A novel fully-automated method to measure steroids in serum by liquid chromatography-tandem mass spectrometry

• DOI: 10.1016/j.jmsacl.2022.12.004
• Publication Date: 2022-12-15T17:32:44Z
• Authors: François Fraissinet; Tony Pereira; Alizée Violin; Guillaume Feugray; Kalyane Bach-Ngohou; Valéry Brunel
• Keywords: Multiple reaction monitoring; 11DF; 17-hydroxyprogesterone DHEA; Lower limit of quantification MRM; SLE; delta4-androstenedione EMA; European Medicine Agency GC-MS/MS; 01 natural sciences; Automation; dehydroepiandrosterone; delta4-androstenedione; EMA; 11-deoxycortisol; UHPLC; Supported liquid extraction; Gas chromatography tandem mass spectrometry LC-MS/MS; Testosterone; DHEA; Polytetrafluoroethylene; Two dimensional ultra-high performance liquid chromatography-tandem mass spectrometry; QC; T; Ultra-high performance liquid chromatography; GC–MS/MS; 2D-UHPLC-MS/MS; 17-hydroxyprogesterone; Liquid-liquid extraction; 3. Good health; dehydroepiandrosterone D4; Lower limit of quantification; LLE; Steroids; MRM; PTFE; D4; Research Article; Gas chromatography tandem mass spectrometry; SRM; Multiple reaction monitoring QC; Radioimmunoassay; 610; Liquid chromatography tandem mass spectrometry LLE; 615; Medical technology; LC-MS/MS; R855-855.5; Solid phase extraction; LLOQ; Standard reference material; European Medicine Agency; RIA; Quality control; Liquid-liquid extraction LLOQ; 11-deoxycortisol 17OHP; 0104 chemical sciences; 17OHP; Liquid chromatography tandem mass spectrometry; SPE

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.