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Determination of reference intervals for urinary steroid profiling using a newly validated GC-MS/MS method

  • Wilhelmina H.A. de Jong EMAIL logo , Edward Buitenwerf , Alle T. Pranger , Ineke J. Riphagen ORCID logo , Bruce H.R. Wolffenbuttel , Michiel N. Kerstens and Ido P. Kema
Published/Copyright: June 9, 2017
Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 56 Issue 1

Abstract

Background:

Urinary steroid profiling (USP) is a powerful diagnostic tool to asses disorders of steroidogenesis. Pre-analytical factors such as age, sex and use of oral contraceptive pills (OCP) may affect steroid hormone synthesis and metabolism. In general, USP reference intervals are not adjusted for these variables. In this study we aimed to establish such reference intervals using a newly-developed and validated gas chromatography with tandem mass spectrometry detection method (GC-MS/MS).

Methods:

Two hundred and forty healthy subjects aged 20–79 years, stratified into six consecutive decade groups each containing 20 males and 20 females, were included. None of the subjects used medications. In addition, 40 women aged 20–39 years using OCP were selected. A GC-MS/MS assay, using hydrolysis, solid phase extraction and double derivatization, was extensively validated and applied for determining USP reference intervals.

Results:

Androgen metabolite excretion declined with age in both men and women. Cortisol metabolite excretion remained constant during life in both sexes but increased in women 70–79 years of age. Progesterone metabolite excretion peaked in 30–39-year-old women and declined afterwards. Women using OCP had lower excretions of androgen metabolites, progesterone metabolites and cortisol metabolites. Method validation results met prerequisites and revealed the robustness of the GC-MS/MS method.

Conclusions:

We developed a new GC-MS/MS method for USP which is applicable for high throughput analysis. Widely applicable age and sex specific reference intervals for 33 metabolites and their diagnostic ratios have been defined. In addition to age and gender, USP reference intervals should be adjusted for OCP use.

Keywords: GC-MS/MS; oral contraceptive pills; reference intervals; urinary steroid profile
Corresponding author: Wilhelmina H.A. de Jong, PhD, Department of Laboratory Medicine, University of Groningen, University Medical Center Groningen, Hanzeplein 1, EA61, PO-Box 30.001, 9700 RB Groningen, The Netherlands, Phone: +0031-50-3617015, Fax: +050-3611746

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: None declared.

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

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Supplemental Material:

The online version of this article (https://doi.org/10.1515/cclm-2016-1072) offers supplementary material, available to authorized users.

Received: 2016-11-24
Accepted: 2017-4-20
Published Online: 2017-6-9
Published in Print: 2017-11-27

©2018 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2016-1072
Keywords for this article
GC-MS/MS; oral contraceptive pills; reference intervals; urinary steroid profile

Articles in the same Issue

  1. Frontmatter
  2. Editorial
  3. Method comparison – a practical approach based on error identification
  4. Review
  5. Neutrophil gelatinase-associated lipocalin as a risk marker in cardiovascular disease
  6. Mini Reviews
  7. α-Defensin point-of-care test for diagnosis of prosthetic joint infections: neglected role of laboratory and clinical pathologists
  8. The diagnostic accuracy of biomarkers for diagnosis of primary biliary cholangitis (PBC) in anti-mitochondrial antibody (AMA)-negative PBC patients: a review of literature
  9. Opinion Paper
  10. New issues on measurement of B-type natriuretic peptides
  11. Genetics and Molecular Diagnostics
  12. The SEeMORE strategy: single-tube electrophoresis analysis-based genotyping to detect monogenic diseases rapidly and effectively from conception until birth
  13. General Clinical Chemistry and Laboratory Medicine
  14. Determination of serum calcium levels by 42Ca isotope dilution inductively coupled plasma mass spectrometry
  15. The effects of dry ice exposure on plasma pH and coagulation analyses
  16. Placental protein-13 (PP13) in combination with PAPP-A and free leptin index (fLI) in first trimester maternal serum screening for severe and early preeclampsia
  17. Circulating CD89-IgA complex does not predict deterioration of kidney function in Korean patients with IgA nephropathy
  18. Performance analysis of automated evaluation of Crithidia luciliae-based indirect immunofluorescence tests in a routine setting – strengths and weaknesses
  19. Performance of automated digital cell imaging analyzer Sysmex DI-60
  20. Reference Values and Biological Variations
  21. Determination of reference intervals for urinary steroid profiling using a newly validated GC-MS/MS method
  22. Reference intervals and longitudinal changes in copeptin and MR-proADM concentrations during pregnancy
  23. Definition of the upper reference limit of glycated albumin in blood donors from Italy
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  37. Reply to: Analytical evaluation of the performances of Diazyme and BRAHMS procalcitonin applied to Roche Cobas in comparison with BRAHMS PCT-sensitive Kryptor
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