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Journal of Lipid Research, Vol. 15, 517-524, September 1974
Copyright © 1974 by Lipid Research, Inc.

Liquid-gel partition chromatography of vitamin A compounds; formation of retinoic acid from retinyl acetate in vivo

Y. L. Ito , M. Zile , H. Ahrens , and H. F. DeLuca

Department of Biochemistry, College of Agricultural and Life Sciences, University of Wisconsin-Madison, Madison, Wisconsin 53706

A clear separation of retinol, retinal, and retinoic acid has been achieved by liquid-gel partition chromatography on Sephadex LH-20 with solvent mixtures of chloroform, Skellysolve B, and methanol. A mixture of retinyl esters, retinol, retinal, and retinoic acid has been resolved on hydroxyalkoxypropyl Sephadex using Skellysolve B and acetone. There is no decomposition of any of the vitamin A compounds during chromatography, and recovery is complete. The combination of mildness and potential for resolution makes liquid-gel partition chromatography a superior tool for the separation of vitamin A compounds. This method has been applied to the study of vitamin A metabolism at physiological levels in the vitamin A-deficient rat. Retinyl palmitate, an ester of retinoic acid, retinal, retinol, retinoic acid, and a polar metabolite have been demonstrated in various tissues of the rat 12 hr after a dose of 2 µg of [11-¹⁴C]retinyl acetate.

Supplementary key words retinol • retinal • retinyl esters • Sephadex LH-20 • hydroxyalkoxypropyl Sephadex • rat

Submitted on March 13, 1974
Accepted on June 6, 1974

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