Abstract

The separation of side chain dihydroxylated vitamin D3 metabolites by inclusion high performance liquid chromatography using cyclodextrin (CD) as the mobile phase additive was examined. 24,25-Dihydroxyvitamin D3 and its 3-epimer were clearly separated by the addition of γ-CD to the mobile phase using acetonitrile as the organic modifier. The separation of the 25-epimers of 25,26-dihydroxyvitamin D3 has been done, to a certain extent, with the mobile phase using methanol and dimethylated β-CD as the organic modifier and the additive, respectively. These separation methods were applied to the detection of the metabolites in the plasma of the rat administered 25-hydroxyvitamin D3. These metabolites were also identified by liquid chromatography/tandem mass spectrometry after derivatization.

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