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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
biochemistry, genetics and molecular biology
Quantitative determination of vitamin D metabolites in plasma using UHPLC-MS/MS
Analytical and Bioanalytical Chemistry, Volume 398, No. 2, Year 2010
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Description
Vitamin D is an important determinant of bone health at all ages. The plasma concentrations of 25-hydroxy vitamin D (25-OH D) and other metabolites are used as biomarkers for vitamin sufficiency and function. To allow for the simultaneous determination of five vitamin D metabolites, 25-OH D3, 25-OH D2, 24,25-(OH)2 D3, 1,25-(OH)2 D3, and 1,25-(OH)2 D2, in low volumes of human plasma, an assay using ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was established. Plasma samples were spiked with isotope-labeled internal standards and pretreated using protein precipitation, solid-phase extraction (SPE) and a Diels-Alder derivatization step with 4-phenyl-1,2,4-triazoline-3,5-dione. The SPE recovery rates ranged from 55% to 85%, depending on the vitamin D metabolite; the total sample run time was <5 min. Mass spectrometry was conducted using positive ion electrospray ionization in the multiple reaction monitoring mode on a quadrupole-quadrupole-linear ion trap instrument after pre-column addition of methylamine to increase the ionization efficiency. The intra- and inter-day relative standard deviations were 1.6-4.1% and 3.7-6.8%, respectively. The limit of quantitation for these compounds was determined to be between 10 and 20 pg/mL. The 25-OH D results were compared with values obtained for reference materials (DEQAS). In addition, plasma samples were analyzed with two additional Diasorin antibody assays. All comparisons with conventional methods showed excellent correlations (r 2∈=∈0.9738) for DEQAS samples, demonstrating the high degree of comparability of the new UHPLC-MS/MS technique to existing methods. © 2010 The Author(s).
Authors & Co-Authors
Ding, Shujing
United Kingdom, London
Medical Research Council
Schoenmakers, Inez
United Kingdom, London
Medical Research Council
Jones, Kerry S.
Gambia, Banjul
Medical Research Council Laboratories Gambia
Koulman, Albert
United Kingdom, London
Medical Research Council
Prentice, Ann M.
United Kingdom, London
Medical Research Council
Volmer, Dietrich A.
United Kingdom, London
Medical Research Council
Germany, Saarbrucken
Universität Des Saarlandes
Statistics
Citations: 157
Authors: 6
Affiliations: 3
Identifiers
Doi:
10.1007/s00216-010-3993-0
ISSN:
16182642
Study Approach
Quantitative