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
Ultracentrifugation and gel filtration were used to test for specific insulin-I-131 binding protein in the serum of normal subjects and in patients with overt or potential diabetes.
Such technics permit study of sera at physiologic pH, with physiologic concentrations of exogenous insulin added. They also eliminate the interference of electrical fields and, in ultracentrifugation, the need for artificial supporting media.
Whereas a specific binding protein was readily demonstrable in the serum of insulin-treated patients with high antibody titers, there was no evidence of a protein capable of binding exogenous labeled insulin in normal subjects nor in the serum of subjects with various types of diabetes and potential diabetes.
Serum from subjects who have a hyper-response of insulin to glucose loads, possibly caused by peripheral antagonism to insulin action, did not bind significant amounts of insulin-I-131.
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