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Licensed Unlicensed Requires Authentication Published by De Gruyter May 19, 2021

Paediatric reference range for overnight urinary cortisol corrected for creatinine

Ole D. Wolthers EMAIL logo , Mark Lomax and Anne Vibeke Schmedes



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.


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).


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.


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.

Corresponding author: Ole D. Wolthers, Asthma and Allergy Clinic, Children’s Clinic Randers, Dytmaersken 9, 8900, Randers, Denmark, Phone: +45 8710 0808, Fax: +45 8643 3395, E-mail: ;

Funding source: The Foundation for Research and Development in Secondary Referral Centres


Thanks to research nurse Anne Karina Kjaer for helping with recruiting children and collecting urine samples. Thanks to children, parents and teachers at Randers Realskole for creating helpful working conditions in all involved classes and for taking part in the study.

  1. Research funding: The study was supported by a grant from “The Foundation for Research and Development in Secondary Referral Centres”. The funding organization played no role in the study design; in the collection, analysis and interpretation of the data, in the writing of the report; or in the decission to submit the report for publication.

  2. Author contributions: Ole D. Wolthers wrote the protocol, required funding, executed the protocol, took part in data processing and interpreted the data. He also wrote the paper. Mark Lomax commented on the protocol, took part in data processing and performed the statistical analysis. He also commented on the paper. Anne Vibeke Schmedes commented on the protocol, was responsible for the urine measurements and commented on the paper. All authors agree to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.

  3. Competing interests: None declared.

  4. Informed consent: Oral and written information about the study rationale were given to children and parents separately. Firstly, parents were informed of the study at parent-teacher conferences arranged by the school (Randers Realskole) and were asked for their written consent allowing their children’s participation. Thereafter, children whose parents had consented were informed about the study procedures in their classrooms and were asked for their assent to participate in the study.

  5. Ethical approval: The study did not require authorization from Ethics Committee.


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Received: 2021-01-05
Accepted: 2021-05-11
Published Online: 2021-05-19
Published in Print: 2021-08-26

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