Three highly polar unconjugated Δ4-3-oxosteroids containing a 6β-hydroxyl group have been isolated from human pregnancy urine. They have been identified as 6β,17α,20β,21-tetrahydroxypregn-4-ene-3,11-dione (compound 3), 6β,11β,17α,20α,21-pentahydroxypregn-4-en-3-one (Compound 4c) and 6β,11β,17α,20β,21-pentahydroxypregn-4-en-3-one (Compound 4d). Two further steroidal compounds were found, but have not as yet been identified. The isolation procedure involved the use of thin-layer chromatography for purification of the crude urinary extracts and paper chromatography for the isolation of the individual steroids. Chemical transformations and spectroscopic analyses have been used for the identification of the individual compounds.