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1, 2, 3, 4-13C cortisol(13C-F) was used to measure the CPR in two sisters(karyotype 46, XX)of 11(I) and 3(II) yrs of age with 17α-hydroxylase deficiency. The disease was diagnosed on hyper tension, low plasma cone, of 17β-estradiol, cortisol, PRA and aldosterone and high conc, of progesterone and in patient I of ACTH, FSH and LH. Both children excreted in the urine high amounts of tetra-and hexahydro-metabolites of corticosterone and deoxycorticosterone and (very) low amounts of metabolites of cortisol (F), androstenedione and testosterone. After the i.v. dosage of 60 and 2.4 μg to patient I and II, urine was collected for 3 days. After extraction, hydrolysis and HPLC Chromatography the metabolites of F were oxidised to 11-oxoetiocholanolone (11-OET) and -androsterone. The 13C-enrichments in the methoxime-tert.butyldimethylsily ethers were measured by gas Chromatography mass fragmentography at the leading ions m/z 348 (13C4) and 344 (12C). Calibration standards were prepared from mixtures of 13C-F and F, which were oxidised to cortisone (E), reduced to tetrahydrocortisone (THE) by Cl.paraputrificum and oxidised to 11-OET. The CPR (mg/day) was for I:0.28±0.003 (THE) and 0.19±0.007 (α+βcortolone) and for II:0.022±0.003 (THE). Owing to the very low amounts of cortisol metabolites the CPR determinations would not have been possible if tritiated cortisol was used. Financial support was obtained from the Netherlands Organisation for the Advancement of Pure Research (ZWO).

Recently we have published detailed methodology for the comprehensive analysis by capillary gas chromatography of all the major urinary androgen, progestin and corticoid metabolites (G. Phillipou et al., Aust. N.Z. J. Med., 1978, 8, 63). Between assay variability (mean, μmol/24h; C.V.) for several steroids as determined by this procedure in a control urine (N = 8; 10 ml, TV = 2260 ml) was: androsterone, 11.0, 12.5%; pregnanetriol, 2.4 12.5%; tetrahydrocortisone, 10.4, 10%. Patient samples (female 463, male 31) referred for analysis may be classified into 2 groups. Group A (9%) comprises disorders such as inborn errors of corticoid biosynthesis and adrenal hyperfunction which are uniquely characterized by the selective and marked elevation of certain urinary steroids. Group B (91%) encompasses more common endocrine disorders represented by amenorrhoea, hirsutism, obesity and alopecia which cannot be defined by any one urinary steroid. Consequently, evaluation of these latter patients is complicated and contingent upon reference to a normal range.

Urinary steroids of 12 children were assayed as their MO-TMSi-derivatives by gas-liquid chromatography on open tubular columns coated with SE 30 as stationary phase. Extracts were separated on SEPHADEX LH 20, before cleavage of conjugates, into free steroids, glucosiduronates, and sulfates.-The output of 11-desoxy-, and particularly ll-oxygenated-17-oxo-steroids was markedly reduced, most significantly in the glue.-fraction; the decrease in 17-hydroxycorticosteroids was mainly due to low tetrahydrocortisone and cortolones; the percentage contribution of 5-alpha-tetra-hydrocortisol was decreased to 40% of normal. Under ACTH-stimulation the 5-alpha/5-beta ratio for androsterone/etiocholanolone and 5-alpha-tetrahydrocortisol/tetrahydrocortisol was further lowered (.3) than without (.5) or later during thyroid replacement therapy (1.1;.9). The percentage distribution of 17-alpha-hydroxycorticosteroide in fractions gluc-sulf.-free was 70/20/10%, under ACTH 60/20/20%.- These results comply with recent observations in adults indicating a diminished activity of 11-beta-hydroxy-steroid dehydrogenase, 5-alpha-reductase, and glucuronyl-transferase-system in hypothyroidism. Supported by Deutsche Forschungsgemeinschaft Bl 95/4.

In a previous investigation1analysis of chloroform extracts of aqueous humor from normal rabbit eyes by the blue tetrazolium method indicated the presence of corticosteroids in quantities of the order of magnitude of 1-1.5γ per milliliter. Since the blue tetrazolium method neither permits the identification of individual compounds in a mixture nor is specific only for oxidizable corticoids, the extracted aqueous humor was fractionated by paper partition chromatography and analyzed for corticosteroids. The results of this investigation strongly indicate that at least part of the blue-tetrazoliumpositive material present in aqueous humor is hydrocortisone. MATERIALS The following purified crystalline corticosteroids were donated by Sharp & Dohme, Division of Merck & Company, Inc.: 11-dehydrocorticosterone (A); corticosterone (B); 11-desoxy-17-hydroxycorticosterone (S); cortisone (11-dehydro-17hydroxycorticosterone, Compound E); hydrocortisone (17-hydroxycorticosterone, Compound F); hydrocortisone acetate; dihydrocortisone, and tetrahydrocortisone. Small aliquots of the samples were dissolved in absolute methanol in concentrations of 1γ and 37 per microliter and

We have studied the urinary excretion of cortisol (F), cortisone (E), tetrahydrocortisol (THF), tetrahydrocortisone (THE), 6β-hydroxycortisol (6β-OHF) and corticosterone (B) in premature infants. Each specimen was extracted sequentially for free, β-glucuronidase and solvolysis liberated steroids; each metabolite was identified by constant isotope ratios, and quantitated by a double-isotope derivative assay. In 3 normal prematures studied within the first 48 hours of life, 6β-OHF, THF, F and B were mainly recovered as free or sulfated compounds. THE and E were mainly sulfurylated. In a 4th infant, with respiratory distress syndrome, studied similarly, 6β-OHF was mainly sulfated while THF was mostly as a glucuronoside. THE and B were equally glucuro and sulfo-conjugated while F and E were equally recovered in free or sulfated form. In a pool of 8 premature infants studied from two weeks of age, prior to ACTH, approximately 60 % of F and E were sulfo-conjugated, while a larger proportion of B was in the sulfate form. 6β-OHF and THF were predominantly excreted as sulfates, while THE was mostly unconjugated. Post ACTH, there was a considerable increase of all steroids and glucuro-conjugation seemed genererally enhanced except for F. These studies suggest that: 1. in resting state, most C21 steroids are excreted unconjugated or conjugated with sulfuric acid, in accord with limited glucuro-conjugation; 2. some limitation in reduction of Ring A of the steroid molecule exists since more unreduced metabolites are recovered than in older children and adults; 3. glucuroconjugation can be enhanced under stress or ACTH, suggesting that glucuro-conjugation is not maximal in resting state; 4. these metabolic particularities persist at least up to four weeks of age. (SPR)