Interlaboratory comparison of 25-hydroxyvitamin D assays: Vitamin D Standardization Program (VDSP) Intercomparison Study 2 — Part 1 liquid chromatography – tandem mass spectrometry (LC-MS/MS) assays — impact of 3-epi-25-hydroxyvitamin 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, Andrew N. Hoofnagle, Fiona Ivison, Ralf Fischer, Jody M.W. van den Ouweland, Chung S. Ho, Emmett W.K. Law, Jean Nicolas Simard, Renaud Gonthier, Brett Holmquist, Sarah Meadows, Lorna CoxKimberly Robyak, Michael H. Creer, Robert Fitzgerald, Michael W. Clarke, Norma Breen, Pierre Lukas, Étienne Cavalier, Christopher T. Sempos

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Abstract

An interlaboratory comparison study was conducted by the Vitamin D Standardization Program (VDSP) to assess the performance of liquid chromatography – tandem mass spectrometry (LC-MS/MS) assays used for the determination of serum total 25-hydroxyvitamin D (25(OH)D), which is the sum of 25-hydroxyvitamin D2 (25(OH)D2) and 25-hydroxyvitamin D3 (25(OH)D3). A set of 50 single-donor samples was assigned target values for concentrations of 25(OH)D2, 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 1 includes results from 14 laboratories using 14 custom LC-MS/MS 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 53% of the LC-MS/MS assays met the VDSP criterion of mean % bias ≤ |±5%|. For the LC-MS/MS assays not meeting the ≤ |±5%| criterion, four assays had mean % bias of between 12 and 21%. Based on multivariable regression analysis using the concentrations of the four individual vitamin D metabolites in the 50 single-donor samples, the performance of several LC-MS/MS assays was found to be influenced by the presence of 3-epi-25(OH)D3. The results of this interlaboratory study represent the most comprehensive comparison of LC-MS/MS assay performance for serum total 25(OH)D and document the significant impact of the lack of separation of 3-epi-25(OH)D3 and 25(OH)D3 on assay performance, particularly with regard to mean % bias. Graphical abstract: [Figure not available: see fulltext.].

Original languageEnglish (US)
Pages (from-to)333-349
Number of pages17
JournalAnalytical and Bioanalytical Chemistry
Volume414
Issue number1
DOIs
StatePublished - Jan 2022

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry

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