DOUBLE ISOTOPE DILUTION DERIVATIVE TECHNIQUE FOR TESTOSTERONE GLUCURONOSIDE IN URINE

1968 ◽  
Vol 58 (1) ◽  
pp. 77-97 ◽  
Author(s):  
Maria I. New ◽  
Joel M. Gross ◽  
Ralph E. Peterson
Keyword(s):  
Isotope Dilution ◽  
Adrenal Glands ◽  
Control Mechanism ◽  
Biological Fluids ◽  
Adrenal Hyperplasia ◽  
Physiological Control ◽  
Idiopathic Hirsutism ◽  
Normal Males ◽  
Double Isotope

ABSTRACT A double isotope dilution derivative technique for the determination of testosterone glucuronoside in urine has been developed. This is a highly specific and sensitive method which is applicable to the measurement of testosterone in submicrogram amounts in urine and other biological fluids. This method has been used to study normal prepubertal and pubertal males, normal females and subjects with gonadal dysgenesis, Cushing's syndrome, congenital adrenal hyperplasia, idiopathic hirsutism, Addison's disease and hypogonadism. The patients were studied after stimulation and suppression of gonads or adrenal glands to delineate the source of the androgen and its physiological control mechanism. The testosterone production of normal males determined by the method presented herein corresponds to results of others.

A DOUBLE ISOTOPE DERIVATIVE METHOD FOR THE DETERMINATION OF ALDOSTERONE IN PERIPHERAL PLASMA

1961 ◽  
Vol 37 (4) ◽  
pp. 541-558 ◽  
Author(s):  
Ejgil Bojesen ◽  
Hans Degn
Keyword(s):  
Isotope Dilution ◽  
Half Life ◽  
Acid Anhydride ◽  
Hydroxyl Group ◽  
Aldosterone Level ◽  
Peripheral Plasma ◽  
Derivative Method ◽  
Double Isotope

ABSTRACT The details of a method for the determination of aldosterone at the level of 1 ng or more are described. A combination of the isotope derivative principle and the isotope dilution principle, using 35S and 131I labelled piodobenzenesulfonic acid anhydride (pipsan) as the labelled reagents, has been employed. A quantitative or almost quantitative and reproducible esterification of the 21-hydroxyl group of aldosterone and an 80% recovery of the steroid added to plasma can be achieved by the procedure described. The blank value has been determined by the analysis of epripheral plasma of adrenalectomized dogs and was found to be indistinguishable from zero and therefore less than 3 ng per 100 ml of plasma. In salt depleted anaesthetized dogs the aldosterone level was 20–30 ng per 100 ml. The concentration increased considerably during adrenalectomy and decreased with a »half life« of 20–30 minutes from the time this was completed.

THE DETERMINATION OF 5α-ANDROSTANE-3α,17β-DIOL IN HUMAN PLASMA BY RADIOIMMUNOASSAY

1978 ◽  
Vol 88 (4) ◽  
pp. 778-786 ◽  
Author(s):  
P. Laband ◽  
J. A. F. Tresguerres ◽  
B. P. Lisboa ◽  
U. Volkwein ◽  
J. Tamm
Keyword(s):  
Thin Layer ◽  
Thin Layer Chromatography ◽  
Human Plasma ◽  
Cross Reactions ◽  
Idiopathic Hirsutism ◽  
Coefficients Of Variation ◽  
The Cross ◽  
Normal Males ◽  
Layer Chromatography

ABSTRACT Antibodies have been raised in rabbits against 3α,17β-dihydroxy-5α-androstane-6-0-carboxymethyloxime coupled with Cohn's fraction IV-4. The antiserum exhibited significant cross reactions with 5β-androstane-3α,17β-diol, 5α-dihydrotestosterone, and testosterone. No cross reactions were observed with 5α-androstane-3β,17β-diol and 5-androstene-3β,17β-diol. The methodological criteria for the measurement of 5α-androstane-3α,17β-diol in human plasma were as follows: The specificity was ensured by separating the cross reacting steroids by thin layer chromatography. The intraassay and interassay coefficients of variation were found to be 6.2 and 10.2 %, respectively. The sensitivity was 30 pg. The recovery of different amounts of 5α-androstane-3α,17β-diol added to human plasma (80, 120, and 200 pg) yielded 91.3, 92.5, and 93.5%, respectively. The following concentrations of 5α-androstane-3α,17β-diol have been determined in human plasma (mean ± sd, ng/dl): Normal males: 18.98 ± 5.9; normal females: 2.65 ± 0.27; females with idiopathic hirsutism: 11.9 ± 6.4; pre-pubertal children: not detectable.

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