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A method is described for the determination of urinary 17-hydroxycorticosteroids using fluorescence high-performance liquid chromatography. After enzymatic hydrolysis, 17-hydroxycorticosteroids were extracted using an Extrelut column and then labeled with Dns-hydrazine in hydrochloric acid-ethanol solution. The labeled steroids were chromatographed on a microparticulate silica gel column, the mobile phase was dichloromethane-ethanol-water (900:60:40). The eluate was monitored on a fluorophotometer at 365 nm (excitation) and 505 nm (emission). Linearity of the fluorescence intensities (peak heights) of various 17-hydroxycorticosteroids were obtained between 60 pg and 20 ng. The assay was sensitive, precise and accurate. Comparison with the results obtained by radioimmunoassay gave correlation coefficients of 0.932 for tetrahydrocortisol and 0.930 for tetrahydrocortisone. The proposed method is clinically useful for the routine analysis of urinary 17-hydroxycorticosteroids.
The effects of levodopa on the plasma concentration of 11-hydroxycorticosteroids and glucose were examined in mice. In fasted, nialamide treated mice, but not in fed mice, levodopa produced a significant decrease in the plasma concentration of 11-hydroxycorticosteroids. This was accompanied by a significant and marked decrease in the plasma glucose concentration. These effects of levodopa could be mimicked by relatively small doses of dopamine injected intracerebroventricularly but not intravenously. The integrity of the hypothalamic-pituitary adrenal axis in nialamide-treated mice was suggested by the elevation in plasma 11-hydroxycorticosteroids produced by fasting or by insulin induced hypoglycaemia. These results indicate that in the mouse, as in other species, levodopa can inhibit stress provoked increases in the secretion of 11-hydroxycorticosteroids.
The 24-hour excretion of 17-ketosteroids and 17-hydroxycorticosteroids has been estimated in a series of male duodenal ulcer subjects and compared with that of 56 normal male controls. It has been found that both 17-ketosteroid and 17-hydroxycorticosteroid excretion is less in ulcer subjects than in the control group; these differences are not large but in the case of 17-hydroxycorticosteroids they are statistically significant. For active ulcers (107 men) 17-hydroxycorticosteroid excretion is approximately 78% of normal and 17-ketosteroid excretion 93% of normal; in the quiescent phase (50 men) the differences are rather larger, being respectively 71% and 86% of normal. This reduced excretion persists after operation in both the short term, six months after operation (53 men), and the long term, 10 years and more after gastric resection (39 men).
Previous papers dealt with the discovery that hydrolysis of 17-hydroxycorticosteroid glucuronides in urine with beta-glucuronidase preparations from bovine liver is increased by adding sodium sulfate to the incubation medium. Here, we conclude that sodium sulfate not only increases the activity of bovine liver beta-glucuronidase on 17-hydroxycorticosteroid glucuronides and p-nitrophenyl glucuronides, but also removes the inhibitory activity of substances of high molecular weight in urine and, moreover, inhibits urine putrefaction during the hydrolysis. The net effect is an increased yield of urinary 17-hydroxycorticosteroids. In the incubation with sodium sulfate (Na2SO4, final concentration 80 g/liter, 500 Fishman units of beta-glucuronidase per milliliter of urine, pH 5.0, 48 degrees C, 18 h) the analytical recovery of 17-hydroxycorticosteroid glucuronides added to 12 urine samples proved to be 98 +/- 1.8% (95-100).
We evaluated the usefulness of the basal urinary 24-h excretion rates of free cortisol versus 17-hydroxycorticosteroids in the diagnosis of Cushing's syndrome. On an outpatient basis, both urinary free cortisol and 17-hydroxycorticosteroids levels were determined in 48 patients with Cushing's syndrome, as well as in 95 obese and 94 healthy control persons of normal weight. Determination of the urinary free cortisol content allowed a clear-cut distinction between the patients with hypercortisolism and the controls, resulting in a sensitivity of 100% and specificity of 98% for the diagnosis of Cushing's syndrome. The diagnostic accuracy of urinary free cortisol was distinctly superior to that of 17-hydroxycorticosteroids, which showed a wide overlap of values between the groups, with a sensitivity of 73% and a specificity of 94%. In conclusion, the measurement of basal urinary free cortisol provided an excellent diagnostic sensitivity and specificity in the assessment of adrenocortical function. This simple and accurate test thus seems to be particularly useful in the outpatient evaluation of patients with suspected Cushing's syndrome.
BACKGROUND: Although many workers have tested adrenal function in the elderly, few have studied the effect of aging on cortisol production rate or urinary free cortisol or 6 beta-hydroxycortisol excretion, and none have published comparisons of these variables between old people of defined health status and young people. METHODS: We have measured cortisol production rate and the urinary excretion of free cortisol, 6 beta-hydroxycortisol, 17-hydroxycorticosteroids (Porter-Silber chromogens) and creatinine in elderly men and women screened by the SENIEUR protocol and in young men; 17-hydroxycorticosteroid and 6 beta-hydroxycortisol excretion were also measured in young women. The period of measurement was 24 h or, usually, 48 h. RESULTS: Only 6 beta-hydroxycortisol excretion was affected by aging; it was lower in the elderly men and women than in their younger counterparts. Urinary free cortisol excretion was lower in the elderly women than in the elderly men. There were no significant differences between groups in cortisol production rate or 17-hydroxycorticosteroid excretion. Excretion and (over the first 24 h) clearance of creatinine were lower in the old women than in the old men. The cortisol-related variables tended to be positively correlated with each other and with the relevant creatinine-related variables in the elderly subjects; over the first but not the second 24 h, most of the correlations were significant in the men and women combined. CONCLUSIONS: With the exception of 6 beta-hydroxycortisol, the data agree with measurements of plasma cortisol and the results of adrenal function tests in showing little change in hypothalamic-pituitary-adrenal function with aging in healthy people.
Urinary levels of free adrenaline, noradrenaline and 11-hydroxycorticosteroid were measured in 30 surgeons while they operated, to test the hypothesis that sympathetic and adrenocortical systems are overactivated by occupational environment of surgery procedures. They were divided in two groups according to a crossover design in which the surgeons had taken alternatively an oral dose of 40 mg oxprenolol and placebo. During operations with placebo urinary free adrenaline, noradrenaline and 11-hydroxycorticosteroid rose significantly compared with the control values obtained during ward-round performed by the same surgeons. The increase of adrenaline, noradrenaline and 11-hydroxycorticosteroid was abolished by 40 mg of oxprenolol which contained their urinary concentration to the control values. These data support the concept that operating stress of surgeons influences the adrenosympathetic and adrenocortical systems and they indicate a possible role of beta-blocking drugs in minimizing the adverse reactions of a such hormonal overactivity.