Determination of Urinary Testosterone by Chromatography and Colorimetry: Findings in Normal Subjects and in Patients with Endocrine Diseases1

In this method for measuring conjugated urinary testosterone, testosterone-4-14C is added to an aliquot of urine to correct for subsequent losses and an ethyl ether extract is then chromatographed in 2 Zaffaroni systems prior to chromic oxide oxidation of testosterone to Δ4-androstene-3,17-dione. A third Zaffaroni system confirms the completion of oxidation and purifies the steroid before a final fourth chromatography in a Bush system. Quantitation is carried out by a modified micro Zimmermann reaction. Values of testosterone glucuronide in 24-hr specimens (mean ±se) were found to be 19 ±3 μg in 20 normal women aged 20–40; 88±7 μg in 20 normal men aged 30–40; 151± 22 μg in 5 young normal men aged 17–24; and 6 ±3 μg in 5 boys aged 7–12. A satisfactory correlation between the level of testosterone glucuronide and the degree of androgenicity in various pathologic states studied was observed.