Isolation, identification, and biological activity of 25-hydroxy-24-oxovitamin D3: a new metabolite of vitamin D3 generated by in vitro incubations with kidney homogenates

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Abstract
A metabolite of 25-hydroxyvitamin D3 was isolated in pure form from incubation mixtures containing kidney homogenates of chicks given large doses of vitamin D3. The isolation involved methanol-chloroform extraction and 6 steps of column chromatography. The metabolite was identified as 25-hydroxy-24-ovovitamin D3 by means of UV absorption spectrometry, mass spectrometry, IR spectrometry, NMR spectrometry and specific chemical reactions. Use of a sensitive in situ technique revealed that 25-hydroxy-24-oxovitamin D3 enhances intestinal Ca transport in rats approximately as effectively as 24,25-dihydroxyvitamin D3 does. In contrast, 25-hydroxy-24-oxovitamin D3 appeared to be less active than 24,25-dihydroxyvitamin D3 in chicks 24 h after IV injection.