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Steroid monosulphate and disulphate fractions were obtained by chromatography on Sephadex LH-20 from human amniotic fluid collected at term. Only 16-hydroxydehydroepiandrosterone could be identified in the monosulphate fraction, the detection limit being 5–8 µg./1000 ml. amniotic fluid. In the disulphate fraction 5-androstene-3β,17-diol, 5-androstene-3β,17β-diol, 5-pregnene-3β,20-diol, 5-pregnane-3β,20-diol, 16-hydroxydehy-droepiandrosterone, 16β-hydroxydehydroepiandrosterone, 3β, 17β-dihy-droxy-5-androsten-16-one and 5-pregnene-3β,17,20-triol were identified by gas—liquid chromatography and gas chromatography—mass spectrometry. The concentrations of these steroids were determined in a pool of human amniotic fluid taken at term. The main compound was 5-androstene-3β,17-diol.