A micro-method has been developed for the determination of Δ4-3-ketosteroids and Δ5-3β-hydroxysteroids, oestradiol-17β, oestrone and oestriol in small volumes of cyst fluid. The method consists of separation of the neutral and phenolic fractions by solvent partition; fractionation of the neutral steroids by partition and paper chromatography and of the three oestrogens by adsorption chromatography after methylation. This technique has been applied to cyst fluid from the ovaries of twenty-one patients, twelve with the Stein—Leventhal syndrome, and nine with other gynaecological disorders. In the patients with the Stein—Leventhal syndrome the steroid concentrations in cyst fluid showed a definite pattern characterized by high concentrations of Δ4-androstenedione, low concentrations of progesterone and 17α-hydroxyprogesterone, and small amounts or undetectable oestrogens. In cyst fluid from the ovaries of the nine control patients there were, in general, lower concentrations of Δ4-androstenedione and significant amounts of oestradiol-17β. High concentrations of progesterone and oestrogens were found in the cyst fluid of patients with menorrhagia and endometriosis.
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