Jongen M J, van der Vijgh W J, Willems H J, Netelenbos J C, Lips P
Clin Chem. 1981 Oct;27(10):1757-60.
We describe a simultaneous assay for the principal vitamin D metabolites: 25-hydroxyvitamin D, 24-25-dihydroxyvitamin D, and 1,25-dihydroxyvitamin D. Special attention has been paid to simplification of the extensive extraction and purification procedures used in previously described simultaneous assays. All three metabolites were isolated with a single extraction step, followed by only one gradient liquid-chromatographic procedure. For final quantitation we used competitive protein binding assays, involving readily available binding proteins and commercially purchased tritiated vitamin D metabolites. Concentrations in the plasma of healthy subjects (mean age, 27 years), sampled during December were 51 (SD 17) nmol/L, 4.1 (SD 1.3) nmol/L, and 124 (SD 26) pmol/L for 25-hydroxyvitamin D, 24,25-dihydroxyvitamin D and 1,25-dihydroxyvitamin D, respectively. Intra- and interassay CVs for the three metabolites were 4.4 and 3.9%, 6.7 and 8.0%, and 7.0 and 4.8%, respectively.
我们描述了一种针对主要维生素D代谢物的同步检测方法：25-羟基维生素D、24,25-二羟基维生素D和1,25-二羟基维生素D。特别关注了对先前描述的同步检测中广泛使用的提取和纯化程序的简化。所有三种代谢物通过单一提取步骤分离，随后仅进行一次梯度液相色谱程序。为了进行最终定量，我们使用了竞争性蛋白结合分析，涉及易于获得的结合蛋白和商业购买的氚标记维生素D代谢物。12月期间采集的健康受试者（平均年龄27岁）血浆中，25-羟基维生素D、24,25-二羟基维生素D和1,25-二羟基维生素D的浓度分别为51（标准差17）nmol/L、4.1（标准差1.3）nmol/L和124（标准差26）pmol/L。三种代谢物的批内和批间变异系数分别为4.4%和3.9%、6.7%和8.0%、7.0%和4.8%。
