Urinary 17-Hydroxycorticosteroids Measured by Gas-Liquid Chromatography

Murphy, Denis; West, H. F.

doi:10.1677/joe.0.0360331

Cited by 19 publications

(2 citation statements)

References 11 publications

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“…Details of the routine assay procedure for urinary 17-hydroxycorticosteroids (17-OHCS) have been published (Murphy and West, 1966). In brief, 10 ml of urine (or much less, in special circumstances, made up to 10 ml with distilled water) is adjusted to pH 7 and 1 ml of potassium borohydride solution (15% in 0-1 N NaOH) added.…”

Section: Methodsmentioning

confidence: 99%

Routine multipurpose gas chromatographic assay for urinary corticosteroids

Murphy¹,

West²

1968

J Clin Pathol

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SYNOPSIS The use of a routine assay for urinary steroids is described that is thought to satisfy the following needs: the measurement of low levels of 17-hydroxycorticosteroid excretion; the assessment of adrenocortical activity during prednisolone therapy; the diagnosis of the adrenogenital syndrome and the monitoring of its treatment using any available urine specimen; the very early diagnosis of pregnancy using any available urine specimen. It may also prove of value in the assessment of progesterone secretion and metabolism during pregnancy.Corticosteroid assays are used for the measurement of hypo-and hyperfunction of the adrenal cortex; the assessment of iatrogenic adrenal suppression; the measurement of the degree of adrenal stimulation induced with ACTH; the diagnosis of the adrenogenital syndrome and the assessment of its treatment. If a single quick routine assay could provide unequivocal information for each of these needs and could provide, in addition, an early dignosis of pregnancy and an assessment of the excretion of a number of pregnancy steroids it would be of considerable use. This paper describes the clinical use of an assay that appears to provide such information. METHODSDetails of the routine assay procedure for urinary 17-hydroxycorticosteroids (17-OHCS) have been published (Murphy and West, 1966). In brief, 10 ml of urine (or much less, in special circumstances, made up to 10 ml with distilled water) is adjusted to pH 7 and 1 ml of potassium borohydride solution (15% in 0-1 N NaOH) added. After heating at 60°C for 50 min 1 ml of 25% acetic acid is added followed, in 2 min, by 4 ml of sodium metaperiodate solution (10% aqueous) and the heating continued for a further 10 minutes. One ml of 5 N NaOH is then added and the products extracted with 20 ml of ethylene dichloride. The extract is filtered through sodium sulphate and evaporated to dryness before acetylation at 60°C for 30 min with 0-2 ml of pyridine and 0-1 ml of acetic anhydride. The reagents are then evaporated under nitrogen and the residue dissolved in 0-4 ml ethanol for gas chromatography. When it has been necessary to obtain more information by forming trimethylsilyl ether (TMSi) derivatives, of the C,3 and C,17 hydroxyl functions, this was effected as follows. The dry unacetylated Received for publication 25 October 1967. extracts were reacted with 0-2 ml of purified pyridine and 0-1 ml of hexamethyldisilazane overnight at room temperature. The reagents were then evaporated under nitrogen and the residue was dissolved in 04 ml of ethylene dichloride ready for gas chromatography. The acetates were gas chromatographed at 235°C and the TSMi ether derivatives at 210°C. In the routine 17-OHCS assay the major C,21 metabolites of cortisol, ie, the glucosiduronates of tetrahydrocortisol, tetrahydrocortisone, the cortols, the cortolones and their 5a isomers are converted to 11 ,-hydroxyactiocholanolone and lI1p-hydroxyandrosterone and these are measured together in a single gas chromatograph peak. Pregnanetriol is converted to aeti...

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Section: Methodsmentioning

confidence: 99%

Routine multipurpose gas chromatographic assay for urinary corticosteroids

Murphy¹,

West²

1968

J Clin Pathol

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“…Gas liquid chromatography for the group separation of 17-0HCS was described by Murphy andWest in 1966 andin 1970 Makin examined various derivatives for the separation of adrenal steroids using a column of SE·30. He found formate esters suitable and in 1971 published details of a qualitative profile of 17-oxosteroids and 17-0HCS as an aid in the diagnosis of adrenal pituitary disorders.…”

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Group Determination of Urinary Adrenal Steroids by Gas Chromatography

Craig

Chamberlain

1972

Ann Clin Biochem

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Routine colorimetric methods for the determination of urinary metabolites of the adrenal cortcx tend to be unspecific. The separation of 17-hydroxycorticosteroids (l7-0HCS) into H-oxy and II-dcoxy fractions has usually entailed the use of column chromatography (Morris, 1959) or differential solvent extraction (Few, 1968).Gas liquid chromatography for the group separation of 17-0HCS was described by Murphy and West in 1966 and in 1970 Makin examined various derivatives for the separation of adrenal steroids using a column of SE·30. He found formate esters suitable and in 1971 published details of a qualitative profile of 17-oxosteroids and 17-0HCS as an aid in the diagnosis of adrenal pituitary disorders.We have used acetate csters for the quantitative assessment of lloOxy and ll-deoxy 17-0HCS and some of our experiences arc described in this paper.

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The Hydrolysis of Steroid Conjugates

Bradlow¹

1970

Chemical and Biological Aspects of Steroid Conjugation

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Urinary 17-Hydroxycorticosteroids Measured by Gas-Liquid Chromatography

Cited by 19 publications

References 11 publications

Routine multipurpose gas chromatographic assay for urinary corticosteroids

Routine multipurpose gas chromatographic assay for urinary corticosteroids

Group Determination of Urinary Adrenal Steroids by Gas Chromatography

The Hydrolysis of Steroid Conjugates

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