Use of an Organic Base in the Zimmermann Reaction*

Bongiovanni, Alfred M.; Eberlein, Walter R.; Thomas, P

doi:10.1210/jcem-17-2-331

Cited by 29 publications

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“…Blue tetrazolium (15) was utilized primarily for the measurement of steroids with the 16a-ol-17-one grouping, since these do not react with the Zimmermann reagent. The Zimmermann reaction and the measurement of total urinary 17-ketosteroids were conducted as reported elsewhere (16).…”

Section: Methodsmentioning

confidence: 99%

The ADRENOGENITAL SYNDROME WITH DEFICIENCY OF 3β-Hydroxysteroid DEHYDROGENASE*

Bongiovanni¹

1962

J. Clin. Invest.

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293

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A brief account of three cases of the adrenogenital syndrome due to a deficiency of 3,8-hydroxysteroid dehydrogenase has been published (1, 2). It is the purpose of this report to extend the original observations, detail the methodological aspects, and present certain clinical correlations.'The deficiency of 11,8-hydroxylase or 21-hydroxylase as two possible causes of the adrenogenital syndrome with adrenocortical hyperplasia is now well recognized. These aspects have been reviewed in detail recently (2). A direct demonstration of an enzymatic defect in adrenocortical tissue has been possible with respect to 21-hydroxylase in only two instances (3). Such experiments are not generally possible because of the obvious difficulties in obtaining suitable tissue from human subjects with this disorder. The evidence is generally indirect and depends upon the demonstration of "abnormal metabolites" in relatively large amount in the urine or blood. The nature of these metabolites provides the clue to the site of the defect, hence the indirect evidence for the deficient enzymatic transaction. A deficiency of the enzyme 3,/-hydroxysteroid dehydrogenase represents a third basis for the dis-* This work has been supported by Grants A-619 (C7) and 2A-5197 (C2) from the National Institute of Arthritis and Metabolic Diseases, U. S. Public Health Service.1 The following abbreviations have been employed for the steroids under discussion: dehydroepiandrosterone= 3,B-hydroxy-A5-androstene-17-one; androsterone = 3a-hydroxyandrostane-17-one; etiocholanolone = 3a-hydroxyetiocholane-17-one; pregnanetriol = pregnane-3a,17a,20a-triol; 11-ketopregnanetriol = 3a,17a,20a-trihydroxypregnane -11 -one; tetrahydro -E = 3a,17a,21 -trihydroxypregnane-11,20-dione; pregnenetriol = AY-pregnene-3fi,17a,20a-triol; pregnenediolone = 3,8,17a-dihydroxy-A'-pregnene-20-one; pregnenediol = A'-pregnene-3#,20a-diol; pregnenetetrol = ,17a,20,8,8, Six cases of this new form of the disorder are herein reported and compared with five cases due to a deficiency of 21-hydroxylase and five normal subjects. MATERIALS AND METHODSA summary of the clinical data on six cases of the adrenogenital syndrome due to a deficiency of 3#-hydroxysteroid dehydrogenase (I-VI), five due to a deficiency of 21-hydroxylase (VII-XI), and five normal individuals (XII-XVI) is presented in Table I. The post-mortem findings in four (I, III, IV, and V) were consistent with those of adrenocortical hyperplasia. The sex of the cases with the adrenal disorders was determined by examination of the sex chromatin in cells from the buccal mucosa, examination of the internal ducts by X ray, and histologic examination of the gonads in Case VI and in those who succumbed. Cases III and IV were known to have one prior affected sibling who died and showed bilateral adrenal hyperplasia at autopsy.Urine specimens were frozen immediately upon the completion of the 24-hour collection and stored at -100 C until analysis. The urinary steroidal compounds were first hydrolyzed with beta glucuronidase (Ketodase, War...

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Section: Methodsmentioning

confidence: 99%

The ADRENOGENITAL SYNDROME WITH DEFICIENCY OF 3β-Hydroxysteroid DEHYDROGENASE*

Bongiovanni¹

1962

J. Clin. Invest.

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293

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“…A procedure for the determination of ll-oxygenated, 17-ketogenic steroids in urine has been outlined by Wotiz et al (285) ; in this assay the steroids are isolated by paper chromatography and measured colorimetrically by the Zimmermann reaction. The technical advantage of using N-benzyltrimethyl ammonium methoxide rather than potassium hydroxide in the Zimmermann reaction has been de' scribed (286). A colorimetric method for the chemical determina tion of aldosterone has been recorded (287,288).…”

Section: Aoren Al Cortexmentioning

confidence: 99%

Endocrinology

Asper¹,

Wilson²

1958

Annu. Rev. Med.

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Progress in endocrinology has been rapid. So large is the field that in one year's harvest an entire chapter could be devoted to each of the endo crine glands. Remarkable achievements have been in many instances be cause of the successful application of newer techniques such as chromatog raphy, radioisotope tracer procedures, and microchemical analyses. In this review an appraisal has been made of experimental work relating to the anterior pituitary, thyroid, adrenal cortex, parathyroid, and pancreas. ANTERIOR PITUITARYMuch interest has been shown in the hormones of the anterior pituitary. Variability of experimental data pertaining to these hormones has resulted, in large measure, from the use of impure preparations and, although im proved methods of purification have become available, they remain expen sive and laborious. Highly purifi ed hormones are now obtainable for re search purposes from the Endocrinology Section of the National Institutes of Health (1), and it seems likely that'their use will lead to a better appraisal of the specifi c action of these substances.For many years attempts have been made to ascribe specific hormonal functions to the various cells of the pituitary. At first there was only con jecture, but later assignation of function to specific cell types was based on histological examination of the pituitary in patients with various glandular disorders. More recently, special stains and solvents, bioassays, and selective destruction of minute parts of the pituitary have been applied to this analy sis. Many such studies have been performed and the definitive results of a recent project are outlined by Barrnett et al.(2 ). These workers show that thyrotropic, follicle-stimulating, and luteinizing hormones are localized to those basophiles which have an affinity for periodic acid-Schiff stain.Gro wth ho rmone (GH}.-It has been recognized that growth hormone from bovine or porcine sources is ineffective in man. Indeed, a degree of 'The survey of literature pertaining to this review was completed in August, 1957.For the sake of brevity the abbreviation et ai, is used when three or more authors' names appear on a paper. S The following abbreviations are used: GH (growth hormone); ACTH (adrenocorticotropin); MSH (melanocyte-stimulating hor;mone); TSH (thyroid stimulating hormone); FSH (follicle-stimulating hormone); LH (luteinizing hormone) : PBI (protein-bound iodine) : BMR (basal metabolic rate): TBP (thy roxine-binding protein) : PTH (parathyroid hormone ) : GFR (glomerular filtration rate): Tmp (maximal rate of tubular reabsorption of phosphorus by kidney): Tmo (maximal rate of tubular reabsorption of glucose by kidney).

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“…The chromatograms were carried out on fluorescent silica gel-coated plates in chloroform-ether (9:1, v/v). After being examined under the ultraviolet lamp, the spots were sprayed with a modified Zimmermann reagent for the identification of keto steroids (Bongiovanni, Eberlein & Thomas, 1957) and with conc. H2SO4-ethanol (1:1, v/v), which stains most of the steroids characteristically.…”

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confidence: 99%

Hormonal steroids in the tissue of induced rat mammary tumours

Briziarelli¹,

Lusignani²,

Pedrali³

1969

Biochemical Journal

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The possible presence of steroids in the tissue of induced hormone-dependent rat mammary tumours was investigated. The method used involves a preliminary extraction of tumours followed by chemical separation and thin-layer chromatography. The identified compounds were cholesterol, androst-4-ene-3,17-dione, 5beta-androst-1-ene-3,17-dione, androsta-1,4-diene-3,17-dione and oestrone. This is the first report of the presence of these steroids in the tissue of an experimental tumour of a non-endocrine organ. In particular 5beta-androst-1-ene-3,17-dione has not previously been identified from natural sources.

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Use of an Organic Base in the Zimmermann Reaction*

Cited by 29 publications

References 0 publications

The ADRENOGENITAL SYNDROME WITH DEFICIENCY OF 3β-Hydroxysteroid DEHYDROGENASE*

The ADRENOGENITAL SYNDROME WITH DEFICIENCY OF 3β-Hydroxysteroid DEHYDROGENASE*

Endocrinology

Hormonal steroids in the tissue of induced rat mammary tumours

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