1. Pregnant cow's urine was hydrolysed with acid, and the lipid material obtained was submitted to the fractionation procedures customary in the study of urinary steroids.
2. Oestrone (0·3 mg/1.) and oestradiol-17α (0·1 mg/1.) were isolated from the phenolic fraction. Oestradiol-17β could not be detected.
3. The heterocyclic phenol equol isoflavan-7:4′-diol) was also isolated (6 mg/1.).
4. The neutral non-ketonic fraction contained 5β-androstane-3α: 17α-diol (0·2 mg/1.) and 5α-androstane-3β:17α-diol. A very small quantity of material resembling 5β-pregnane-3α:20α-diol (the common 'pregnanediol' of human pregnancy urine) was isolated but not satisfactorily identified.
5. The volatile part of this fraction contained two tetrahydroionanediols A and B.
6. The neutral non-volatile ketonic fraction was small (0·6 mg/1.). Three α-17-oxosteroids were tentatively identified.
7. The results are discussed in relation to the routes of steroid excretion in cows, and in relation to the determination of steroids in animal urines.
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