| Autor: |
Gadara D; RECETOX Centre, Faculty of Science, Masaryk University, Brno 625 00, Czech Republic., Berka V; RECETOX Centre, Faculty of Science, Masaryk University, Brno 625 00, Czech Republic., Spacil Z; RECETOX Centre, Faculty of Science, Masaryk University, Brno 625 00, Czech Republic. |
| Jazyk: |
angličtina |
| Zdroj: |
Analytical chemistry [Anal Chem] 2024 Jan 09; Vol. 96 (1), pp. 59-66. Date of Electronic Publication: 2023 Dec 19. |
| DOI: |
10.1021/acs.analchem.3c02652 |
| Abstrakt: |
Microflow liquid chromatography interfaced with mass spectrometry (μLC-MS/MS) is increasingly applied for high-throughput profiling of biological samples and has been proven to have an acceptable trade-off between sensitivity and reproducibility. However, lipidomics applications are scarce. We optimized a μLC-MS/MS system utilizing a 1 mm inner diameter × 100 mm column coupled to a triple quadrupole mass spectrometer to establish a sensitive, high-throughput, and robust single-shot lipidomics workflow. Compared to conventional lipidomics methods, we achieve a ∼4-fold increase in response, facilitating quantification of 351 lipid species from a single iPSC-derived cerebral organoid during a 15 min LC-MS analysis. Consecutively, we injected 303 samples over ∼75 h to prove the robustness and reproducibility of the microflow separation. As a proof of concept, μLC-MS/MS analysis of Alzheimer's disease patient-derived iPSC cerebral organoid reveals differential lipid metabolism depending on APOE phenotype (E3/3 vs E4/4). Microflow separation proves to be an environmentally friendly and cost-effective method as it reduces the consumption of harmful solvents. Also, the data demonstrate robust, in-depth, high-throughput performance to enable routine clinical or biomedical applications. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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