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; Johanna E Camara; Carolyn Q Burdette; Grace Hahm; Federica Nalin; Adam J Kuszak; Joyce Merkel; Ramón A Durazo-Arvizu; Emma L Williams; Christian Popp; Christian Beckert; Jan Schultess; Glen Van Slooten; Carole Tourneur; Camille Pease; Ravi Kaul; Alfredo Villarreal; Marcelo Cidade Batista; Heather Pham; Alex Bennett; Eugene Jansen; Dilshad Ahmed Khan; Mark Kilbane; Patrick J Twomey; James Freeman; Neil Parker; Sohail Mushtaq; Christine Simpson; Pierre Lukas; Étienne Cavalier; Christopher T Sempos

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

Anal Bioanal Chem. 2022 Jan;414(1):351-366. doi: 10.1007/s00216-021-03577-0. Epub 2021 Aug 25.

Authors

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.

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].

MeSH terms

25-Hydroxyvitamin D 2
Chromatography, Liquid
Ligands
Reference Standards
Tandem Mass Spectrometry*
Vitamin D* / analogs & derivatives

Substances

Ligands
Vitamin D
25-Hydroxyvitamin D 2
25-hydroxyvitamin D

Abstract

MeSH terms

Substances

Grants and funding