Free cortisol method comparison: Ultrafiltation, equilibrium dialysis, tracer dilution, tandem mass spectrometry and calculated free cortisol
Abstract
Background Free cortisol (FC) can be calculated from measurements of total cortisol and binding proteins or measured after mechanical separation of unbound and bound fractions by equilibrium dialysis or ultrafiltration. FC can then be measured indirectly by 3H-cortisol dilution or directly by immunologic or tandem mass spectrometry assays. Methods We compared FC measured with ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC MS/MS) with 3H-cortisol dilution in ultrafiltrates and dialysates and also with calculated FC (Coolens equation). An adult FC reference interval was established. Results The UHPLC MS/MS and 3H-cortisol dilution methods were non-linearly related (Cusum linearity test p Conclusions In view of the non-linearity between measuring techniques and the variability of reported reference ranges, care should be exercised in adopting a reference range. The ultrafiltration UHPLC MS/MS method we described is robust and suitable for use in a routine laboratory.
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Indirect ED is subject to limitations (tracer instability, tracer purity, need of a dilution step) that introduce added total error to free steroid measurement compared to direct ED.
“Indirect ED, as compared to direct ED, is subject to several limitations, such as tracer instability, tracer purity and need of a dilution step, that all introduce possible sources of added total error to the free steroid measurement [199, 200].”
From Role of sex hormone-binding globulin in the free hormone hypothesis and the relevance of free testosterone in androgen physiology (2022) · cites this paper as [199]
LC-MS/MS coupled ultrafiltration (UF) is a comparable alternative to ED and, when properly validated, can display equal performance to ED.
“Briefly, LC–MS/MS coupled ultrafiltration (UF) constitutes a comparable alternative to ED.”
From Role of sex hormone-binding globulin in the free hormone hypothesis and the relevance of free testosterone in androgen physiology (2022) · cites this paper as [199]
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- Authors
- Pretorius, C. J., Galligan, J. P., McWhinney, B. C., Briscoe, S. E., & Ungerer, J. P. J.
- Affiliations
- Queensland Health (AU)
- Corresponding author
- Pretorius, Carel J.
- Venue
- Clinica Chimica Acta
- Publisher
- Elsevier BV
- Published
- 2011-05
- Volume
- 412
- Issue
- 11-12
- Pages
- 1043-1047
- ISSN
- 0009-8981 (ISSN-L)
- OpenAlex
- W2028414960
- Cited by
- 35
- References
- 4
- Citations by year
- 2024: 1 · 2023: 1 · 2022: 1 · 2021: 2 · 2020: 2 · 2019: 1 · 2018: 4 · 2017: 2 · 2016: 5 · 2015: 3 · 2014: 7 · 2013: 4 · 2012: 1
- FWCI
- 3.41
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Clinica Chimica Acta (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Adrenal Hormones and Disorders
- Health Sciences › Medicine › Physiology › Clinical Laboratory Practices and Quality Control
- Life Sciences › Neuroscience › Behavioral Neuroscience › Stress Responses and Cortisol
- Keywords
- free cortisol, method comparison, ultrafiltration, equilibrium dialysis, tandem mass spectrometry, reference values, method linearity, cortisol-binding protein, nonspecific adsorption
- MeSH terms
- Adult, Blood Chemical Analysis, Chromatography, High Pressure Liquid, Female, Humans, Hydrocortisone, Male, Middle Aged, Reference Values, Ultrafiltration, Tandem Mass Spectrometry, Young Adult
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