Off-line solid phase extraction and liquid chromatography-tandem mass spectrometry method for the quantitation of brivaracetam acid metabolites: Method validation and application to in vitro metabolism assays
| Autor: | Benoit Culot, Armel Stockis, E. Bourgogne, Sylvie Dell'Aiera, Jean-Marie Nicolas, Hugues Chanteux |
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| Rok vydání: | 2018 |
| Předmět: |
Electrospray
Metabolite Clinical Biochemistry Kidney Mass spectrometry Tandem mass spectrometry 030226 pharmacology & pharmacy Biochemistry Analytical Chemistry 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Limit of Detection Tandem Mass Spectrometry Liquid chromatography–mass spectrometry Humans Solid phase extraction Detection limit Chromatography Chemistry Hydrolysis Solid Phase Extraction Selected reaction monitoring Reproducibility of Results Cell Biology General Medicine Pyrrolidinones Linear Models Microsomes Liver 030217 neurology & neurosurgery Chromatography Liquid |
| Zdroj: | Journal of Chromatography B. 1086:138-145 |
| ISSN: | 1570-0232 |
| Popis: | Brivaracetam (BRV) is a new high affinity synaptic vesicle protein 2A ligand recently approved for adults with partial-onset seizures. As a support to in vitro metabolism assays, a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method coupled to off-line solid phase extraction (SPE) was developed to quantify BRV acid metabolites, that is, BRV-AC (carboxylic derivative derived from BRV hydrolysis) and BRV-OHAC (corresponding to hydroxylated BRV-AC). The method was validated for various incubates (liver and kidney tissue homogenates and blood, all from humans) and applied to in vitro metabolism assays. The analytes were isolated from buffered samples using ISOLUTE C8 96-well SPE plates. Chromatographic separation was achieved on a Waters Atlantis T3 C18 analytical column (2.1 mm × 50 mm, 5 μm) with detection accomplished using a Waters Premier tandem mass spectrometer in positive ion electrospray and multiple reaction monitoring (MRM) mode. The standard curves, which ranged from 1.00 to 200 ng/mL for BRV-AC, BRV-OHAC, were fitted to a 1/x2 weighted linear regression model. The intra-assay precision and inter-assay precision (expressed as coefficient of variation -%CV) were |
| Databáze: | OpenAIRE |
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