title: Triple Quadrupole Versus High Resolution Quadrupole-Time-of-Flight Mass Spectrometry for Quantitative LC-MS/MS Analysis of 25-Hydroxyvitamin D in Human Serum.
abstract: We describe a systematic comparison of high and low resolution LC-MS/MS assays for quantification of 25-hydroxyvitamin D3 in human serum. Identical sample preparation, chromatography separations, electrospray ionization sources, precursor ion selection, and ion activation were used; the two assays differed only in the implemented final mass analyzer stage; viz. high resolution quadrupole-quadrupole-time-of-flight (QqTOF) versus low resolution triple quadrupole instruments. The results were assessed against measured concentration levels from a routine clinical chemiluminescence immunoassay. Isobaric interferences prevented the simple use of TOF-MS spectra for extraction of accurate masses and necessitated the application of collision-induced dissociation on the QqTOF platform. The two mass spectrometry assays provided very similar analytical figures of merit, reflecting the lack of relevant isobaric interferences in the MS/MS domain, and were successfully applied to determine the levels of 25-hydroxyvitamin D for patients with chronic liver disease. Graphical Abstract ᅟ.
authors.affiliation: Institute of Bioanalytical Chemistry, Saarland University, Saarbrücken, Germany.
authors.forename: Timon
authors.initials: T
authors.lastname: Geib
authors.affiliation: Department of Chemistry, Université du Québec à Montréal, Montréal, Québec, Canada.
authors.forename: Lekha
authors.initials: L
authors.lastname: Sleno
authors.affiliation: Department of Medicine II, Saarland University Medical Center, Homburg, Germany.
authors.forename: Rabea A
authors.initials: RA
authors.lastname: Hall
authors.affiliation: Department of Medicine II, Saarland University Medical Center, Homburg, Germany.
authors.forename: Caroline S
authors.initials: CS
authors.lastname: Stokes
authors.affiliation: Institute of Bioanalytical Chemistry, Saarland University, Saarbrücken, Germany. dietrich.volmer@mx.uni-saarland.de.
authors.forename: Dietrich A
authors.initials: DA
authors.lastname: Volmer
mesh_terms: D002853:Chromatography, Liquid; D006801:Humans; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry; D014807:Vitamin D / Q000031:analogs & derivatives* / Q000097:blood
chemical_list: D014807:Vitamin D; C104450:25-hydroxyvitamin D
keywords: Collision-induced dissociation; Electrospray ionization; High resolution mass spectrometry; Isobaric interferences; Vitamin D