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

被引:30
作者
Dowling, Kirsten G. [1 ]
Hull, George [1 ]
Sundvall, Jouko [2 ]
Lamberg-Allardt, Christel [3 ]
Cashman, Kevin D. [1 ,4 ]
机构
[1] Univ Coll Cork, Sch Food & Nutr Sci, Cork Ctr Vitamin & Nutr Res D, Cork, Ireland
[2] Natl Inst Hlth & Welf THL, Genom & Biomarkers Unit, Dept Hlth, Helsinki, Finland
[3] Univ Helsinki, Dept Food & Environm Sci, Helsinki, Finland
[4] Univ Coll Cork, Dept Med, Cork, Ireland
关键词
Vitamin D metabolites; 25-hydroxyvitamin D; LC-MS/MS; Quality control; Immunoassay; Cross; -reactivity; VITAMIN-D METABOLITES; STANDARDIZATION PROGRAM PROTOCOLS; LC-MS/MS METHOD; D DEFICIENCY; 24,25-DIHYDROXYVITAMIN D; QUANTIFICATION; DERIVATIZATION; PREVENTION; NUTRITION; HEALTH;
D O I
10.1016/j.chroma.2017.03.058
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:102 / 109
页数:8
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