Institute of Metabolism and Systems Research, the University of Birmingham, Birmingham B15 2TT, UK.
Institute of Metabolism and Systems Research, the University of Birmingham, Birmingham B15 2TT, UK.
J Chromatogr B Analyt Technol Biomed Life Sci. 2018 Jun 15;1087-1088:43-48. doi: 10.1016/j.jchromb.2018.04.025. Epub 2018 Apr 18.
In recent years, increased interest in the human health benefits of vitamin D has led to demand for improved analysis of patient vitamin D 'status'. Studies to date have focused primarily on a single vitamin D metabolite, 25-hydroxyvitamin D, despite the existence of a broad range of vitamin D metabolites, referred to as the vitamin D metabolome. This study reports on the development of a rapid UPSFC-MS/MS method for the analysis of nine vitamin D metabolites in human serum. Optimum separation was obtained with a Lux-Cellulose chiral column. We observed an orthogonal elution order when compared with ultra-high performance liquid chromatography (UHPLC). The order of elution was reversed based on hydroxyl- group number, however elution order did not differ between isomeric changes in hydroxyl- group position or epimers. Although UPSFC yielded superior resolution and selectivity over previously developed UHPLC-MS/MS methods, improvements in sensitivity could not be achieved owing to the lower injection volume required for UPSFC relative to UHPLC. Method validation was performed on the developed UPSFC-MS/MS method and found to be within acceptable limits. Applying the method to the analysis of human serum samples showed a significant correlation with serum concentrations of metabolites measured by UHPLC-MS/MS (25OHD3 r = 0.997, P=<0.001, and 3-epi-25OHD3 r = 0.996, P ≤0.001). These data indicate that UPSFC provides an efficient analytical platform for rapid analysis of multiple vitamin D metabolites from serum.
近年来，人们对维生素 D 对人体健康益处的兴趣日益增加，这导致人们对改善患者维生素 D“状况”的分析提出了更高的要求。迄今为止，研究主要集中在单一的维生素 D 代谢物 25-羟维生素 D 上，尽管存在广泛的维生素 D 代谢物，被称为维生素 D 代谢组。本研究报告了一种快速 UPSFC-MS/MS 方法的开发，用于分析人血清中的九种维生素 D 代谢物。最优分离是在 Lux-Cellulose 手性柱上实现的。与超高效液相色谱（UHPLC）相比，我们观察到正交洗脱顺序。然而，在羟基位置的异构变化或差向异构体之间，洗脱顺序没有差异。尽管 UPSFC 相对于先前开发的 UHPLC-MS/MS 方法具有更高的分辨率和选择性，但由于 UPSFC 相对于 UHPLC 需要的进样体积较小，因此无法实现灵敏度的提高。对开发的 UPSFC-MS/MS 方法进行了方法验证，发现结果在可接受范围内。将该方法应用于人血清样品的分析表明，与 UHPLC-MS/MS 测量的代谢物血清浓度具有显著相关性（25OHD3 r=0.997，P<0.001，3-epi-25OHD3 r=0.996，P≤0.001）。这些数据表明，UPSFC 为快速分析血清中的多种维生素 D 代谢物提供了一个有效的分析平台。
