Isolation of Androsterone Sulfate

It has long been recognized that many of the steroids, including the androgens, are excreted in urine in conjugation with other substances which render them not only more water-soluble but also neutralize their physiological activity to a great extent. In 1936 Cohen and Marrian (1) isolated estriol glucuronide, the conjugated form of estriol, from the urine of pregnant women and in 1936 Venning and Browne (2) reported the isolation of pregnanediol glucuronide from the same source. Schachter and Marrian in 1938 (3) isolated estrone sulfate from the urine of pregnant mares and in this paper we report the isolation of a conjugated androgen, androsterone sulfate, from the urine of a man suffering from an interstitial cell tumor of the testis with general metastases. Since this finding was reported (4), Munson, Gallagher, and Koch (5) have recently isolated a .17-ketosteroid sulfate in the form of a semicarbazone from normal men’s urine which on hydrolysis yields dehydroisoandrosterone. The first indication that the androgens might be excreted as conjugates was suggested by the observations of Funk, Harrow, and Lejwa in 1929 (6) that extraction of highly acidified male urine gave a better yield of the hormone promoting comb growth than extraction of neutral urine, and later Adler in 1934 (7) found that a physiologically inactive extract could be obtained from male urine with butanol. By subsequent heating with trichloroacetic acid the inactive extract could be converted into a potent substance which promoted the growth of the comb of the capon. In the summer of 1941 through the courtesy of Professor P. Masson we were fortunate in being able to collect urine over several weeks from a patient with extensive metastases from an interstitial cell tumor of the testis. The history and the pathological findings of this case have been published by Professor Masson (8). In 1932 a tumor of the left testis was removed from a 32 year-old laborer. For 9 years the man continued to work and was in perfect health. In May, 1941, he consulted a doctor, because his abdomen was enlarging and he found it difficult to bend when digging at work. On examination it was found that an enormous mass filled the abdomen and there was a tumor of one rib. The patient showed no sign of cachexia or of unusual hair growth or overdevelopment of the secondary sex organs; his strength