Isolation and identification of 25-hydroxycholecalciferol from human plasma.

Two polar metabolites of vitamin D3have been isolated from the plasma of a hypoparathyroid patient that had been treated with 1.5 mg of vitamin D3per day. One of the metabolites (peak IV) has been conclusively identified as 25-hydroxycholecalciferol (25-HCC) by means of mass spectrometry, ultraviolet absorption spectrophotometry, and gas-liquid chromatography. Furthermore, this metabolite has the same potency as 25-HCC in the cure of rickets in rats. The second metabolite (peak V) was identified as 21,25-dihydroxycholecalciferol (21,25-DHCC) on the basis of its ultraviolet absorption spectrum and cochromatography with authentic 21,25-DHCC in three different chromatographic systems.

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