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

Abstract

In this method for measuring conjugated urinary testosterone, testosterone-4-14 C 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 A 4 -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 ng in 20 normal women aged 20-40; 88+7 ng in 20 normal men aged 30-40; 151 ±22 ng in 5 young normal men aged 17-24; and 6 ±3 jug 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. (J Clin Endocr 25: 95, 1965) T ESTOSTERONE 5 is properly considered the most potent of the known androgens. The growing evidence that low or normal values of 17-ketosteroids (17-KS) may coexist with virilization in the female has made the evaluation of testosterone indispensable to an adequate investigation of androgenic states.Free testosterone has been measured in blood and a good correlation was established between elevated values and virilizing conditions in women (1, 2). Testosterone glucuronide has been measured in human urine (3-6). A new, relatively simple method is described in this paper for the measurement of conjugated

Androstenedione and Its Conversion to Plasma Testosterone in Congenital Adrenal Hyperplasia*

“…Since testosterone is the most potent natural androgenic hormone and the adrenal is capable of synthesizing testosterone (4), increased secretion of this steroid has been suggested as the responsible mechanism. This hypothesis is supported by the demonstration of increased urinary excretion (5)(6)(7)(8)(9)(10)(11), plasma concentration (10,(12)(13)(14), and production rate (10)(11)(12)(13)(14)(15)(16)(17)(18)(19) of testosterone in patients with congenital adrenal hyperplasia. However, recent work by Korenman and Lipsett (15) and Horton and Tait (16) indicates that the weak androgen, androstenedione (androst-4-ene-3, 17- ther demonstrated that almost two-thirds of blood testosterone in the normal female is derived from blood androstenedione.…”

Androgen metabolism in man. Some current concepts.

“…Normal values for the excretion of testosterone glucuronide have been obtained in Addison's disease (Ibayashi et al, 1964;Vermuelen, 1966), and consistently raised values of excretion and plasma-testosterone level and production have been found in congenital adrenal hyperplasia (Burger et al, 1964;Futterweit et al, 1964;Casey, 1965 ;Korenman et al, 1965;Lim and Dingman, 1965 ;Rosner et al, 1965;Degenhart et at., 1966) but the correlation with 17-oxosteroid excretion is not close; there is in this condition good suppression of testosterone with corticosteroid. Few observations are available in Gushing's syndrome with adrenal hyperplasia, but urinary testosterone glucuronide is increased (Ibayashi et al, 1964Vermuelen, 1966) and a high level of testosterone has been observed in the adrenal vein (Hudson et at., 1963) and a low level in the peripheral plasma of such a patient after adrenalectomy .…”

Excretion and Production of Testosterone in Normal Children, in Children with Congenital Adrenal Hyperplasia, and in Children with Precocious Puberty

“…A few series of patients with CAH have been published ; rates of production and excretion of testosterone were determined and were found to be elevated [25,36]. KORENMAN et al [21] describe a case with only a slightly increased testosterone UPR and one case with an elevated UPR.…”