CHROMATOGRAPHIC SEPARATION OF ESTRIOL, 16-KETOESTRADI OL-17β, 16-EPI ESTRI OL, ESTRADIOL-17α, ESTRADIOL-17β, ESTRONE AND 2-METHOXYESTRONE ON THE COLUMN OF PARTIALLY ESTERIFIED CARBOXYLIC ACID TYPE ION EXCHANGE RESIN

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Abstract
The adsorption chromatography of estrogens on the partially esterified carboxylic acid type ion exchange resin Amberlite IRC-50 was described. Estriol, 16-ketoestradiol-17[beta], 16-epiestriol, estradiol-17[alpha] estradiol-17[beta], estrone and 2-methoxyestrone could be separated by using 3 solvent systems. The mixture of estrogens was first fractionated by using a mixture of ethanol and water (32 by vol.) as the eluant and this eluant provided a good separation of estriol, 16-ketoestradiol-17[beta], 16-epiestriol, estradiol-17[alpha] plus estradiol-17[beta] and estrone plus 2-methoxyestrone and they were eluted in the order described. Then, the components that were eluted in the same fraction were separated by using a mixture of methanol, ethanol and water (3:15:8 by vol.) and of isopropanol and water (15:8 by vol.) as eluants. The former eluant separated estradiol-17[alpha] from estradiol-17[beta] at 15[degree] C and the latter eluant partially separated 2-methoxyestrone from estrone. The recovery of each estrogen was estimated and found to be 88 to 100%. The column could be used repeatedly since the condition of the column is unchanged after chromatography.