The isolation and estimation of the steroid oestrogens in placental tissue

Publisher Website
Google Scholar
Abstract
A method was developed for isolating and estimating estrogens in placental tissue. It involves a preliminary chemical separation followed by paper chromatography, elution and fluorimetric estimation. The use of Folin and Ciocalteu''s reagent in a sensi-tive method for detection of the estrogens on paper chromatograms is described. The use of azo-dye coupling followed by paper chromatography was adapted to the quantitative assay of estrone and estradiol, and a new method of assay for estriol, estrone and estradiol using Folin and Ciocalteu''s reagent is described. The uncorrected avg. quantities of "free" estrogen/kg in 7 full-term placentae were; estriol, 190 [mu]g, estrone, 80 ug and estradiol, 25 [mu]g. Thevalues for "conjugated" estrogen were, respectively: 44, 21 and 8 [mu]g, and for "protein-bound" estrogen; 21, 14 and 7[mu]g. Large differences were found in the estrogen content of individual placentae. Recovery experiments show that the normal methods of extraction and purification, when applied to the "free" estrogens in full-term placental tissue give losses of between 70 and 90%. The corrected average quantities of "free" estrogen/ kg in 7 full-term placentae were estimated as: estriol, 680 [mu]g, estrone, 580 [mu]g and estradiol, 170 [mu]g/kg. While these corrections are accurate for the 7 placentae examined, they may be only roughly applicable to other samples of this tissue.