Paediatric reference range for overnight urinary cortisol corrected for creatinine

Abstract Objectives Systemic activity of inhaled corticosteroids (ICS) may be assessed via urinary cortisol measurement. Overnight urinary free cortisol corrected for creatinine (OUFCC) has been extensively reported in adult studies. However, a paediatric mass spectrometric (MS) reference range for OUFCC is not established. MS methods for OUFCC avoid cross-reactivity with other steroid hormones and are thus preferable to immunoassays. The aim of the present study was to define an MS OUFCC normative range in children. Methods This was a cross-sectional study of healthy pre-pubertal children from 5 to 11 years. Children collected urine from 10 pm or bedtime, whichever was earlier, until 8 am. Urinary free cortisol was measured via a liquid chromatography tandem mass spectrometry (LC-MS/MS) assay (Acquity UPLC with Xevo TQ-S Mass Spectrometer [Waters]) with in-house reagents. Urinary creatinine was measured using a commercial assay (Roche). Results Complete urine collections were obtained from 72 males and 70 females, mean age (SD) 8.6 (1.9) (range 5.0–11.8) years. The OUFCC 95% prediction interval was 1.7–19.8 nmol/mmol. Geometric mean OUFCC was 5.7; range 1.1–24.8 nmol/mmol. Conclusions The obtained normative LC-MS/MS OUFCC reference data facilitate the use of mass spectrometry OUFCC assays in assessment of systemic activity of endogenous and exogenous corticosteroids in children.

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Urinary sodium is positively associated with urinary free cortisol and total cortisol metabolites in a cross-sectional sample of Australian schoolchildren aged 5–12 years and their mothers

British Journal Of Nutrition ◽

10.1017/s0007114518003148 ◽

2018 ◽

Vol 121 (2) ◽

pp. 164-171

Author(s):

Susan J. Torres ◽

Carley Grimes ◽

Caryl A. Nowson ◽

Sisitha U. Jayasinghe ◽

Clinton R. Bruce ◽

...

Keyword(s):

Primary Schools ◽

Regression Models ◽

Urinary Free Cortisol ◽

Linear Regression Models ◽

Urinary Cortisol ◽

Cross Sectional ◽

Free Cortisol ◽

Potential Impact ◽

Cortisol Levels ◽

Cortisol Metabolites

AbstractHigh Na intake and chronically elevated cortisol levels are independently associated with the development of chronic diseases. In adults, high Na intake is associated with high levels of urinary cortisol. We aimed to determine the association between urinary Na and K and urinary cortisol in a cross-sectional sample of Australian schoolchildren and their mothers. Participants were a sample of Australian children (n 120) and their mothers (n 100) recruited through primary schools. We assessed Na, K, free cortisol and cortisol metabolites in one 24 h urine collection. Associations between 24 h urinary electrolytes and 24 h urinary cortisol were assessed using multilevel mixed-effects linear regression models. In children, urinary Na was positively associated with urinary free cortisol (β=0·31, 95 % CI 0·19, 0·44) and urinary cortisol metabolites (β=0·006, 95 % CI 0·002, 0·010). Positive associations were also observed between urinary K and urinary free cortisol (β=0·65, 95 % CI 0·23, 1·07) and urinary cortisol metabolites (β=0·02, 95 % CI 0·03, 0·031). In mothers, urinary Na was positively associated with urinary free cortisol (β=0·23, 95 % CI 0·01, 0·50) and urinary cortisol metabolites (β=0·008, 95 % CI 0·0007, 0·016). Our findings show that daily Na and K intake were positively associated with cortisol production in children and their mothers. Investigation of the mechanisms involved and the potential impact of Na reduction on cortisol levels in these populations is warranted.

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Pseudo-Cushing Syndrome Caused by Fenofibrate Interference with Urinary Cortisol Assayed by High-Performance Liquid Chromatography

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/jc.2003-030234 ◽

2003 ◽

Vol 88 (8) ◽

pp. 3521-3524 ◽

Cited By ~ 47

Author(s):

A. Wayne Meikle ◽

James Findling ◽

Mark M. Kushnir ◽

Alan L. Rockwood ◽

Gordon J. Nelson ◽

...

Keyword(s):

Mass Spectrometry ◽

High Performance ◽

Cushing Syndrome ◽

Urinary Free Cortisol ◽

Urinary Cortisol ◽

Free Cortisol ◽

Hplc Peak ◽

Major Transition ◽

Single Transition ◽

Mass Spectrometry Mass Spectrometry

Urinary free cortisol (UFC) excretion over 24 h reflects the production rate of cortisol and is used commonly in the diagnosis of Cushing syndrome. We report on two patients evaluated for Cushing syndrome who had elevated UFC when analyzed by HPLC but normal values for the analysis performed by RIA and HPLC-mass spectrometry/mass spectrometry (HPLC-MS/MS). Other laboratory testing was inconsistent with the diagnosis of Cushing syndrome and raised doubts about the diagnosis. We identified a probable cause of analytical interference as coming from fenofibrate (Tricor), medication taken by the patients. Fenofibrate peak overlapped with the HPLC peak of cortisol and produced an MS/MS transition overlapping the major transition of cortisol. A second MS/MS transition was free from interference. In summary, fenofibrate administration may cause false elevation of UFC values determined by HPLC or HPLC-MS/MS in patients evaluated for Cushing syndrome. An HPLC-MS/MS method using multiple mass transitions, rather than a single transition, allows accurate quantitation of urinary cortisol in patients taking fenofibrate.

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Validation of a High-Throughput Liquid Chromatography–Tandem Mass Spectrometry Method for Urinary Cortisol and Cortisone

Clinical Chemistry ◽

10.1093/clinchem/48.9.1511 ◽

2002 ◽

Vol 48 (9) ◽

pp. 1511-1519 ◽

Cited By ~ 148

Author(s):

Robert L Taylor ◽

Dwaine Machacek ◽

Ravinder J Singh

Keyword(s):

Mass Spectrometry ◽

Liquid Chromatography ◽

Tandem Mass Spectrometry ◽

High Throughput ◽

Urinary Free Cortisol ◽

Tandem Mass ◽

Urinary Cortisol ◽

Chromatography Tandem Mass Spectrometry ◽

Free Cortisol ◽

Liquid Chromatography Tandem Mass

Abstract Background: Urinary free cortisol and cortisone measurements are useful in evaluation of Cushing syndrome, apparent mineralocorticoid excess, congenital adrenal hyperplasia, and adrenal insufficiency. To reduce analytical interference, improve accuracy, and shorten the analysis time, we developed a liquid chromatography–tandem mass spectrometry (LC-MS/MS) method for urinary cortisol and cortisone. Methods: We added 190 pmol (70 ng) of stable isotope cortisol-9,11,12,12-d4 to 0.5 mL of urine as an internal standard before extraction. The urine was extracted with 4.5 mL of methylene chloride, washed, and dried, and 10 μL of the reconstituted extract was injected onto a reversed-phase C18 column and analyzed using a tandem mass spectrometer operating in the positive mode. Results: Multiple calibration curves for urinary cortisol and cortisone exhibited consistent linearity and reproducibility in the range 7–828 nmol/L (0.25–30 μg/dL). Interassay CVs were 7.3–16% for mean concentrations of 6–726 nmol/L (0.2–26.3 μg/dL) for cortisol and cortisone. The detection limit was 6 nmol/L (0.2 μg/dL). Recovery of cortisol and cortisone added to urine was 97–123%. The regression equation for the LC-MS/MS (y) and HPLC (x) method for cortisol was: y = 1.11x + 0.03 μg cortisol/24 h (r2 = 0.992; n = 99). The regression equation for the LC-MS/MS (y) and immunoassay (x) methods for cortisol was: y = 0.66x − 12.1 μg cortisol/24 h (r2 = 0.67; n = 99). Conclusion: The sensitivity and specificity of the LC-MS/MS method for urinary free cortisol and cortisone offer advantages over routine immunoassays or chromatographic methods because of elimination of drug interferences, high throughput, and short chromatographic run time.

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Measurement of urinary free cortisol using liquid chromatography-tandem mass spectrometry: comparison with the urine adapted ACS:180 serum cortisol chemiluminescent immunoassay and development of a new reference range

Annals of Clinical Biochemistry International Journal of Laboratory Medicine ◽

10.1258/0004563053492775 ◽

2005 ◽

Vol 42 (2) ◽

pp. 112-118 ◽

Cited By ~ 28

Author(s):

Steven J McCann ◽

Scott Gillingwater ◽

Brian G Keevil

Keyword(s):

Mass Spectrometry ◽

Reference Range ◽

Solid Phase ◽

Serum Cortisol ◽

Urinary Free Cortisol ◽

Tandem Mass ◽

Phase Extraction ◽

Chromatography Tandem Mass Spectrometry ◽

Free Cortisol ◽

Liquid Chromatography Tandem Mass

Background: The measurement of urinary free cortisol (UFC) is commonly used in the investigation of possible Cushing's syndrome. With the recent availability of liquid chromatography-tandem mass spectrometry (LC-MS/MS) in hospital laboratories, we wanted to develop a specific UFC LC-MS/MS method and compare it with our current immunoassay method and develop a new LC-MS/MS reference range if required. Methods: A UFC LC-MS/MS method using deuterated cortisol as an internal standard was optimized using solid-phase extraction as a clean-up procedure. The multiple reaction-monitoring transitions used for the detection of cortisol and deuterated cortisol were 363.1>121 and 365.1>121.8, respectively. The method was investigated regarding precision, linearity, sensitivity, recovery and interference. UFC was measured by the in-house urine adapted ACS:180 serum cortisol immunoassay and the developed LC-MS/MS method in 110 urine samples from patients being investigated for possible Cushing's syndrome. Results: The within-batch precisions ( n=25) of the LC-MS/MS method were 7.6%, 4.5% and 3.3% at 25.0 nmol/L, 49.6 nmol/L and 344.6 nmol/L, respectively; the between-batch precisions ( n = 10) were 9.4%, 9.4% and 8.4%, respectively, at these concentrations. The method is sensitive down to 5 nmol/L and linear up to at least 1000 nmol/L. The method showed adequate cortisol recovery and no interference from the numerous drugs and steroids tested. The total run time for 20 samples, including sample preparation, was 120 min. A scatter plot of paired UFC measurements on the LC-MS/MS and the ACS:180 gave the equation: LC-MS/MS=0.408 (ACS:180)+2.65, r2 = 0.6664. The 24-h measured UFC results on 110 samples (25 men and 85 women) were positively skewed. After log transformation the data were less skewed, and following back transformation of the lower 97.5th centile, the upper limit of normal was 165 nmol/24 h. The 95th centile of the untransformed data was 146 nmol/24 h ( n=110, 25 men and 85 women). Separated by sex, the 95th centile was 152 nmol/24 h for men ( n=25) and 141 nmol/24 h for women ( n=85). Conclusions: We have developed a UFC LC-MS/MS method with a solid-phase extraction clean-up step. The method shows adequate performance and is suitable for routine laboratory use. The mixed sex ( n=110, men=25, women=85) reference range was up to 165 nmol/24 h or 146 nmol/24 h, depending on how the data are manipulated.

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Clinical utility of an ultrasensitive urinary free cortisol assay by tandem mass spectrometry

Steroids ◽

10.1016/j.steroids.2019.03.014 ◽

2019 ◽

Vol 146 ◽

pp. 65-69 ◽

Cited By ~ 1

Author(s):

Amy Luo ◽

El Taher M. El Gierari ◽

Laura M. Nally ◽

Lillian R. Sturmer ◽

Dylan Dodd ◽

...

Keyword(s):

Mass Spectrometry ◽

Tandem Mass Spectrometry ◽

Clinical Utility ◽

Urinary Free Cortisol ◽

Tandem Mass ◽

Free Cortisol

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Clinical evaluation of the liquid-chromatographic determination of urinary free cortisol.

Clinical Chemistry ◽

10.1093/clinchem/29.5.847 ◽

1983 ◽

Vol 29 (5) ◽

pp. 847-851 ◽

Cited By ~ 12

Author(s):

J Nakamura ◽

M Yakata

Keyword(s):

Normal Subjects ◽

Chromatographic Determination ◽

Urinary Free Cortisol ◽

Urinary Cortisol ◽

Liquid Chromatographic Method ◽

Free Cortisol ◽

Urinary Cortisol Excretion ◽

Liquid Chromatographic Determination ◽

Cortisol Excretion ◽

Liquid Chromatographic

Abstract We recently reported (Clin. Chem. 28: 1497-1500, 1982) a liquid-chromatographic method for quantifying free cortisol in urine. We have since evaluated the clinical utility of our method by assaying cortisol in urine from normal subjects, patients, and subjects undergoing endocrine tests. We found that, in contrast with plasma cortisol, urinary cortisol is not bound to protein. It shows some correlation with 17-hydroxycorticosteroids in urine, but is independent of creatinine excretion. The amount of cortisol excreted daily by a particular individual was found to be fairly constant during nine or 10 days. Normal values determined for 203 apparently healthy individuals were 35.8 (SD 18.7) micrograms/day, with no significant sex-related differences but a tendency for a gradual decrease of cortisol excretion with age. We also report urinary cortisol excretion by patients with pituitary-adrenal disorders and some other diseases, and the pattern of response to dexamethasone and metyrapone administration.

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Poor specificity and recovery of urinary free cortisol as determined by the Bayer ADVIA Centaur extraction method

Annals of Clinical Biochemistry International Journal of Laboratory Medicine ◽

10.1258/000456303322326506 ◽

2003 ◽

Vol 40 (5) ◽

pp. 563-565 ◽

Cited By ~ 9

Author(s):

G Gray ◽

L Shakerdi ◽

AM Wallace

Keyword(s):

Mass Spectrometry ◽

Gas Chromatography ◽

Solvent Extraction ◽

Extraction Method ◽

Urinary Free Cortisol ◽

Cross Reactivity ◽

Gas Chromatography Mass Spectrometry ◽

Sample Matrix ◽

Free Cortisol ◽

Free Serum

Background: Immunoassay methods for urinary free cortisol (UFC) lack specificity, and many procedures have not been fully evaluated for routine use. In the current study we evaluated the Bayer ADVIA Centaur extraction UFC immunoassay and compared results to those obtained by a specific gas chromatography-mass spectrometry (GC-MS) method. Results: The Bayer ADVIA Centaur cortisol assay lacked specificity. Cortisone, a steroid present in urine at concentrations similar to those of cortisol, demonstrated a cross-reactivity of 44%. In addition, the choice of matrix used to resuspend steroids after solvent extraction from urine affected recovery. Recovery was improved if the recommended urine reconstruction buffer was replaced by steroid-free serum. Conclusion: Irrespective of the sample matrix used, the Centaur method overestimates UFC, giving results up to twice those obtained by a specific GC-MS method.

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