High-throughput cytochrome P450 (CYP) inhibition screening via cassette probe-dosing strategy: III. Validation of a direct injection/on-line guard cartridge extraction−tandem mass spectrometry method for CYP2C19 inhibition evaluation
| Autor: | Philip Teitelbaum, Lisa Magis, Hai-Zhi Bu, Kim Knuth |
|---|---|
| Rok vydání: | 2001 |
| Předmět: |
Chromatography
Formic acid Metabolite Clinical Biochemistry Pharmaceutical Science Mass spectrometry Tandem mass spectrometry High-performance liquid chromatography Mass Spectrometry Mixed Function Oxygenases Analytical Chemistry Cytochrome P-450 CYP2C19 Cartridge chemistry.chemical_compound chemistry Calibration Drug Discovery Microsome Cytochrome P-450 Enzyme Inhibitors Sample preparation Aryl Hydrocarbon Hydroxylases Spectroscopy |
| Zdroj: | Journal of Pharmaceutical and Biomedical Analysis. 25:437-442 |
| ISSN: | 0731-7085 |
| DOI: | 10.1016/s0731-7085(00)00518-5 |
| Popis: | A new direct injection/on-line guard cartridge extraction-tandem mass spectrometry (DI/GCE-MS-MS) method has been validated for high-throughput evaluation of cytochrome P450 (CYP) 2C19 inhibition potential using human hepatic microsomes and 96-well microtiter plates. Microsomal incubations were quenched with formic acid, centrifuged, and the resulting supernatants were injected for DI/GCE-MS-MS analysis. Due to the use of a C(18) guard cartridge (4x2.0 mm I.D.), this method exhibits several advantages such as little sample preparation, rapid on-line extraction, short analysis time (2.5 min), and minimal source contamination and performance deterioration. The DI/GCE-MS-MS method demonstrates an inter-day accuracy ranged from 0.3 to 2.4% with precision ranging from 2.0 to 3.0% for the quantification of 4-hydroxymephenytoin, a marker metabolite of S-mephenytoin mediated by CYP2C19, in microsomal incubations. The CYP2C19 inhibition assay has been validated using tranylcypromine as a known inhibitor of the isoform. The IC(50) value (43.5 microM) measured by the new method is in agreement with a reported literature value (approximately 30 microM). |
| Databáze: | OpenAIRE |
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