Steroid conjugates were extracted from amniotic fluid samples with the non-ionic resin Amberlite XAD-7. After a two-step hydrolysis procedure two fractions of steroids, glucuronides, and sulfates and sulfoglucuronides, were obtained. The quantitation of the steroids was performed by glass capillary gas chromatography. The following concentrations (μg/l) of steroids were found in the glucuronide fraction of 20 amniotic fluid samples in uncomplicated pregnancies at term: mean (range) pregnanediol 183 (41-369), pregnanolone 43 (12-81), 32α,6α-dihydroxy-5β-pregnan-20-one 48 (20-135), 3α,16α-dihydroxy-5α-pregnan-20-one 161 (50-391), pregnanetriol 60 (29-119), and estriol 616 (117-1713). Concentrations of steroids in the fraction containing sulfates and sulfoglucuronides were: 16α-hydroxypregnenolone 52 (20-165), 16α-hydroxyDHA 165 (14-781), 3β,17β-dihydroxy-5-androsten-16-one 53 (15-120). and estriol 218 (51-485). In two cases of anencephaly concentrations of pregnanediol and other progesterone metabolites were within the normal range but those of estriol precursors and estriol were low or undetectable. © 1979.