The Plasma Cortisol Response to Acth in “Idiopathic Hirsutism”*

Abstract

The biochemical abnormalities in patients with "idiopathic hirsutism" are imperfectly understood. The fractionation and measurement of urinary 17-ketosteroids in these patients have been studied in several laboratories. It appears that this disorder is associated with increased production of an adrenal androgen which is reflected by increased excretion of urinary C1902 17-ketosteroids, in particular, androsterone and etiocholanolonel (1-7). Gallagher and co-workers (1), Kappas, Pearson, West and Gallagher (2) and Brooks and Prunty (3) reported that the urinary 11-oxy-17-ketosteroids may also be increased in these patients, and Bush and Mahesh (4) noted, in particular, increased excretion of 1 1-hydroxyandrosterone. These 11-oxygenated steroids are presumed to be derived principally from cortisol and 11-hydroxyandrostenedione metabolism (8). By inference cortisol production in patients with idiopathic hirsuitism has been thought to be either normal or high. To define further the nature of this adrenal abnormality, plasma cortisol levels have been studied in hirsute patients during 8 hours of intravenous ACTH stimulation.The determination of plasma 17-hydroxycorticoids has received wide acceptance and application in studies of abnormal adrenal function. * This study was supported by Grant C-1003 from the National Cancer Institute.t Postdoctoral Research Fellow, National Heart Institute.t Trainee, National Institute of Arthritis and Metabolic Diseases.1 In this paper the trivial names of the steroids have been used. Cortisol: 11,p,17a,21-trihydroxy-4-pregnene-3,20-dione. Prednisone: 17a,21-dihydroxy-1,4-pregnene-3,11,20-trione. Cortisone: 17a,21-dihydroxy-4-pregnene-3,11,20-trione. Etiocholanolone: 3a-hydroxy-etiocholane-17-one. Androsterone: 3a-hydroxy-androstane-17-one.11 -Hydroxyandrosterone: 3a,l 1 p-dihydroxy-androstane-17-one. 11 -Hydroxy-androstenedione: 11 f3-hydroxy-4-androstene-3, 1 7-dione. Pregnanetriol: 5p-pregnane-3a,17a, 20a-triol. Pregnanediol: 5,8-pregnane-3a,20a-diol.These methods generally depend upon the specificity of the Porter-Silber (9) reaction after partial purification of a chloroform or dichloromethane extract of plasma. While it appears that these methods satisfactorily reflect cortisol content of plasma, work by Bondy and co-workers (10) and Morris and Williams (11), in which the plasma extract was carried through more rigorous purification procedures before final quantitation, have given blood cortisol levels which were significantly lower than those obtained by the less specific procedures. Since "idiopathic hirsutism" has been postulated to represent a partial enzymatic block in cortisol synthesis (2, 5), similar to the enzymatic blocks now recognized in the congenital adrenal hyperplasias (12-14), it was felt desirable to measure blood cortisol in these patients. A paper chromatographic step to separate plasma cortisol from other adrenal products and their derivatives was used. A reliable, technically feasible method has been devised. MATERIALS AND METHODSControl subjects. Control ...

Increased Unbound Cortisol in the Plasma of Estrogen-treated Subjects*

“…The last plasma was divided into two equal aliquots, and the eight plasma samples were carried through the extraction procedure and paper chromatographic separation of cortisol previously described (17). The cortisol area from the paper chromatogram of the first and of the last samples was eluted and analyzed with the Porter-Silber reagent (18).…”

Suppression of the Pituitary-ACTH Response In Man By Administration of ACTH or Cortisol*†

“…Determination of plasma cortisol and substance S. These steroids were determined by a modification of a method which we have described previously for plasma cortisol (12). A 10-15 ml sample of plasma was brought to pH 9 with a lithium hydroxide solution, and then subjected to extraction three times with freshly distilled chloroform.…”

“…Cortisol and substance S were separated on a standard strip carried through the procedure, and were located with ultraviolet light. Equivalent areas were cut from the plasma chromatogram, and these areas were extracted with 3 methanol soaks, as previously described (12). Cortisol and substance S were measured quantitatively by the Silber-Porter method (14).…”

The in Vitro Red Blood Cell Uptake of C14-Cortisol; Studies of Plasma Protein Binding of Cortisol in Normal and Abnormal States*

“…Approximately 2.25 ml of plasma was cleanly separated from the packed cells, and an aliquot of the separated plasma (usually 2 ml, on rare occasions slightly less) was then pipetted into a 90-ml centrifuge tube. Ten ml of distilled water was added, and the pH was adjusted to 9.0 with a dilute lithium hydroxide solution [cortisol recovery appears to be more reproducible when lithium hydroxide is used for alkalinization (6) ]. The dilute alkaline plasma was extracted three times with 1.5 vol of freshly redistilled dichloromethane.…”

“…Initial plasma cortisol levels were determined by the method of Plager, Cushman and Chase (6) in which cortisol is separated by paper chromatography. Since a relatively large amount of cortisol was added to each blood sample, the final plasma cortisol concentration was calculated by the following equation: 3 Final plasma cortisol concentration = (initial plasma cortisol concentration + added cortisol concentration) X (100-% RBC uptake)…”