Journal
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Volume 414, Issue 1, Pages 333-349Publisher
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-021-03576-1
Keywords
Total serum 25-hydroxyvitamin D (25(OH)D); 25-Hydroxyvitamin D-3 (25(OH)D-3); 25-Hydroxyvitamin D-2 (25(OH)D-2); 3-epi-25-Hydroxyvitamin D-3 (3-epi-25(OH)D-3); 24R,25-Dihydroxyvitamin D-3 (24R,25(OH)(3)D-3); Liquid chromatography - tandem mass spectrometry (LC-MS/MS)
Funding
- Office of Dietary Supplements at the National Institutes of Health (NIH-ODS)
- Western Australian State Government
- Australian Federal Government, through Bioplatforms Australia
- National Collaborative Research Infrastructure Strategy (NCRIS)
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The Vitamin D Standardization Program (VDSP) conducted an interlaboratory comparison study to evaluate the performance of liquid chromatography - tandem mass spectrometry (LC-MS/MS) assays for determining serum total 25-hydroxyvitamin D. Results showed that only 53% of the assays met the VDSP criterion, with four assays having significant performance issues due to the lack of separation between 3-epi-25(OH)D-3 and 25(OH)D-3.
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 D-2 (25(OH)D-2) and 25-hydroxyvitamin D-3 (25(OH)D-3). A set of 50 single-donor samples was assigned target values for concentrations of 25(OH)D-2, 25(OH)D-3, 3-epi-25-hydroxyvitamin D-3 (3-epi-25(OH)D-3), and 24R,25-dihydroxyvitamin D-3 (24R,25(OH)(2)D-3) 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)D-3. 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)D-3 and 25(OH)D-3 on assay performance, particularly with regard to mean % bias.
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