Analysis of vitamin D metabolites by liquid chromatography-tandem mass spectrometry

Measurement of vitamin D has increased significantly over the last years due to its involvement not only in calcium homeostasis, but also in the pathogenesis of various autoimmune- and infectious diseases. In the majority of studies, emphasis has been on 25-hydroxyvitamin D, which is regarded the best indicator of vitamin D status, whereas recently more interest has arisen in a combined measurement of multiple clinically relevant vitamin D metabolites. LC-MS/MS is a powerful technique allowing simultaneous determination of various vitamin D metabolites covering a broad dynamic range. Multi-panel LC-MS/MS assays are likely to enhance future investigations regarding the optimal combination of metabolites for the assessment of vitamin D sufficiency as well as help us in gaining better understanding of vitamin metabolism in health and disease. This article reviews recent advances in the quantification of the major vitamin D metabolites by LC-MS/MS and their application in clinical research.