Semi-automated serum steroid profiling with tandem mass spectrometry
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...
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| Format: | Article |
| Language: | English |
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Elsevier
2023-01-01
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| Series: | Journal of Mass Spectrometry and Advances in the Clinical Lab |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2667145X22000487 |
| _version_ | 1828046574919352320 |
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| author | Sophie Rakete Tom Schubert Michael Vogeser |
| author_facet | Sophie Rakete Tom Schubert Michael Vogeser |
| author_sort | Sophie Rakete |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-10T18:28:52Z |
| format | Article |
| id | doaj.art-722922828525490cbd2bb45704c98c12 |
| institution | Directory Open Access Journal |
| issn | 2667-145X |
| language | English |
| last_indexed | 2024-04-10T18:28:52Z |
| publishDate | 2023-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Mass Spectrometry and Advances in the Clinical Lab |
| spelling | doaj.art-722922828525490cbd2bb45704c98c122023-02-02T04:50:51ZengElsevierJournal of Mass Spectrometry and Advances in the Clinical Lab2667-145X2023-01-01274048Semi-automated serum steroid profiling with tandem mass spectrometrySophie Rakete0Tom Schubert1Michael Vogeser2Institute of Laboratory Medicine, University Hospital, LMU Munich, GermanyInstitute of Laboratory Medicine, University Hospital, LMU Munich, GermanyCorresponding author at: Institute of Laboratory Medicine, University Hospital, LMU Munich, Marchioninistrasse 15, 81377 Munich, Germany.; Institute of Laboratory Medicine, University Hospital, LMU Munich, GermanyObjectives: 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.http://www.sciencedirect.com/science/article/pii/S2667145X22000487Serum steroid profilingOn-line solid phase extraction (SPE)Stable-isotope dilution liquid chromatography-tandem mass spectrometry (SID LC-MS/MS)Robustness |
| spellingShingle | Sophie Rakete Tom Schubert Michael Vogeser Semi-automated serum steroid profiling with tandem mass spectrometry Journal of Mass Spectrometry and Advances in the Clinical Lab Serum steroid profiling On-line solid phase extraction (SPE) Stable-isotope dilution liquid chromatography-tandem mass spectrometry (SID LC-MS/MS) Robustness |
| title | Semi-automated serum steroid profiling with tandem mass spectrometry |
| title_full | Semi-automated serum steroid profiling with tandem mass spectrometry |
| title_fullStr | Semi-automated serum steroid profiling with tandem mass spectrometry |
| title_full_unstemmed | Semi-automated serum steroid profiling with tandem mass spectrometry |
| title_short | Semi-automated serum steroid profiling with tandem mass spectrometry |
| title_sort | semi automated serum steroid profiling with tandem mass spectrometry |
| topic | Serum steroid profiling On-line solid phase extraction (SPE) Stable-isotope dilution liquid chromatography-tandem mass spectrometry (SID LC-MS/MS) Robustness |
| url | http://www.sciencedirect.com/science/article/pii/S2667145X22000487 |
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