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2023-05-23Zeitschriftenartikel DOI: 10.18452/28524
Comparing derivatization reagents for quantitative LC–MS/MS analysis of a variety of vitamin D metabolites
dc.contributor.authorAlexandridou, Anastasia
dc.contributor.authorSchorr, Pascal
dc.contributor.authorVolmer, Dietrich
dc.date.accessioned2024-04-17T15:11:43Z
dc.date.available2024-04-17T15:11:43Z
dc.date.issued2023-05-23none
dc.identifier.urihttp://edoc.hu-berlin.de/18452/29153
dc.description.abstractThe present study systematically compares the sensitivity and selectivity of the analysis of multiple vitamin D metabolites after chemical derivatization using different reagents for liquid chromatography-tandem mass spectrometry (LC–MS/MS). Generally, chemical derivatization is applied to vitamin D metabolites to increase the ionization efficiency, which is particularly important for very low abundant metabolites. Derivatization can also improve the selectivity of the LC separation. A wide variety of derivatization reagents has been reported in recent years, but information on their relative performance and applicability to different vitamin D metabolites is, unfortunately, not available in the literature. To fill this gap, we investigated vitamin D3, 3β-25-hydroxyvitamin D3 (3β-25(OH)D3), 3α-25-hydroxyvitamin D3 (3α-25(OH)D3), 1,25-dihydroxyvitamin D3 (1,25(OH)2D3), and 24,25-dihydroxyvitamin D3 (24,25(OH)2D3) and compared response factors and selectivity after derivatizing with several important reagents, including four dienophile reagents (4-phenyl-1,2,4-triazoline-3,5-dione (PTAD), 4-[2-(6,7-dimethoxy-4-methyl-3-oxo-3,4-dihydroquinoxalinyl)ethyl]-1,2,4-triazoline-3,5-dione (DMEQ-TAD), Amplifex, 2-nitrosopyridine (PyrNO)) as well as two reagents targeting hydroxyl groups: isonicotinoyl chloride (INC) and 2-fluoro-1-methylpyridinium-p-toluenesulfonate (FMP-TS). In addition, a combination of dienophiles and hydroxyl group reagents was examined. For LC separations, reversed-phase C-18 and mixed-mode pentafluorophenyl HPLC columns using different compositions of the mobile phase were compared. With respect to detection sensitivity, the optimum derivatization reagent for the profiling of multiple metabolites was Amplifex. Nevertheless, FMP-TS, INC, PTAD, or PTAD combined with an acetylation reaction showed very good performance for selected metabolites. These reagent combinations provided signal enhancements on the order of 3- to 295-fold depending on the compound. Chromatographic separation of the dihydroxylated vitamin D3 species was readily achieved using any of the derivatization reactions, while for 25(OH)D3 epimers, only PyrNO, FMP, INC, and PTAD combined with acetylation enabled complete separation. In conclusion, we believe this study can serve as a useful reference for vitamin D laboratories, to help analytical and clinical scientists decide which derivatization reagent to choose for their application.eng
dc.language.isoengnone
dc.publisherHumboldt-Universität zu Berlin
dc.rights(CC BY 4.0) Attribution 4.0 Internationalger
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.subjectVitamin D3 metaboliteseng
dc.subject25-Hydroxyvitamin D3eng
dc.subjectLC–MS/MSeng
dc.subjectElectrosprayeng
dc.subjectChemical derivatizationeng
dc.subjectEpimerseng
dc.subject.ddc540 Chemie und zugeordnete Wissenschaftennone
dc.titleComparing derivatization reagents for quantitative LC–MS/MS analysis of a variety of vitamin D metabolitesnone
dc.typearticle
dc.identifier.urnurn:nbn:de:kobv:11-110-18452/29153-7
dc.identifier.doihttp://dx.doi.org/10.18452/28524
dc.type.versionpublishedVersionnone
local.edoc.pages13none
local.edoc.type-nameZeitschriftenartikel
local.edoc.container-typeperiodical
local.edoc.container-type-nameZeitschrift
dc.description.versionPeer Reviewednone
dc.identifier.eissn1618-2650
dcterms.bibliographicCitation.doi10.1007/s00216-023-04753-0
dcterms.bibliographicCitation.journaltitleAnalytical and bioanalytical chemistrynone
dcterms.bibliographicCitation.volume415none
dcterms.bibliographicCitation.issue19none
dcterms.bibliographicCitation.originalpublishernameSpringernone
dcterms.bibliographicCitation.originalpublisherplaceHeidelbergnone
dcterms.bibliographicCitation.pagestart4689none
dcterms.bibliographicCitation.pageend4701none
bua.departmentMathematisch-Naturwissenschaftliche Fakultätnone

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