Clinical Method for the Chromatographic-Colorimetric Determination of Urinary 17-Ketosteroids. I.

Following the demonstration by Wilkins, Lewis, Klein, and Rosenberg (1) that cortisone controls the clinical manifestations and abnormal laboratory findings in virilizing adrenal hyperplasia, Bartter, Albright, Forbes, Leaf, Dempsey, and Carroll (2) postulated a deficient synthesis of "sugar hormone" (17-hydroxycorticosterone, Compound F) in this disease, reasoning from the unusual response of such patients to the administration of adrenocorticotropin (ACTH). This hypothesis has been supported by the demonstration of low blood levels of 17-hydroxylated-corticosteroids ("Porter-Silber chromogens") in untreated patients with adrenal hyperplasia and the usual failure of these levels to rise, following administration of ACTH (3,4). The discovery that, routinely, large quantities of pregnane-3a, 17a,20a-triol are excreted in the urine of patients with virilizing adrenal hyperplasia (5) has suggested a possible site of the metabolic block in the synthesis of Compound F.The present studies were undertaken to characterize the corticosteroids excreted in the urine in adrenal hyperplasia, in an attempt to define more precisely the specific defect in steroid biosynthesis. METHODSTwenty-four-hour collections of urine from normal control subjects and from patients with adrenal hyperplasia were investigated in the following manner:Urine for 17-ketosteroid determinations was hydrolyzed first with beta-glucuronidase2 and manually extracted with ether; the remaining urine was continuously extracted at room temperature with ether for 36 hours after the addition of 10 volumes per cent (v/v) of 40 per cent sulfuric acid. The combined extracts were fractionated on alumina columns (6) tions were submitted to infra-red spectrophotometric analysis.The level of urinary neutral 17-ketosteroids produced by bismuthate oxidation ("17-ketogenic steroids") was determined according to the method of Brooks and Norymberski (7).The quantities of pregnane-3a,17a,20ca-triol were determined in urine hydrolyzed with beta glucuronidase2 and chromatographed on alumina according to the method of one of the authors (5).Crude extracts of urine hydrolyzed with beta glucuronidase were chromatographed on florisil (8). The fraction eluted with 25 per cent methanol in chloroform was examined by the following techniques: a) The phenylhydrazine-sulfuric acid method of Porter and Silber (9); b) periodate oxidation for the simultaneous determination of formaldehydogenic and acetaldehydogenic steroids (10); c) phosphomolybdate method of Heard and Sobel (11); d) blue tetrazolium method of Chen, Wheeler, and Tewell (12); e) 2,4-dinitrophenylhydrazine method of Gornall and Macdonald (13).Urines collected from patients during therapy with cortisone were obtained two weeks to six months following the introduction of the intramuscular administration of Compound E acetate in doses ranging from 25 to 75 mg. given every third day. In each case, the specimen studied was obtained at a time when the total urinary 17-ketosteroids were significantly depressed (1). Each of the m...

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“…The combined extracts were fractionated on alumina columns (6) and suitable frac-tions were submitted to infra-red spectrophotometric analysis.…”

Section: Methodsmentioning

confidence: 99%

Partial Characterization of Urinary Adrenocortical Steroids in Adrenal Hyperplasia 1

Eberlein¹,

Bongiovanni²

1955

J. Clin. Invest.

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“…The remaining fractions, two ketonic and two non-ketonic (3a and 3/3 alcohols), were analysed by chromatographic methods. The ketonic fraction was followed by a modification of the Dingemanse technique (Dingemanse, Huis in't Veld, and de Laat, 1946); the non-ketonic fraction by conversion of the component alcohols to 3 5 dinitrobenzoates (Kellie, Smith, and Wade, 1953;Kellie and Wade, 1953).…”

Section: Methodsmentioning

confidence: 99%

Urinary Steroid Excretion in Rheumatoid Arthritis

Kellie¹,

Wade²

1953

BMJ

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“…Urinary total 17-ketosteroids [method by Vestergaard, 1951: 5-17 Urinary total 17-ketosteroids [Vestergaard, 1951: 5-17 mg/24 h]: 12.27 mg. Adrenal fractions of urinary total 17-ketosteroids (Dingemaanse et al, 1946, Adezati el al., 1957: dehydroepiandrosterone (0.7-2.5 mg/24 h): 0.235 mg; 1 l-hydroxy-17-ketosteroids (0.6-1.3 mg/24 h): 0.109 mg.…”

Section: Endocrinological Investigationsmentioning

confidence: 99%

Addison-Schilder’s Disease Report of a Case with Probable Relapsing Addisonian Encephalopathy

Bugiani¹,

Balestreri²

1971

Stereotact Funct Neurosurg

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Clinical Method for the Chromatographic-Colorimetric Determination of Urinary 17-Ketosteroids. I.

Cited by 78 publications

References 0 publications

Partial Characterization of Urinary Adrenocortical Steroids in Adrenal Hyperplasia 1

Partial Characterization of Urinary Adrenocortical Steroids in Adrenal Hyperplasia 1

Urinary Steroid Excretion in Rheumatoid Arthritis

Addison-Schilder’s Disease Report of a Case with Probable Relapsing Addisonian Encephalopathy

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