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An FDA-Validated, Self-Cleaning Liquid Chromatography-Mass Spectrometry System for Determining Small-Molecule Drugs and Metabolites in Organoid/Organ-on-Chip Medium
S. Kogler, GM. Pedersen, F. Martínez-Ramírez, A. Aizenshtadt, M. Busek, SJK. Krauss, SR. Wilson, H. Røberg-Larsen
Jazyk angličtina Země Spojené státy americké
Typ dokumentu časopisecké články, validační studie, práce podpořená grantem
- MeSH
- chromatografie kapalinová metody MeSH
- extrakce na pevné fázi MeSH
- hmotnostní spektrometrie metody MeSH
- játra * metabolismus MeSH
- kapalinová chromatografie-hmotnostní spektrometrie MeSH
- knihovny malých molekul analýza metabolismus chemie MeSH
- laboratoř na čipu MeSH
- léčivé přípravky metabolismus analýza MeSH
- lidé MeSH
- organoidy * metabolismus cytologie MeSH
- tolbutamid metabolismus analýza MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- validační studie MeSH
As organoids and organ-on-chip (OoC) systems move toward preclinical and clinical applications, there is an increased need for method validation. Using a liquid chromatography-mass spectrometry (LC-MS)-based approach, we developed a method for measuring small-molecule drugs and metabolites in the cell medium directly sampled from liver organoids/OoC systems. The LC-MS setup was coupled to an automatic filtration and filter flush system with online solid-phase extraction (SPE), allowing for robust and automated sample cleanup/analysis. For the matrix, rich in, e.g., protein, salts, and amino acids, no preinjection sample preparation steps (protein precipitation, SPE, etc.) were necessary. The approach was demonstrated with tolbutamide and its liver metabolite, 4-hydroxytolbutamide (4HT). The method was validated for analysis of cell media of human stem cell-derived liver organoids cultured in static conditions and on a microfluidic platform according to Food and Drug Administration (FDA) guidelines with regards to selectivity, matrix effects, accuracy, precision, etc. The system allows for hundreds of injections without replacing chromatography hardware. In summary, drug/metabolite analysis of organoids/OoCs can be performed robustly with minimal sample preparation.
Citace poskytuje Crossref.org
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