J. Lipid Res. Neurobiology of Lipids (ISSN1683-5506)
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Journal of Lipid Research, Vol. 12, 460-465, July 1971
Copyright © 1971 by Lipid Research, Inc.

A new chromatographic system for vitamin D3 and its metabolites: resolution of a new vitamin D3 metabolite

M. F. Holick and H. F. DeLuca

Department of Biochemistry, University of Wisconsin, Madison, Wisconsin 53706

A simple yet powerful new chromatographic procedure for vitamin D3 and its metabolites is described. Liquid-gel partition chromatography on Sephadex LH-20 using a solvent of various percentages of CHCl3 in Skellysolve B (petroleum ether, bp 67-69°C) permits excellent resolution of vitamin D3, 25-hydroxyvitamin D3, and their more polar metabolites. Of special importance is the resolution of the metabolites of vitamin D3 more polar than 25-hydroxycholecalciferol. Because of this resolution, a new metabolite of vitamin D3 has been demonstrated in the plasma of rats and in the intestines of chicks given 100 IU of vitamin D3-1,2-3H.

Supplementary key words Sephadex LH-20 • cholecalciferol • intestine • plasma • chick • rat • 25-hydroxycholecalciferol

Submitted on December 11, 1970
Accepted on March 3, 1971


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