A fast and simple method for simultaneous measurements of 25(OH)D, 24, 25(OH)2D and the Vitamin D Metabolite Ratio (VMR) in serum samples by LC-MS/MS
A fast and simple method for simultaneous measurements of 25(OH)D, 24, 25(OH)2D and the Vitamin D Metabolite Ratio (VMR) in serum samples by LC-MS/MS
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DOI：
10.1016/j.cca.2017.08.024
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发表时间：
2017-10-01
影响因子：
5
通讯作者：
Cavalier, Etienne
中科院分区：
文献类型：
--
作者：
Fabregat-Cabello, Neus;Farre-Segura, Jordi;Cavalier, Etienne
背景：为了满足临床常规实验室的需求，需要快速、简单且可靠地测量主要维生素 D 代谢物。为了克服这些挑战，我们开发并验证了一种 LC-MS/MS 方法，用于定量 25-羟基维生素 D-2 和 D-3、表观 25-羟基维生素 D-3 和 24,25-二羟基维生素 D-3。 方法：样品制备基于 100 μL 患者血清的沉淀和离心，然后注入 LC-MS/MS 系统。来自维生素 D 标准化计划的样品 (n = 80) 和患者样品 (n = 281) 已与参考 LC-MS/MS 方法进行了比较。使用 NIST 和 Labquality 质量控制材料进行分析验证。结果：平均测定内和测定间不精确度分别 < 6.0 和 6.4%，平均回收率在 95-104% 范围内。 24,25(OH)(2)D-3 的 LOQ 为 0.5 μg/L，25(OH)D-3 和 epi-25(OH)D-3 的 LOQ 为 1.1 μg/L，25(OH)D-2 的 LOQ 为 1.7 μg/L。拟议方法与参考方法之间存在 3% 的偏差，满足维生素 D 标准化计划的建议。结论：我们提出了一种快速、简单、可靠且经济高效的方法，完全适合常规检测，可测量血清样品中 24,25-二羟基维生素 D 与 25-羟基维生素 D 的比率，即维生素 D 代谢物比率 (VMR)。
Background: Rapid, easy and reliable measurement of the major vitamin D metabolites is required in order to fulfill the needs of a clinical routine laboratory. To overcome these challenges, we have developed and validated a LC-MS/MS method for the quantification of 25-hydroxyvitamin D-2 and D-3, epi-25-hydroxyvitamin D-3 and 24,25-dihydroxyvitamin D-3.Methods: Sample preparation was based on precipitation and centrifugation of 100 mu L of patient serum, followed by injection into the LC-MS/MS system. Samples from Vitamin D Standardization Program (n = 80) and patient samples (n = 281) have been compared with a reference LC-MS/MS method. For the analytical validation NIST and Labquality quality control materials were used.Results: Mean intra-assay and inter-assay imprecision were < 6.0 and 6.4% and mean recoveries were within 95-104%. LOQ's were 0.5 mu g/L for 24,25(OH)(2)D-3, 1.1 mu g/L for 25(OH)D-3 and epi-25(OH)D-3 and 1.7 mu g/L for 25(OH)D-2. A 3% bias obtained between the proposed and the reference method satisfies Vitamin D Standardization Program recommendations.Conclusions: We present a rapid, easy, reliable and cost-effective method completely adequate for routine testing, which permits the measurement of the ratio of 24,25-dihydroxyvitamin D to 25-hydroxyvitamin D, Vitamin D Metabolite Ratio (VMR), in serum samples.