Determination of Micro Amounts of Estrogens in Anabolic Vitamin Tablets by Quantitative Paper Chromatography

A quantitative paper chromatographic method has been developed which is used routinely as a stability assay for estrogenic hormones in anabolic vitamin tablets. The method involves ( a ) extraction of the estrogenic hormone into ether from an alcohol-water suspension of the ground tablets, ( b ) clarification of the extract by chromatography on an alumina column, ( c ) paper chromatographic separation of the estrogenic hormone present in the column eluate from interfering degradation products, ( d ) location of the estrogenic hormone on the chromatogram by the guide strip technique employing a chromogenic agent, ( e ) elution of the estrogenic hormone from the chromatogram, and ( f ) quantitative spectrophotofluorometric analysis of the eluate. Recoveries in excess of 95 per cent are obtained by chromatographing simultaneously replicates of standard and sample solutions on the same chromatogram. A detailed account of the procedure is presented using the assay of ethinyl estradiol in concentrations of 0.0014 per cent in anabolic vitamin tablets as an example.