Download PDF
Horm Metab Res 2003; 35(7): 421-426
DOI: 10.1055/s-2003-41623
Original Clinical
© Georg Thieme Verlag Stuttgart · New York

Plasma and Salivary 6β-Hydroxycortisol Measurements for Assessing Adrenocortical Activity in Patients with Adrenocortical Adenomas

I.  Varga1 , C.  Jakab1 , N.  Szücs1 , A.  Patócs1 , M.  Tóth1 , R.  Kiss1 , E.  Gláz1 , K.  Rácz1
  • 1Gastroenterological and Endocrinological Research Group, 2nd Department of Medicine, Faculty of Medicine, Semmelweis University, Budapest, Hungary
Further Information

Publication History

Received 23 October 2002

Accepted after Revision 27 March 2003

Publication Date:
21 August 2003 (online)

Also available at
    Permissions and Reprints

Abstract

The aim of this study was to examine and compare the potential usefulness of plasma and salivary 6β-hydroxycortisol measurements for assessing adrenocortical activity in patients with adrenocortical adenomas. Plasma and salivary cortisol as well as 6β-hydroxycortisol determinations were performed by radioimmunoassay after extraction with ethyl acetate followed by chromatographic separation using a modified paper chromatographic system. Samples were obtained from 36 control subjects and 37 patients with non-hyperfunctioning adrenocortical adenomas in the morning at 8 a. m. after a low-dose of dexamethasone and after stimulation with synthetic depot ACTH. Basal and post-dexamethasone hormone levels were also measured in plasma and salivary samples of 4 patients with Cushing's syndrome from adrenal adenomas. In the baseline state, patients with non-hyperfunctioning adrenocortical adenomas had significantly higher plasma and salivary 6β-hydroxycortisol levels (mean ± SE, 79.0 ± 7 and 17.1 ± 2.2 ng/dl, respectively) compared to those measured in controls (62.0 ± 4 and 7.7 ± 0.6 ng/dl, respectively), whereas baseline plasma and salivary cortisol levels (9.6 ± 0.5 µg/dl and 342 ± 39 ng/dl, respectively) were similar to those measured in the control group (9.9 ± 0.4 µg/dl and 366 ± 24 ng/dl, respectively). In all groups, the changes in plasma and salivary 6β-hydroxycortisol concentrations after dexamethasone suppression and ACTH stimulation were similar to the changes in plasma and salivary cortisol levels, although the differing ratios of 6βOHF to cortisol indicated potentially important variations in the induction of 6β-hydroxylase activity between the three groups. In patients with Cushing's syndrome, baseline plasma and salivary 6β-hydroxycortisol concentrations (754 ± 444 and 104 ± 88 ng/dl, respectively) were more markedly increased than plasma and salivary cortisol levels (24.8 ± 6.7 µg/dl and 1100 ± 184 ng/dl, respectively), and all remained non-suppressible after dexamethasone administration. These results suggests that plasma and salivary 6β-hydroxycortisol determinations may precisely detect not only overt increases of cortisol secretion in patients with Cushing's syndrome but also mild glucocorticoid overproduction presumably present in patients with non-hyperfunctioning adrenocortical tumors.

Key words

Cushing's syndrome - Non-hyperfunctioning adrenocortical adenoma - Cortisol metabolism - ACTH - Dexamethasone

References

  • 1 Vining R F, McGinley R A, Maksvytis J J, Ho K Y. Salivary cortisol: a better measure of adrenal cortical functions than serum cortisol.  Ann Clin Biochem. 1983;  20 329-335
    PubMedGoogle Scholar
  • 2 Aardal-Eriksson E, Karlberg B E, Holm A-C. Salivary cortisol - an alternative to serum cortisol determination in dynamic function tests.  Clin Chem Lab Med. 1998;  36 215-222
    CrossrefPubMedGoogle Scholar
  • 3 Barrou Z, Guiban D, Maroufi A, Fournier C, Dugué M A, Luton J P, Thomopoulos P. Overnight dexamethasone suppression test: comparison of plasma and salivary cortisol measurement for the screening of Cushing's syndrome.  Eur J Endocrinol. 1996;  134 93-96
    CrossrefPubMedGoogle Scholar
  • 4 Laudat M H, Cerdas S, Fournier C, Guiban B, Guilhaume B, Luton J P. Salivary cortisol measurement: a practical approach to assess pituitary-adrenal function.  J Clin Endocrinol. Metab1988;  66 343-348
    CrossrefPubMedGoogle Scholar
  • 5 Francis S J, Walker R F, Riad-Fahmy D, Hughes D, Murphy J F, Gray O P. Assessment of adrenocortical activity in term newborn infants using salivary cortisol determinations.  J Pediatrics. 1987;  111 129-133
    PubMedGoogle Scholar
  • 6 Antonini S RR, Jorge S M, Moreira A C. The emergence of salivary cortisol circadian rhythm and its relationship to sleep activity in preterm infants.  Clin Endocrinol. 2000;  52 423-426
    CrossrefPubMedGoogle Scholar
  • 7 Martinelli C E, Sader S L, Oliveira E B, Daneluzzi J C, Moreira A C. Salivary cortisol for screening of Cushing's syndrome in children.  Clin Endocrinol. 1999;  51 67-71
    CrossrefPubMedGoogle Scholar
  • 8 Nelson N, Arbring K, Theodorsson E. Neonatal salivary cortisol in response to heelstick: method modifications enable analysis of low concentrations and small sample volumes.  Scand J Clin Lab Invest. 2001;  61 287-291
    PubMedGoogle Scholar
  • 9 Luthold W W, Marcondes J AM, Wajchenberg B L. Salivary cortisol for the evaluation of Cushing's syndrome.  Clin Chim Acta. 1985;  151 33-39
    CrossrefPubMedGoogle Scholar
  • 10 Raff H, Raff J L, Findling J W. Late-night salivary cortisol as a screening test for Cushing's syndrome.  J Clin Endocrinol Metab. 1998;  83 2681-2686
    CrossrefPubMedGoogle Scholar
  • 11 Strickland P, Morris A, Wearden A, Deakin B. A comparison of salivary cortisol in chronic fatigue syndrome, community depression and healthy controls.  J Affect Dis. 1998;  47 191-194
    PubMedGoogle Scholar
  • 12 Castro M, Elias P CL, Quidute A RP, Halah F PB, Moreira A C. Out-patient screening for Cushing's syndrome: the sensitivity of the combination of circadian rhythm and overnight dexamethasone suppression salivary cortisol tests.  J Clin Endocrinol Metab. 1999;  84 878-882
    CrossrefPubMedGoogle Scholar
  • 13 Immanuel C, Sarma G R, Krishnamurthy P V, Ramachandran G, Kumaraswami V. Salivary cortisol in the assessment of adrenocortical functions in patients with pulmonary tuberculosis.  Indian J Med Res (A). 1992;  95 1-7
    PubMedGoogle Scholar
  • 14 Hucklebridge F, Sen S, Evans P D, Clow A. The relationship between circadian patterns of salivary cortisol and endogenous inhibitor monoamine oxidase A.  Life Sci. 1998;  62 2321-2328
    CrossrefPubMedGoogle Scholar
  • 15 Lipman M M, Katz F H, Jailer J W. An alternative pathway for cortisol metabolism: 6β-hydroxycortisol production by human tissue slices.  J Clin Endocrinol Metab. 1962;  22 268-272
    PubMedGoogle Scholar
  • 16 Burstein S, Dorfman R I, Nadel E M. 6-beta-hydroxycortisol - a new steroid in human urine.  Arch Biochem and Biophys. 1954;  53 307-312
    CrossrefPubMedGoogle Scholar
  • 17 Katz F H, Lipman M M, Frantz A G, Jailer J W. The physiologic significance of 6β-hydroxycortisol in human corticoid metabolism.  J Clin Endocrinol Metab. 1962;  22 71-77
    PubMedGoogle Scholar
  • 18 Bienvenu T, Rey E, Pons G, Athis P D, Olive G. A simple non-invasive procedure for the investigation pf cytochrome P-450 IIIA dependent enzymes in human.  Int J Clin Pharmacol Ther Toxicol. 1991;  29 441-445
    PubMedGoogle Scholar
  • 19 Hunter D JS, Keane P, Walker W HC, YoungLai E V. Variations in urinary levels of free 6β-hydroxycortisol, cortisol and estrogens in late pregnancy.  Gynecol Obstet Invest. 1984;  18 83-87
    PubMedGoogle Scholar
  • 20 Patel S B, Toddywalla V S, Betrabet S S, Kulharni R D, Kombo I, Saxena B N. Age related changes in urinary 6-beta hydroxycortisol in normal infants.  Indian Pediatr. 1996;  33 398-401
    PubMedGoogle Scholar
  • 21 Voccia E, Saenger P, Peterson R E, Rauh W, Gottesdiener K, Levine L S, New M I. 6β-hydroxycortisol excretion in hypercortisolemic states.  J Clin Endocrinol Metab. 1979;  48 467-471
    PubMedGoogle Scholar
  • 22 Edwards O M, Courtenay-Evans R J, Galles J M, Hunter J, Tait A D. Changes in cortisol metabolism following rifampicin therapy.  Lancet. 1974;  2 549-551
    PubMedGoogle Scholar
  • 23 Saenger P, Forster E, Kream J. 6β-hydroxycortisol: a non-invasive indicator of enzyme induction.  J Clin Endocrinol Metab. 1981;  52 381-384
    PubMedGoogle Scholar
  • 24 Moreland T A, Park B K, Rylance G W. Microsomal enzyme induction in children: the influence of carbamazepine treatment on antipyrine kinetics, 6-beta-hydroxycortisol excretion and plasma gamma-glutamyltranspeptidase activity.  Br J Clin Pharmacol. 1982;  14 861-865
    PubMedGoogle Scholar
  • 25 Burstein S, Kimball H I, Klaiber E L, Gut M. Metabolism of 2-alpha- and 6β-hydroxycortisol in man: determination of production rates of 6β-hydroxycortisol with and without phenobarbital administration.  J Clin Endocrinol Metab. 1967;  27 491-499
    PubMedGoogle Scholar
  • 26 Fukushima D K, Bradlow H L, Hellman L. Effects of o,p'-DDD on cortisol and 6β-hydroxycortisol secretion and metabolism in man.  J Clin Endocr. 1971;  32 192-200
    PubMedGoogle Scholar
  • 27 Nahoul K, Adeline J, Paysant J, Scholler R. Radioimmunoassay of plasma and urine 6β-hydroxycortisol: levels in healthy adults and in hypercortisolemic states.  J Steroid Biochem. 1982;  17 343-350
    CrossrefPubMedGoogle Scholar
  • 28 Nakamura J, Minoru Y. Assessing adrenocortical activity by determining levels of urinary free cortisol and urinary 6β-hydroxycortisol.  Acta Endocrinol (Copenh). 1989;  120 277-283
    PubMedGoogle Scholar
  • 29 Kishida S, Fukushima D K. Radioimmunoassay of 6β-hydroxycortisol in human plasma and urine. Steroids.  1977;  30 741-749
    PubMedGoogle Scholar
  • 30 Vecsei P, Onyechi J, Hornung J, Dietz R, Mast G, Hobler H. Use of corticosteroid antibodies for the study of corticosteroid biosynthesis in vitro.  J Steroid Biochem. 1975;  6 383-387
    CrossrefPubMedGoogle Scholar
  • 31 Conolly T M, Vecsei P. Simple radioimmunoassay of cortisol in diluted samples of human plasma.  Clin Chem. 1978;  24 1466-1472
    PubMedGoogle Scholar
  • 32 Park B K. Assessment of urinary 6β-hydroxycortisol as an in vivo index of mixed function oxygenase activity.  Br J Clin Pharmac. 1981;  12 97-102
    PubMedGoogle Scholar
  • 33 Touchstone J C, Kasparow M, Rosenthal O. Most polar steroids of human adrenal incubates.  Fed Proc. 1959;  18 340
    PubMedGoogle Scholar
  • 34 Gláz E, Rácz K, Varga I, Kiss R, Tóth M, Fütö L. Mineralocorticoid production of adrenal cortical adenomas.  J Steroid Biochem. 1993;  45 57-64
    PubMedGoogle Scholar
  • 35 Ambrosi B, Peverelli S, Passini E. Re T, Ferrario R, Colombo P. Abnormalities of endocrine function in patients with clinically ‘silent' adrenal masses.  Eur J Endocrinol. 1995;  132 22-428
    PubMedGoogle Scholar
  • 36 Tóth M, Rácz K, Varga I, Adleff V, Jakab C, Fütö L, Kiss R, Gláz E. Plasma dehydroepiandrosterone sulfate levels in patients with hyperfunctioning and non-hyperfunctioning adrenal tumors before and after adrenal surgery.  Eur J Endocrinol. 1997;  136 290-295
    PubMedGoogle Scholar
  • 37 Reincke M, Nieke J, Krestin G P, Saeger W, Allolio B, Winkelmann W. Preclinical Cushing's syndrome in adrenal ”incidentalomas”: comparison with adrenal Cushing's syndrome.  J Clin Endocrinol Metab. 1992;  75 826-832
    CrossrefPubMedGoogle Scholar
  • 38 Ross N S. Epidemiology of Cushing's syndrome and subclinical disease.  Endocrinol Metab Clin North Am. 1994;  23 539-546
    PubMedGoogle Scholar
  • 39 Tersolo M, Osella G, Ali A, Borretta G, Cesario F, Paccotti P, Angeli A. Subclinical Cushing's syndrome in adrenal incidentalomas.  Clin Endocrinol (Oxf). 1998;  48 89-97
    Google Scholar
  • 40 Zhiri A, Mayer H A, Michaux V, Wellman-Bednawska M, Slest G. 6β-hydroxycortisol in serum and urine determined by enzyme immunoassay on microtitre plates.  Clin Chem. 1986;  30 2094-2097
    PubMedGoogle Scholar
  • 41 Park B K. A direct radioimmunoassay for 6β-hydroxycortisol in human urine.  J Steroid Biochem,. 1978;  9 963-966
    CrossrefPubMedGoogle Scholar
  • 42 Ng M CY, Young R P, Critchley J AJH, Leung N WY, Lau J WY, Li A KC. Urinary 6β-hydroxycortisol excretion in Hong Kong Chinese patients with hepatocellular carcinoma and other chronic liver diseases.  Cancer,. 1996;  77 1427-1433
    CrossrefPubMedGoogle Scholar
  • 43 Tang C, Kassahun K, McIntosh I S, Brunner J, Rodrigues A D. Simultaneous determination of urinary free cortisol and 6β-hydroxycortisol by liquid chromatography - atmospheric pressure chemical ionization tandem mass spectrometry and its application for estimating hepatic CYP3A induction.  J Chromatogr B. 2000;  742 303-313
    PubMedGoogle Scholar
  • 44 Furukori Y N, Kondo T, Kubota T, Otake H, Ohkubo T, Nagasaki T, Sugawara K, Chiba K, Otani K, Kaneko S. No correlations between the urinary ratio of 6 beta-hydroxycortisol to free cortisol and pharmacokinetics of alprazolam.  Eur J Clin Pharmacol. 2001;  57 285-288
    CrossrefPubMedGoogle Scholar

Dr. K. Rácz

2nd Department of Medicine · Faculty of Medicine · Semmelweis University

Szentkirályi 46 · Budapest · 1088 Hungary

Phone: + 361 (266) 0926

Fax: + 361 (266) 0816

Email: racz@bel2.sote.hu

      >