CHROMATOGRAPHIC IDENTIFICATION OF ESTRIOL AND 16,17-EPIESTRIOL AS CONSTITUENTS OF THE URINE OF THE LAYING HEN

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Abstract
Presumptive estriol and 16,17-epiestriol fractions were separated from laying hens' urine by subjecting the hydrophilic fraction of the urinary phenolic extract to successive thin-layer chromatography (t.l.c), alumina-column chromatography, further t.l.c, and finally, microsublimation. The presence of estriol in the presumptive estriol fraction was confirmed by comparison of the thin-layer chromatographic behaviors of the phenol and four derivatives thereof with the behaviors of reference estriol and four corresponding derivatives of this reference material each in three different solvent systems. The presence of 16,17-epiestriol in the presumptive 16,17-epiestriol fraction was confirmed by similar comparisons of the presumptive 16,17-epiestriol fraction and five derivatives thereof with reference 16,17-epiestriol and five corresponding derivatives each in three solvent systems; and chromatographic correspondences for two further derivatives were observed in one solvent system each.It is concluded that estriol and 16,17-epiestriol are normal constituents of hens' urine.