Semi-automated serum steroid profiling with tandem mass spectrometry
Open accessAbstract
Objectives: Highly selective and sensitive multi-analyte methods for the analysis of steroids are attractive for the diagnosis of endocrine diseases. Commercially available kits are increasingly used for this purpose. These methods involve laborious solid phase extraction, and the respective panels of target analytes are incomplete. We wanted to investigate whether an improvement of kit solutions is possible by introducing automated on-line solid phase extraction (SPE) and combining originally separate analyte panels. Methods: Sample preparation was performed using automated on-line SPE on a high-pressure stable extraction column. Chromatographic separation, including isobaric compounds, was achieved using a 0.25 mM ammonium fluoride-methanol gradient on a small particle size biphenyl column. Standard compounds and internal standard mixtures of two panels of a commercially available kit were combined to achieve an optimized and straightforward detection of 15 endogenous steroids. Validation was performed according to the European Medicines Agency (EMA) guidelines with slight modifications. Results: Validation was successfully performed for all steroids over a clinically relevant calibration range. Deviations of intra- and inter-assay accuracy and precision results passed the criteria and no relevant matrix effects were detected due to highly effective sample preparation. External quality assessment samples showed the applicability as a routine diagnostic method, which was affirmed by the analyses of anonymized clinical samples. Conclusions: It was found possible to complement a commercially available kit for quantitative serum steroid profiling based on isotope dilution LC-MS/MS by implementing automated on-line SPE, thereby improving the practicality and robustness of the measurement procedure.
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Several sensitive and robust LC-MS/MS methods for multiple steroids have been developed in recent years and many methods meet the requirements for clinical applications.
“Several sensitive and robust LC-MS/MS methods for multiple steroids have been developed in recent years and many methods meet the re quirements for clinical applications [5 -16].”
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Cited by
Sorted by citations
Frederiksen, H., Johannsen, T. H., Andersen, S. E., Petersen, J. H., Busch, A. S., Ljubicic, M. L., Fischer, M. B., Upners, E. N., Hagen, C. P., Main, K. M., et al. (2024) · Clinica Chimica Acta · doi:10.1016/j.cca.2024.119852 · Cited by 46 · see citing works
- Authors
- Rakete, S., Schubert, T., & Vogeser, M.
- Affiliations
- Institut für Laboratoriumsmedizin (DE)
- Corresponding author
- Vogeser, Michael
- ORCID
- Vogeser, Michael
- Venue
- Journal of Mass Spectrometry and Advances in the Clinical Lab
- Publisher
- Stanford University Press
- Published
- 2023-01
- Volume
- 27
- Pages
- 40-48
- ISSN
- 2667-145X (ISSN-L)
- PMC
- PMC9813517
- OpenAlex
- W4311748034
- Cited by
- 10
- References
- 35
- Citations by year
- 2026: 4 · 2025: 2 · 2024: 1 · 2023: 3
- FWCI
- 0.49
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Journal of Mass Spectrometry and Advances in the Clinical Lab (publishedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Metabolomics and Mass Spectrometry Studies
- Physical Sciences › Chemistry › Analytical Chemistry › Analytical chemistry methods development
- Physical Sciences › Chemistry › Spectroscopy › Analytical Chemistry and Chromatography
- Keywords
- tandem mass spectrometry, isotope dilution LC-MS/MS, endocrine disease diagnosis, method validation, external quality assessment
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