Abstract
An analytical method was developed for the identification of primary vitamin D3 metabolites in human urine using liquid chromatography tandem mass spectrometry in positive mode. Urine samples were purified using C18 solid-phase extraction cartridges and analytical separations were performed by reversed phase liquid chromatography in gradient mode using ammonium acetate (0.01 mol L−1) and acetonitrile as the mobile phases. Identification and structural elucidation of the metabolites were carried out by comparison with mass spectral fragmentation behavior of vitamin D3 and retention characteristics. Three primary urinary vitamin D3 metabolites were identified as 25-hydroxyvitamin D3, 1α,25-dihydroxyvitamin D3 and vitamin D3 sulphate, respectively.
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