Quantitative multiple fragment monitoring with enhanced in-source fragmentation/annotation mass spectrometry
Analytical techniques with high sensitivity and selectivity are essential to the quantitative analysis of clinical samples. Liquid chromatography (LC) coupled to tandem mass spectrometry (MS/MS) is the gold standard in clinical chemistry. However, tandem mass spectrometers come at high capital expen...
| Autores: | , , , , , , , , , , , , , , |
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| Tipo de recurso: | artículo |
| Fecha de publicación: | 2023 |
| País: | España |
| Institución: | Universidad de Alcalá (UAH) |
| Repositorio: | e_Buah Biblioteca Digital Universidad de Alcalá |
| Idioma: | inglés |
| OAI Identifier: | oai:ebuah.uah.es:10017/64453 |
| Acceso en línea: | http://hdl.handle.net/10017/64453 https://dx.doi.org/10.1038/s41596-023-00803-0 |
| Access Level: | acceso abierto |
| Palabra clave: | Liquid chromatography Mass spectrometry Química Chemistry |
| Sumario: | Analytical techniques with high sensitivity and selectivity are essential to the quantitative analysis of clinical samples. Liquid chromatography (LC) coupled to tandem mass spectrometry (MS/MS) is the gold standard in clinical chemistry. However, tandem mass spectrometers come at high capital expenditure and maintenance costs. We recently showed that it is possible to generate very similar results using a much simpler single MS detector by performing enhanced in-source fragmentation/annotation (EISA) combined with correlated ion monitoring. Here we provide a step-by-step protocol for optimizing the analytical conditions for EISA, so anyone properly trained in LC-MS can follow and apply this technique for any given analyte. We exemplify the approach by using 2-hydroxyglutarate (2-HG) which is a clinically relevant metabolite whose D-enantiomer is considered an ?oncometabolite?, characteristic to cancers associated with mutated isocitrate dehydrogenases 1 or 2 (IDH1/2). We include procedures for determining quantitative robustness, and show results of these relating to the analysis of DL-2-HG in cells, as well as in serum samples from acute myeloid leukemia patients that contain the IDH1/2 mutation. This EISA-MS protocol is a broadly applicable and low-cost approach for the quantification of small molecules that has been developed to work well for both single quadrupole and time-of-flight mass analyzers. |
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