HYDROLYSIS OF CONJUGATED ESTROGEN FRACTIONS IN HUMAN PREGNANCY URINE

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Abstract
The release of six estrogen fractions from conjugation in human pregnancy urines has been studied using various hydrolytic methods. The estrogens concerned were estrone, estradiol-17β (estradiol), 2-methoxyestrone, 16-epiestriol, and a ring D ketolic fraction (mainly 16α-hydroxyestrone). Considerable amounts of urinary estrone and ring D ketolic estrogens may be conjugated in a non-glucuronide form. In these cases an enzyme preparation containing β-glucuronidase and sulphatase, prepared from the digestive juice of the snail Helix pomatia, proved to be superior to β-glucuronidase enzymes of bacterial or mammalian liver origin. Conventional hot acid hydrolysis yielded levels of estrone, estradiol, estriol, and 16-epiestriol which agreed fairly well with those obtained following snail enzyme hydrolysis. In some urines, hot acid treatment was not suitable for hydrolysis of conjugated 2-methoxyestrone. Optimum hydrolytic conditions for both normal and diabetic pregnancy urines were realized by incubating for 24 hours with 500 units of the snail β-glucuronidase and 250 units of sulphatase/ml of urine at pH 5.2 and 37–38 °C.