HYDROLYSIS OF CONJUGATED ESTROGEN FRACTIONS IN HUMAN PREGNANCY URINE

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.