Abstract
A fluorometric method to determine total urinary estrogens is presented. This procedure is based upon acid hydrolysis, solvent partition, reduction of estrone to estradiol, methylation and chromatography. The estrogens are measured by fluorometry of the Kober chromogens following chloroform extraction according to Ittrich. As estradiol- and estriol-3-methyl ether have a similar relative fluorescence intensity, estrone, estradiol and estriol are quantitated proportionally in a single fluorometric reading. All results are corrected for procedural loss by adding a 2:1:3 mixture of tritium-labeled estrone sulfate, estradiol-3-sulfate and estriol-16-glucosiduronate to the unhydrolyzed urine. The procedure provides for sufficient purification and thus virtually eliminates unspecific fluorescence as well as fluorescence-quenching impurities. This method proves to be accurate, precise and particularly applicable for lowtiter urines. One operator may complete 8 assays within one working day. Normal values for total estrogen excretion during the menstrual cycle, in amenorrhea, and after menopause are presented.
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