[Rapid measurement of estriol & estradiol by high performance liquid chromatography (HPLC)]

Measurement of Estriol (E3) and Estradiol (E2) within 25 minutes by high performance liquid chromatography (HPLC) was achieved in this study and the values were compared with those of radioimmunoassay (RIA). A totally computerized HPLC method was developed for measuring unconjugated E3 and E2 in the sera of pregnant women. The sera samples were injected directly into the apparatus and transferred to a pretreatment column where estrogens were absorbed while hydrophilic components such as proteins and carbohydrates were excluded. The estrogens then passed through another separation column containing a new type of polymer gel. The mobile phase consisted of an acetonitrile/water mixture, and separation was achieved by means of a reversed phase mechanism. The eluent was monitored for fluorescence. All these procedures were monitored and controlled with a built-in microcomputer. The sera samples from 61 normal pregnant women at 20 to 41 weeks of pregnancy were simultaneously assayed by HPLC and RIA. The correlation of values obtained from HPLC and RIA was; E3: Y = 0.875X-0.172, the coefficient of correlation was 0.899, E2: Y = 0.972X + 6.791, the coefficient of correlation was 0.841 (Y = RIA values, X = HPLC values). The quick measurements of E3 and E2 by HPLC can be a useful method in evaluating the feto-placental function.