IN an earlier publication (Robinson, 1948) the results obtained in a study of 17-ketosteroid excretion in men of different age-groups were reported. Special attention was paid to quantitative dlifferences in total ketosteroid excretion during the later decades of life. The primary object of the work was to form a basis for comparison with similar quantitative data from cases of prostatic cancer.In parallel with this quantitative study a qualitative examination of these urines was also undertaken. This involved separation and estimation of the individual components of the total ketosteroid complex present in urinary extracts. Such a separation can be achieved by chromatographic methods, and many investigators have described procedures (e.g. Callow, 1939; Wolfe, Fieser and Friedgood, 1941; Dobriner, Rhoads, Lieberman, Hill and Fieser, 1944; Dobriner, Lieberman and Rhoads, 1948). In most of the work described by these authors the components were recognized by isolation and identification by the usual chemical criteria such as melting-point and optical rotation. Comparatively large quantities of material are required for identification by isolation, and the method is unsuitable for the amounts obtained from small specimens of urine.Dingemanse, Huis in't Veld and de Laat (1946) described a method for chromatographic-colorimetric determination of urinary 17-ketosteroids which is applicable to the amounts encountered in one day's output of urine. The method involves adsorption of the 17-ketosteroids on a column of alumina from benzene solution followed by fractional elution with benzene of increasing ethanol content. The amount of 17-ketosteroid present in each eluate is determined by the Zimmermann reaction. Preliminary experiments with the method described by these authors showed that separation and estimation of the steroid components could be achieved, but that reproducible results depended on careful standardization of the alumina and of the conditions of elution. With these precautions this method was adopted in the present work.~-1~~METHODS AND MATRIALS.(1) Preparation of urine extract8.Hydrolysis and extraction of the urines were carried out by the same method as that given in the earlier paper (Robinson, 1948). For the chromatograms 5 mg. of total 17-ketosteroid was found to be the most suitable amount, and sufficient urine from a 24 or 48-hour collection was hydrolysed and extracted to furnish a quantity in excess of 5 mg. The required volume of urine was estimated approximately by a preliminary assay of total 17-ketosteroid on a small aliquot

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