A QUANTITATIVE STUDY OF THE ADSORPTION OF ESTRONE, ESTRIOL, AND cr-ESTRADIOL ON A CHROMATOGRAPHIC

In the course of a study of various procedures for the concentration and purification of urinary estrogens, the feasibility of the chromatographic technique seemed to warrant investigation. Chromatography has been employed to advantage in isolation procedures (l-3) for certain estrogens but no attempt has been made to utilize it in any quantitative procedure for estrone, estriol, and cY-estradiol, the three estrogens believed to occur in human female urine (4,5). Furthermore, it was suspected that sufficient differences in adsorptive characteristics might exist among the three estrogens to effect a reasonably accurate separation from a mixture. Current procedures for the quantitative separation of mixtures of the three naturally occurring estrogens depend upon differences in distribution between immiscible solvents (6-8) or upon differences in chemical reactivity of the C-17 substituent group (9, 10). We have investigated the adsorption characteristics of crystalline estrone, estriol, and a-e&radio1 on a column of activated alumina and have developed a chromatographic procedure for the quantitative separation of the strongly phenolic estrogen (estriol) from the weakly phenolic estrogens (estrone and cu-estradiol). This investigation and procedure form the subject of this report. The application of the chromatographic technique to suitable urinary estrogenic residues will be the subject of a forthcoming publication.