Abstract
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.
Original language | English |
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Pages (from-to) | 43-48 |
Journal | Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences |
Volume | 1087-1088 |
Early online date | 18 Apr 2018 |
DOIs | |
Publication status | E-pub ahead of print - 18 Apr 2018 |
Keywords
- vitamin D
- UPSFC-MS/MS
- supercritical fluid chromatography-tandem mass spectrometry