Investigation of C-3 epimerization mechanism of 24,25-dihydroxyvitamin D3 in rat using liquid chromatography/mass spectrometry

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Studies on C-3 epimerization of 24,25-dihydroxyvitamin D3 [24,25(OH)2D3] were performed. 3-Epi-24,25(OH)2D3 was found to circulate in the blood of rats administered 24,25(OH)2D3, which was confirmed in comparison with a standard sample based on its chromatographic behavior during inclusion high-performance liquid chromatography using γ-cyclodextrin as a mobile phase additive and data obtained from liquid chromatography (LC)/mass spectrometry (MS) and gas chromatography/MS. The C-3 epimerization mechanism was investigated in in vitro experiments using [3α-2H]-24,25(OH)2D3. The disappearance of deuterium was monitored by LC/MS. As a result, it was clarified that the 3-epimer was formed via the 3-oxo-form as an intermediate. This epimerization was also found to be controlled by the enzyme existing in the liver cytosol fraction and to require NAD and NADPH as coenzymes.
Japan Society for Analytical Chemistry = 日本分析化学会の論文
2000-05-10