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So as to evaluate metabolism of catecholestrogens during pregnancy, changes in plasma unconjugated estradiol (E2), estrone (E1), 2-methoxyestradiol (2-mE2) and 2-methoxyestrone (2-mE1) concentrations were measured after a bolus injection of 40mg of estrone-sulfate (E1-S) or 100mg of dehydroepiandrosterone-sulfate (DHA-S) to normal pregnant women near term. The injection of E1-S resulted in a marked rise in plasma 2-mE1 levels followed by a rapid marked increase in the plasma E1 concentration. The injection of DHA-S produced a small increase in the plasma 2-mE2 levels in spite of a marked increase in the plasma E2 concentration. No significant correlation between E2 and 2-mE2 was found, however, significant correlations between E1 and 2-mE1, between E1 and 2-mE2 and between 2-mE2 in in both groups were demonstrated. The marked increase in 2-mE1 compared to a small increase in 2-mE2 after DHA-S injection indicates that E2 may participate in the formation of catecholestrogen via its conversion to E1 during pregnancy. These results suggest that the main route for the formation of catecholestrogen is via E2----E1----(2OHE1)----2-mE1 in normal pregnancy near term.

Recently it has been attached importance to the physiological function of catecholestrogens. To elucidate it a specific radioimmunoassay of conjugated 2-hydroxyestrone (2-OHE1) and 2-hydroxyestradiol-17 beta (2-OHE2) in human plasma was attempted. After extracting with ethyl acetate, samples were purified by a short Sephadex LH-20 columnchromatography and determined using the antiserum to 2-OHE1-17(O-carboxymethyl) oxime-bovine serum albumin conjugate. The following results were obtained: 1) A long Sephadex LH-20 columnchromatography which was used for the purification of catecholestrogens produced chemically, had a superior faculty in separation and high capacity 2) The antiserum cross-reacted 26.4% with 1-OHE2, but less than 1% with other steroid hormones. 3) The sensitivity of the method was around 10pg in both 2-OHE1 and 2-OHE2 assays. The method blank determined using 2ml plasma of bilaterally adreno-oophrectomized women was below 15 pg/ml in both 2-OHE1 and 2-OHE2 assays (n = 24). 4) The coefficient of variation in both accuracy and between-assay precision of the method was less than 17%. 5) The plasma 2-OHE1 concentration was below 15 pg/ml in normal men (n = 8) and non-pregnant women (n = 13). The concentration in pregnant women was 20 +/- 8 pg/ml (SD, n = 7) in 1st trimester of pregnancy, 58 +/- 13 (n = 3) in 2nd trimester and 177 +/- 66 (n = 12) in 3rd trimester. The E1/2-OHE1 and E2/2-OHE1 ratios in 3rd trimester of 9 pregnant women were 19.7 +/- 11.9(SD) and 78.0 +/- 27.8, respectively. 6) The plasma 2-OHE2 concentration was below 15 pg/ml in normal men (n = 8), non-pregnant women (n = 8), and 1st to 2nd trimester (n = 10) and 15 +/- 9 pg/ml (SD, n = 13) in 3rd trimester of pregnant women.