A novel fully-automated method to measure steroids in serum by liquid chromatography-tandem mass spectrometry.
11DF, 11-deoxycortisol
17OHP, 17-hydroxyprogesterone
2D-UHPLC-MS/MS, Two dimensional ultra-high performance liquid chromatography-tandem mass spectrometry
Automation
D4, delta4-androstenedione
DHEA, dehydroepiandrosterone
EMA, European Medicine Agency
GC–MS/MS, Gas chromatography tandem mass spectrometry
LC-MS/MS, Liquid chromatography tandem mass spectrometry
LLE, Liquid-liquid extraction
LLOQ, Lower limit of quantification
Liquid chromatography tandem mass spectrometry
MRM, Multiple reaction monitoring
PTFE, Polytetrafluoroethylene
QC, Quality control
RIA, Radioimmunoassay
Radioimmunoassay
SLE, Supported liquid extraction
SPE, Solid phase extraction
SRM, Standard reference material
Steroids
T, Testosterone
Testosterone
UHPLC, Ultra-high performance liquid chromatography
Journal
Journal of mass spectrometry and advances in the clinical lab
ISSN: 2667-145X
Titre abrégé: J Mass Spectrom Adv Clin Lab
Pays: Netherlands
ID NLM: 101776811
Informations de publication
Date de publication:
Jan 2023
Jan 2023
Historique:
received:
29
03
2022
revised:
24
11
2022
accepted:
12
12
2022
entrez:
3
1
2023
pubmed:
4
1
2023
medline:
4
1
2023
Statut:
epublish
Résumé
Steroids play a key role in numerous physiological processes. Steroid determination is a useful tool to explore various endocrine diseases. Because of its specificity, mass spectrometry is considered to be a reference method for the determination of steroids in serum compared to radioimmunoassay. This technology could progress towards more automation for the optimal organization of clinical laboratories and ultimately for the benefit of patients. A fully automated ultra-high-performance liquid chromatography-tandem mass spectrometry method was developed and fully validated to determine five steroids in serum. Sample preparation was based on protein precipitation with filtration followed by online solid phase extraction. Chromatographic separation was performed using a biphenyl stationary phase. The method was successfully validated according to European Medicine Agency guidelines. Coefficients of variation did not exceed, respectively, 8.4% and 8.1% for intra- and inter-assay precision. Method comparison with radioimmunoassay showed a proportional bias for all compounds, except for testosterone in men. Comparison with another LC-MS/MS method demonstrated acceptable concordance for all steroids, although a small bias was observed for androstenedione. The novelty of this method is that it has been fully automated. Automation provides benefits in traceability and allows significant savings in cost and time.
Sections du résumé
Background
UNASSIGNED
Steroids play a key role in numerous physiological processes. Steroid determination is a useful tool to explore various endocrine diseases. Because of its specificity, mass spectrometry is considered to be a reference method for the determination of steroids in serum compared to radioimmunoassay. This technology could progress towards more automation for the optimal organization of clinical laboratories and ultimately for the benefit of patients.
Methods
UNASSIGNED
A fully automated ultra-high-performance liquid chromatography-tandem mass spectrometry method was developed and fully validated to determine five steroids in serum. Sample preparation was based on protein precipitation with filtration followed by online solid phase extraction. Chromatographic separation was performed using a biphenyl stationary phase.
Results
UNASSIGNED
The method was successfully validated according to European Medicine Agency guidelines. Coefficients of variation did not exceed, respectively, 8.4% and 8.1% for intra- and inter-assay precision. Method comparison with radioimmunoassay showed a proportional bias for all compounds, except for testosterone in men. Comparison with another LC-MS/MS method demonstrated acceptable concordance for all steroids, although a small bias was observed for androstenedione.
Conclusion
UNASSIGNED
The novelty of this method is that it has been fully automated. Automation provides benefits in traceability and allows significant savings in cost and time.
Identifiants
pubmed: 36593910
doi: 10.1016/j.jmsacl.2022.12.004
pii: S2667-145X(22)00046-3
pmc: PMC9804132
doi:
Types de publication
Journal Article
Langues
eng
Pagination
24-32Informations de copyright
© 2022 THE AUTHORS.
Déclaration de conflit d'intérêts
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
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