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Free and conjugated oestrogens were determined in the fluid from ovarian cysts of normally menstruating women, of subjects under gonadotrophin stimulation and of patients with different gynaecological conditions. The fluid was first extracted with ether to yield free oestrogens, and then the aqueous residue was submitted to acid hydrolysis. Three samples of ovarian venous blood were assayed by the same procedure. Indirect evidence that the oestrogens measured after acid hydrolysis were originally present as chemical conjugates was obtained.

The results indicated that conjugated oestrogens were synthesized in the ovary. The amount of oestrogens in cyst fluid varied with the phase of the cycle, and was increased in patients treated with gonadotrophins, in subjects with high secretion of gonadotrophins, and in some patients with functional uterine haemorrhage. It was low in patients with the Stein-Leventhal syndrome. Conjugate concentration was low relative to the concentration of free oestrogens in fluid from follicles and cysts derived from follicles, and similar to that of free oestrogens in corpora lutea, corpus luteum cysts and in patients with irregular bleeding. The concentration of conjugated oestradiol appeared to be generally independent from that of conjugated oestrone.

The relative proportions of free and conjugated oestrogens were different in cyst fluid and in ovarian venous blood.