Improved accuracy of an tandem liquid chromatography–mass spectrometry method measuring 24R,25-dihydroxyvitamin D3 and 25-hydroxyvitamin D metabolites in serum using unspiked controls and its application to determining cross-reactivity of a chemiluminescent microparticle immunoassay

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dc.contributor.authorDowling, Kirsten G.
dc.contributor.authorHull, George
dc.contributor.authorSundvall, Jouko
dc.contributor.authorLamberg-Allardt, Christel
dc.contributor.authorCashman, Kevin D.
dc.contributor.funderSeventh Framework Programmeen
dc.contributor.funderTerveyden ja hyvinvoinnin laitosen
dc.date.accessioned2021-08-25T08:32:02Z
dc.date.available2021-08-25T08:32:02Z
dc.date.issued2017-03-23
dc.date.updated2021-08-25T08:20:40Z
dc.description.abstractMeasurement of serum 25-hydroxyvitamin D [25(OH)D] is considered the best indicator of vitamin D status. Two minor vitamin D metabolites are common interferences encountered in 25(OH)D assays. The first is 3-epi-25-hydroxyvitamin D3 [3-epi-25(OH)D3], which if not chromatographically resolved from 25-hydroxyvitamin D3 [25(OH)D3], can overestimate 25(OH)D concentrations. The second is 24R,25-dihydroxyvitamin D3 [24R,25(OH)2D3], which can cross-react with the antibodies in 25(OH)D immunoassays. Our aim was to develop an LCâ MS/MS method capable of detecting both 3-epi-25(OH)D3 and 24R,25(OH)2D3 in serum without the use of a derivatization agent. We report an isotope dilution LCâ MS/MS method, with electrospray ionization in the positive mode, that can simultaneously detect 24R,25(OH)2D3, 25(OH)D3, 3-epi-25(OH)D3, and 25-hydroxyvitamin D2. The method employs a cost-effective liquidâ liquid extraction using only 150 μL of sera and a total run time of 10 min. Method performance was assessed by using quality controls made from pooled sera as an alternative to sera spiked with analytes. Biobanked samples, originally analyzed by chemiluminescent microparticle immunoassay (CMIA), were re-analyzed with this method to determine the contribution of 24R,25(OH)2D3 cross-reactivity to 25(OH)D measurement bias. The CMIA over-estimation of 25(OH)D measurements relative to LCâ MS/MS was found to depend on both 25(OH)D and 24R,25(OH)2D3 concentrations.en
dc.description.statusPeer revieweden
dc.description.versionAccepted Versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.citationDowling, K. G., Hull, G., Sundvall, J., Lamberg-Allardt, C. and Cashman, K. D. (2017) 'Improved accuracy of an tandem liquid chromatography–mass spectrometry method measuring 24R,25-dihydroxyvitamin D3 and 25-hydroxyvitamin D metabolites in serum using unspiked controls and its application to determining cross-reactivity of a chemiluminescent microparticle immunoassay', Journal of Chromatography A, 1497, pp. 102-109. doi: 10.1016/j.chroma.2017.03.058en
dc.identifier.doi10.1016/j.chroma.2017.03.058en
dc.identifier.endpage109en
dc.identifier.issn0021-9673
dc.identifier.journaltitleJournal of Chromatography Aen
dc.identifier.startpage102en
dc.identifier.urihttps://hdl.handle.net/10468/11780
dc.identifier.volume1497en
dc.language.isoenen
dc.publisherElsevier B.V.en
dc.relation.projectinfo:eu-repo/grantAgreement/EC/FP7::SP1::KBBE/613977/EU/Food-based solutions for Optimal vitamin D Nutrition and health through the life cycle/ODINen
dc.relation.urihttp://www.sciencedirect.com/science/article/pii/S0021967317304533
dc.rights© 2017, Elsevier B.V. All rights reserved. This manuscript version is made available under the CC BY-NC-ND 4.0 license.en
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/4.0/en
dc.subjectVitamin D metabolitesen
dc.subject25-hydroxyvitamin Den
dc.subjectQuality controlen
dc.subjectImmunoassayen
dc.subjectCross-reactivityen
dc.subjectLC–MS/MSen
dc.titleImproved accuracy of an tandem liquid chromatography–mass spectrometry method measuring 24R,25-dihydroxyvitamin D3 and 25-hydroxyvitamin D metabolites in serum using unspiked controls and its application to determining cross-reactivity of a chemiluminescent microparticle immunoassayen
dc.typeArticle (peer-reviewed)en
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