Determination of urinary placental estriol by reversed-phase liquid chromatography with fluorescence detection

Ww describe a liquid-chromatographic procedure for determining urinary estriol concentrations. The urine sample, after enzymatic hydrolysis to free the conjugated estrogen, is extracted with ether, and an aliquot of the resulting extraction residue is injected into the liquid chromatograph. Sample components are separated with a reversed-phase C18 column and isocratic elution with an acetonitrile/water mobile phase. Using a far-ultraviolet excitation wavelength, we measure the natural fluorescence of the eluted estrogen with a fluorescence detector. The procedure provides excellent sensitivity for determing near-term pregnancy concentrations of urinary estriol. The selectivity of the method limits the effect of potentially interfering compounds.