD, 20892, USA.
⁵ Biostatistics Core, The Sabin Research Institute, Keck School of Medicine, University of Southern California, Los Angeles, CA, 90027, USA.
⁶ Imperial Healthcare NHS Trust, London, W6 8RF, UK.
⁷ Abbott Laboratories, ADD Wiesbaden Abbott GmbH, 65205, Wiesbaden, Germany.
⁸ Awareness Technology, Inc., Palm City, FL, 34990, USA.
⁹ bioMérieux, 69280, Marcy-l'Étoile, France.
¹⁰ Clinical Diagnostics, Bio-Rad Laboratories, Hercules, CA, 94547, USA.
¹¹ Clinical Lab, Hospital Israelita Albert Einstein, São Paulo, 05652-900, Brazil.
¹² Immunodiagnostic Systems (IDS), Boldon, NE35 9PD, UK.
¹³ National Institute of Public Health and the Environment (RIVM), 3720, Bilthoven, BA, Netherlands.
¹⁴ National University of Medical Sciences (NUMS), Rawalpindi, 46000, Pakistan.
¹⁵ Clinical Chemistry, St. Vincent's University Hospital, Elm Park, Dublin 4, D04 T6F4, Ireland.
¹⁶ Siemens-Healthineers, Tarrytown, NY, 10591, USA.
¹⁷ University of Chester, Chester, CH1 4BJ, UK.
¹⁸ School of Medicine, Yale University, New Haven, CT, USA.
¹⁹ Clinical Chemistry, University of Liège, CHU de Liège, 4000, Liège, Belgium.
²⁰ Vitamin D Standardization Program LLC, Havre de Grace, MD, 21078, USA.

PMID: 34435207
PMCID: PMC8748290
DOI: 10.1007/s00216-021-03577-0

Abstract

An interlaboratory comparison study was conducted by the Vitamin D Standardization Program (VDSP) to assess the performance of ligand binding assays (Part 2) for the determination of serum total 25-hydroxyvitamin D [25(OH)D]. Fifty single-donor samples were assigned target values for concentrations of 25-hydroxyvitamin D₂ [25(OH)D₂], 25-hydroxyvitamin D₃ [25(OH)D₃], 3-epi-25-hydroxyvitamin D₃ [3-epi-25(OH)D₃], and 24R,25-dihydroxyvitamin D₃ [24R,25(OH)₂D₃] using isotope dilution liquid chromatography-tandem mass spectrometry (ID LC-MS/MS). VDSP Intercomparison Study 2 Part 2 includes results from 17 laboratories using 32 ligand binding assays. Assay performance was evaluated using mean % bias compared to the assigned target values and using linear regression analysis of the test assay mean results and the target values. Only 50% of the ligand binding assays achieved the VDSP criterion of mean % bias ≤ |± 5%|. For the 13 unique ligand binding assays evaluated in this study, only 4 assays were consistently within ± 5% mean bias and 4 assays were consistently outside ± 5% mean bias regardless of the laboratory performing the assay. Based on multivariable regression analysis using the concentrations of individual vitamin D metabolites in the 50 single-donor samples, most assays underestimate 25(OH)D₂ and several assays (Abbott, bioMérieux, DiaSorin, IDS-EIA, and IDS-iSYS) may have cross-reactivity from 24R,25(OH)₂D₃. The results of this interlaboratory study represent the most comprehensive comparison of 25(OH)D ligand binding assays published to date and is the only study to assess the impact of 24R,25(OH)₂D₃ content using results from a reference measurement procedure.

Keywords: 24R,25-Dihydroxyvitamin D3 [24R,25(OH)2D3]; 25-Hydroxyvitamin D2 [25(OH)D2]; 25-Hydroxyvitamin D3 [25(OH)D3]; Ligand binding assay; Liquid chromatography–tandem mass spectrometry (LC-MS/MS); Total 25-hydroxyvitamin D [25(OH)D].

PubMed Disclaimer

Conflict of interest statement

Conflict of Interest

S.A. Wise is an Editor of the journal Analytical and Bioanalytical Chemistry and was not involved in peer reviewing this manuscript. Several of the coauthors are employees of companies that produce assays that were evaluated in this study. There are no financial or nonfinancial competing interests for any of the coauthors.

Figures

**Figure 1.**
Results for determination of serum total 25(OH)D in single-donor samples versus the NIST assigned target value for the DiaSorin assay (Lab 9) (A and B) and Abbott assay (Lab 1) (C and D).

**Figure 2.**
Results for determination of serum total 25(OH)D in single-donor samples versus the NIST assigned target value for the Siemens assay (Lab 30) (A and B) and Bio-Rad assay (Lab 4) (C and D).

**Figure 3.**
Mean % bias for the determination of serum total 25(OH)D in 50 single-donor samples compared with the NIST target values for Abbott assay (Lab 1) (A), DiaSorin assay (Lab 9) (B), IDS-iSYS-2 assay (Lab 20) (C), and bioMérieux assay (Lab 3) (D).

**Figure 4.**
Mean % bias for various ligand binding assays for single-donor samples with normal concentrations of 25(OH)D₂ (42 samples) (yellow bar) and with high concentrations (>30 nmol/L) of 25(OH)D₂ (8 samples) (green bar). Error bars are the SD of the % mean bias for the sample sets.

See this image and copyright information in PMC

Cited by

Towards harmonization of directly measured free 25-hydroxyvitamin D using an enzyme-linked immunosorbent assay.
Sempos CT, Lindhout E, Heureux N, Hars M, Parkington DA, Dennison E, Durazo-Arvizu R, Jones KS, Wise SA. Sempos CT, et al. Anal Bioanal Chem. 2022 Nov;414(27):7793-7803. doi: 10.1007/s00216-022-04313-y. Epub 2022 Sep 16. Anal Bioanal Chem. 2022. PMID: 36109397 Free PMC article.
Multicenter comparison of analytical interferences of 25-OH vitamin D immunoassay and mass spectrometry methods by endogenous interferents and cross-reactivity with 3-epi-25-OH-vitamin D₃.
Lee JH, Seo JD, Lee K, Roh EY, Yun YM, Lee YW, Cho SE, Song J. Lee JH, et al. Pract Lab Med. 2023 Dec 12;38:e00347. doi: 10.1016/j.plabm.2023.e00347. eCollection 2024 Jan. Pract Lab Med. 2023. PMID: 38188654 Free PMC article.
Determination of optimal 25-hydroxyvitamin D cutoff values for the evaluation of restless legs syndrome among pregnant women.
Miyazaki A, Takahashi M, Shuo T, Eto H, Kondo H. Miyazaki A, et al. J Clin Sleep Med. 2023 Jan 1;19(1):73-83. doi: 10.5664/jcsm.10270. J Clin Sleep Med. 2023. PMID: 35999809 Free PMC article.
Discordant Health Implications and Molecular Mechanisms of Vitamin D in Clinical and Preclinical Studies of Prostate Cancer: A Critical Appraisal of the Literature Data.
Fendler A, Stephan C, Ralla B, Jung K. Fendler A, et al. Int J Mol Sci. 2024 May 13;25(10):5286. doi: 10.3390/ijms25105286. Int J Mol Sci. 2024. PMID: 38791324 Free PMC article. Review.
A candidate reference method and multiple commutable control materials for serum 25-hydroxyvitamin D measurement.
Zhang L, Long Q, Zhang J, Zeng Q, Zhao H, Chen W, Zhang T, Zhang C. Zhang L, et al. J Clin Lab Anal. 2022 Dec;36(12):e24756. doi: 10.1002/jcla.24756. Epub 2022 Nov 13. J Clin Lab Anal. 2022. PMID: 36371780 Free PMC article.

See all "Cited by" articles

References

1. Sempos CT, Vesper HW, Phinney KW, Thienpont LM, Coates PM, VDSP (2012) Vitamin D status as an international issue: National surveys and the problem of standardization. Scand J Clin Lab Invest 72:32–40. doi:10.3109/00365513.2012.681935 - DOI - PubMed
1. Binkley N, Dawson-Hughes B, Durazo-Arvizu R, Thamm M, Tian L, Merkel JM, et al. (2017) Vitamin D measurement standardization: The way out of the chaos. J Steroid Biochem Mol Biol 173:117–121. doi:10.1016/j.jsbmb.2016.12.002 - DOI - PubMed
1. Binkley N, Krueger D, Cowgill CS, Plum L, Lake E, Hansen KE, et al. (2004) Assay variation confounds the diagnosis of hypovitaminosis D: A call for standardization. J Clin Endocrinol Metab 89 (7):3152–3157. doi:10.1210/jc.2003-031979 - DOI - PubMed
1. Wise SA, Phinney KW, Tai SSC, Camara JE, Myers GL, Durazo-Arvizu R, et al. (2017) Baseline assessment of 25-hydroxyvitamin D assay performance: A Vitamin D Standardization Program (VDSP) interlaboratory comparison study. J AOAC Int 100 (5):1244–1252. doi:10.5740/jaoacint.17-0258 - DOI - PubMed
1. Sempos CT, Binkley N (2020) 25-Hydroxvitamin D assay standardization and vitamin D guidelines paralysis. Public Health Nutrition 23 (7):1153–1164. doi:10.1017/S1368980019005251 - DOI - PMC - PubMed

MeSH terms

Substances

Grants and funding

LinkOut - more resources

Full Text Sources
Medical
- MedlinePlus Health Information

[1] Sempos CT, Vesper HW, Phinney KW, Thienpont LM, Coates PM, VDSP (2012) Vitamin D status as an international issue: National surveys and the problem of standardization. Scand J Clin Lab Invest 72:32–40. doi:10.3109/00365513.2012.681935 - DOI - PubMed

[2] Sempos CT, Vesper HW, Phinney KW, Thienpont LM, Coates PM, VDSP (2012) Vitamin D status as an international issue: National surveys and the problem of standardization. Scand J Clin Lab Invest 72:32–40. doi:10.3109/00365513.2012.681935 - DOI - PubMed

[3] Binkley N, Dawson-Hughes B, Durazo-Arvizu R, Thamm M, Tian L, Merkel JM, et al. (2017) Vitamin D measurement standardization: The way out of the chaos. J Steroid Biochem Mol Biol 173:117–121. doi:10.1016/j.jsbmb.2016.12.002 - DOI - PubMed

[4] Binkley N, Dawson-Hughes B, Durazo-Arvizu R, Thamm M, Tian L, Merkel JM, et al. (2017) Vitamin D measurement standardization: The way out of the chaos. J Steroid Biochem Mol Biol 173:117–121. doi:10.1016/j.jsbmb.2016.12.002 - DOI - PubMed

[5] Binkley N, Krueger D, Cowgill CS, Plum L, Lake E, Hansen KE, et al. (2004) Assay variation confounds the diagnosis of hypovitaminosis D: A call for standardization. J Clin Endocrinol Metab 89 (7):3152–3157. doi:10.1210/jc.2003-031979 - DOI - PubMed

[6] Binkley N, Krueger D, Cowgill CS, Plum L, Lake E, Hansen KE, et al. (2004) Assay variation confounds the diagnosis of hypovitaminosis D: A call for standardization. J Clin Endocrinol Metab 89 (7):3152–3157. doi:10.1210/jc.2003-031979 - DOI - PubMed

[7] Wise SA, Phinney KW, Tai SSC, Camara JE, Myers GL, Durazo-Arvizu R, et al. (2017) Baseline assessment of 25-hydroxyvitamin D assay performance: A Vitamin D Standardization Program (VDSP) interlaboratory comparison study. J AOAC Int 100 (5):1244–1252. doi:10.5740/jaoacint.17-0258 - DOI - PubMed

[8] Wise SA, Phinney KW, Tai SSC, Camara JE, Myers GL, Durazo-Arvizu R, et al. (2017) Baseline assessment of 25-hydroxyvitamin D assay performance: A Vitamin D Standardization Program (VDSP) interlaboratory comparison study. J AOAC Int 100 (5):1244–1252. doi:10.5740/jaoacint.17-0258 - DOI - PubMed

[9] Sempos CT, Binkley N (2020) 25-Hydroxvitamin D assay standardization and vitamin D guidelines paralysis. Public Health Nutrition 23 (7):1153–1164. doi:10.1017/S1368980019005251 - DOI - PMC - PubMed

[10] Sempos CT, Binkley N (2020) 25-Hydroxvitamin D assay standardization and vitamin D guidelines paralysis. Public Health Nutrition 23 (7):1153–1164. doi:10.1017/S1368980019005251 - DOI - PMC - PubMed

Save citation to file

Email citation

Add to Collections

Add to My Bibliography

Your saved search

Create a file for external citation management software

Your RSS Feed

Interlaboratory comparison of 25-hydroxyvitamin D assays: Vitamin D Standardization Program (VDSP) Intercomparison Study 2 - Part 2 ligand binding assays - impact of 25-hydroxyvitamin D₂ and 24R,25-dihydroxyvitamin D₃ on assay performance

Affiliations

Interlaboratory comparison of 25-hydroxyvitamin D assays: Vitamin D Standardization Program (VDSP) Intercomparison Study 2 - Part 2 ligand binding assays - impact of 25-hydroxyvitamin D₂ and 24R,25-dihydroxyvitamin D₃ on assay performance

Authors

Affiliations

Abstract

Conflict of interest statement

Figures

Similar articles

Cited by

References

MeSH terms

Substances

Grants and funding

LinkOut - more resources

Full Text Sources

Medical

Abstract

Conflict of interest statement

Figures

Similar articles

Cited by

References

MeSH terms

Substances

Related information

Grants and funding

LinkOut - more resources

Full Text Sources

Medical