The colorimetric Determination of Estrone, Equilin and Equilenin in Mixtures

The Analysis of Conjugated Estrogen Preparations

“…2 Estrogens were purified and separated by the use of Girard's Reagent T, as directed by Carol and Rotondaro (11). Ketosteroids were analyzed according t o the differential procedure of Banes (12) and total diols were estimated in terms of estrone equivalent by the Kober procedure described in the preceding footnote. "Total estrogens" were calculated as the sum of estrone plus equilin plus equilenin plus total diols.…”

“…aliquots were evaporated and analyzed colorimetrically for "Kober chromogens" as estrone, and a 100-ml. aliquot was evaporated t o a small volume for the differential procedure (11,12). Procedure II.…”

The decomposition of equilin in oil solution**Parts of this paper have been presented by Daniel Banes in a dissertation offered in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the Department of Chemistry at Georgetown University.

“…The sesame oil solutions of estrogenic ketosteroids, whose analyses have been reported earlier (1) and are tabulated in Table 1 ("Recovered after 1 week"), were again extracted after several months and assayed according to the procedures previously described (1,2). Comparison of these analytical data (Table I, "Recovered after three months") with those first obtained showed a pronounced loss of equilixi.…”

“…Ketosteroids in 10-ml. portions of solution were extracted by the procedure of Carol with BQC reagent (1). The course of the decomposition of equilin in oil solutions exposed to light is shown graphically in Fig.…”

“…The decomposition is accelerated by light. N AN earlier report from this laboratory (1) I describing a colorimetric method for the analysis of estrogenic ketosteroid mixtures, i t was noted that recoveries of equilin were somewhat low when oil solutions of known concentration were assayed. It was suggested there that the losses could be ascribed t o the susceptibility of the ketosteroid t o chemical change.…”