Simultaneous determination of cortisol and cortisone in urine by reversed-phase high-performance liquid chromatography clinical and doping control applications

Santos-Montes, A.; Gonzalo-Lumbreras, R.; Izquierdo-Hornillos, R.

doi:10.1016/0378-4347(95)00253-f

Cited by 31 publications

(18 citation statements)

References 23 publications

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“…Twenty-four hours UFC was assessed by HPLC in accordance with the modified Santos-Montes method (intra-assay CV 7.73%; normal range 9.2-45.2 mg/24 h) (33).…”

Section: Assaysmentioning

confidence: 99%

Association of glucocorticoid receptor polymorphism A3669G with decreased risk of developing diabetes in patients with Cushing's syndrome

Trementino

Appolloni

Concettoni

et al. 2012

European Journal of Endocrinology

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Objective: Glucocorticoid receptor (GR) polymorphisms alter glucocorticoid (GC) sensitivity and have been associated with altered metabolic profiles. We evaluate the prevalence of the four GR (NR3C1) polymorphisms BclI, N363S, ER22/23EK, and A3669G in patients with Cushing's syndrome (CS) compared with healthy controls (HC) and we investigate their role in the development of metabolic abnormalities in patients with CS according to their hormonal profile. Patients and methods: Sixty-one patients with CS and 71 sex-and age-matched HC were genotyped. Results: BclI variant was markedly higher in patients with CS compared with HC (62 vs 41%, P!0.05) while no significant differences were found among other polymorphisms. A very low frequency of N363S and the ER22/23EK was observed. In CS patients, despite the significantly increased levels of morning serum cortisol in BclI carriers compared with wild type no clinical or metabolic differences were found. In contrast, A3669G GR carriers showed a significantly reduced prevalence of type 2 diabetes mellitus compared with wild type (19 vs 68%, PZ0.001) despite the higher levels of both serum morning (21.7G6 vs 27.3G8.6 mg/dl, PZ0.009) and midnight cortisol (18.8G5.8 vs 24.0G8.0 mg/dl, PZ0.01). The negative association between diabetes and A3669G GR polymorphism remained significant when data were adjusted for potential confounding factors. Conclusions: The A3669G polymorphism of the GR gene plays a protective role in patients with CS, attenuating the effects of GC excess on glucose metabolism as shown by their reduced risk of diabetes.

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“…Twenty-four hours UFC was assessed by HPLC in accordance with the modified Santos-Montes method (intra-assay CV 7.73%; normal range 9.2-45.2 mg/24 h) (33).…”

Section: Assaysmentioning

confidence: 99%

Association of glucocorticoid receptor polymorphism A3669G with decreased risk of developing diabetes in patients with Cushing's syndrome

Trementino

Appolloni

Concettoni

et al. 2012

European Journal of Endocrinology

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“…33 Briefly, 200 ml serum or urine were mixed with 200 ml 0.2 M sodium hydrogen carbonate (pH 9.6) and 25 ml methylprednisolone (50 ng, internal standard solution), and extracted with 2 ml dichloromethane. The organic layer was evaporated to dryness and the residue was reconstituted with 100 ml mobile phase, of which 50 ml was injected into the HPLC system.…”

Section: Laboratory Parametersmentioning

confidence: 99%

Renal clearance and daily excretion of cortisol and adrenal androgens in patients with rheumatoid arthritis and systemic lupus erythematosus

Straub¹

2004

Annals of the Rheumatic Diseases

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Background: In rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE), patients demonstrate low levels of adrenal hormones. Objective: To investigate whether increased renal clearance and daily excretion contribute to this phenomenon. Methods: Thirty patients with RA, 32 with SLE, and 54 healthy subjects (HS) participated. Serum and urinary levels of cortisol, cortisone, 17-hydroxyprogesterone (17OHP), androstenedione, dehydroepiandrosterone (DHEA), and DHEA sulphate (DHEAS) were measured. Results: Clearance of DHEAS and DHEA was lower in patients than in HS, and clearance of androstenedione was somewhat higher in patients than in HS, but daily excretion of this latter hormone was low. Clearance of cortisol, cortisone, and 17OHP was similar between the groups. The total molar amount per hour of excreted DHEA, DHEAS, and androstenedione was lower in patients than HS (but similar for cortisol). Serum DHEAS levels correlated with urinary DHEAS levels in HS and patients, whereby HS excreted 5-10 times more of this hormone than excreted by patients. Low serum levels of adrenal androgens and cortisol in patients as compared with HS were confirmed, and proteinuria was not associated with changes of measured renal parameters. Conclusions: This study in patients with RA and SLE demonstrates that low serum levels of adrenal androgens and cortisol are not due to increased renal clearance and daily loss of these hormones. Decreased adrenal production or increased conversion or conjugation to downstream hormones are the most likely causes of inadequately low serum levels of adrenal hormones in RA and SLE.

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“…The use of HPLC has been reported, but we have found that a number of methods were not suitable for the urine samples under investigation (Gatti et al, 2005;Hay and Mormede, 1997;Izquierdo-Hornillis et al, 2005;Santos-Montes et al, 1995;Vogeser and Jacob, 2000;Yao and Zhang, 2001). Certainly, any method involving the use of a single HPLC column for separation proved unsuccessful in our hands.…”

Section: Introductionmentioning

confidence: 99%

An HPLC method for the determination of the free cortisol/cortisone ratio in human urine

Sharef

Feely

Kavanagh

et al. 2007

Biomedical Chromatography

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The measurement of the urinary free cortisol-cortisone ratio has been reported to be a sensitive indicator of renal 11 beta-hydroxysteroid dehydrogenase type 2 (11 beta-HSD 2) activity. This converts biologically active cortisol to inactive cortisone. A decrease in its activity (e.g. through disease or inhibition caused by a therapeutic agent or a foodstuff) may increase cortisol levels and susceptibility towards hypertension. The method presented here uses a simple isocratic tandem column HPLC system. The method has been validated and found to be robust and reproducible. The lower limit of quantification (LLOQ) was found to be 10 ng/mL for both cortisol and cortisone. Samples of urine (n = 99) from patients, most of whom were on complex combinations of drugs, were analyzed and 92% of samples were found to give successful results with this method (cortisol and cortisone above LLOQ). The ratio ranged from 0.07 to 5.61. No interferences were noted from the drugs that the patients were taking. It was also found that a morning spot urine sample gave comparable results to 24 h collection samples, thus making sample collection much easier.

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Simultaneous determination of cortisol and cortisone in urine by reversed-phase high-performance liquid chromatography clinical and doping control applications

Cited by 31 publications

References 23 publications

Association of glucocorticoid receptor polymorphism A3669G with decreased risk of developing diabetes in patients with Cushing's syndrome

Association of glucocorticoid receptor polymorphism A3669G with decreased risk of developing diabetes in patients with Cushing's syndrome

Renal clearance and daily excretion of cortisol and adrenal androgens in patients with rheumatoid arthritis and systemic lupus erythematosus

An HPLC method for the determination of the free cortisol/cortisone ratio in human urine

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