Mass spectrometry today is the only analytical technology which allows the specific determination of all known vitamin D metabolites. During the last few years the number of published methods rapidly increased and quantitative HPLC-MS/MS procedures are described for all the major metabolites including 25(OH)D3, 3-epi25(OH)D3, 24,25(OH)(2)D3, 25(OH)D2 and 1,25(OH)(2)D3. For the first time these new methods have made the systematic study of the clinical relevance of vitamin D metabolites possible. In parallel to the development of methods for new metabolites, significant progress was made in improving the performance of HPLC-MS/MS for quantifying 25(OH)D3 concentrations which has resulted in very short run times and thus enabling high throughput analysis for routine use and large sample sets.
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