A rapid routine method of determining estrogens in non-pregnancy urine is described. It involves the following steps: 1) Dilution of each urine sample with an equal volume of saturated ammonium sulfate solution; 2) Addition of Sephadex G-15 beads; 3) Filtration of the mixture; 4) Liberation of the estrogens from the Sephadex by washing the beads on the filter with 0.1N NaOH; 5) Automated fluorimetric analysis of the alkaline filtrate according to the procedure used for the determination of estrogen in pregnancy urine. A correction for non-specific fluorescence remains necessary and is achieved by measuring background fluorescence with a second fluorimeter.