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
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
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DOI：
10.1016/j.chroma.2017.03.058
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发表时间：
2017-05-12
影响因子：
4.1
通讯作者：
Cashman, Kevin D.
中科院分区：
文献类型：
--
作者：
Dowling, Kirsten G.;Hull, George;Cashman, Kevin D.
血清25-羟基维生素D[25(OH)D]的测定被认为是维生素D状态的最佳指标。在25(OH)D测定中，两种次要的维生素D代谢物是常见的干扰。第一种是3-epi-25-羟基维生素D-3[3-epi-25(OH)D-3]，如果不能从25-羟基维生素D-3[25(OH)D-3]中分离出来，可能会高估25(OH)D的浓度。第二种是24R，25-二羟基维生素D-3[24R，25(OH)(2)D-3]，它能与25(OH)D免疫分析中的抗体发生交叉反应。目的是建立一种无需衍生化试剂即可同时检测血清中3-epi-25(OH)D-3和24R，25(OH)(2)D-3的LC-MS/MS方法。报道了一种电喷雾正电离的同位素稀释LC-MS/MS方法，可以同时检测24R，25(OH)(2)D-3，25(OH)D-3，3-epi-25(OH)D-3和25-羟基维生素D-2。该方法采用高性价比的液-液萃取法，只需150亩L血清，总运行时间为10min。方法通过使用混合血清作为添加分析物的血清的替代品来评估性能。用这种方法重新分析了原来用化学发光微粒子免疫分析(CMIA)分析的生物库样品，以确定24R，25(OH)(2)D-3交叉反应对25(OH)D测量偏差的贡献。与LC-MS/MS相比，CMIA对25(OH)D测量值的高估与25(OH)D和24R，25(OH)(2)D-3的浓度有关。(C)2017爱思唯尔B.V.保留所有权利。
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.