期刊
JOURNAL OF CHROMATOGRAPHY A
卷 1497, 期 -, 页码 102-109出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2017.03.058
关键词
Vitamin D metabolites; 25-hydroxyvitamin D; LC-MS/MS; Quality control; Immunoassay; Cross -reactivity
资金
- European Union's Seventh Framework Programme (FP7) [613977]
Measurement 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 D-3 [3-epi-25(OH)D-3], which if not chromatographically resolved from 25-hydroxyvitamin D-3 [25(OH)D-3], can overestimate 25(OH)D concentrations. The second is 24R,25-dihydroxyvitamin D-3 [24R,25(OH)(2)D-3], which can cross-react with the antibodies in 25(OH)D immunoassays. Our aim was to develop an LC-MS/MS method capable of detecting both 3-epi-25(OH)D-3 and 24R,25(OH)(2)D-3 in serum without the use of a derivatization agent. We report an isotope dilution LC-MS/MS method, with electrospray ionization in the positive mode, that can simultaneously detect 24R,25(OH)(2)D-3, 25(OH)D-3, 3-epi-25(OH)D-3, and 25-hydroxyvitamin D-2. The method employs a cost-effective liquid-liquid extraction using only 150 mu L 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)(2)D-3 cross reactivity to 25(OH)D measurement bias. The CMIA over-estimation of 25(OH)D measurements relative to LC-MS/MS was found to depend on both 25(OH)D and 24R,25(OH)(2)D-3 concentrations. (C) 2017 Elsevier B.V. All rights reserved.
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