Incubation conditions for P450 inhibition assays
| Enzyme | Activity | Substrate Conc. | Incubation Time | HLM Conc. | Detection |
|---|---|---|---|---|---|
| μM | min | mg/ml | |||
| CYP1A2 | Ethoxyresorufin O-dealkylase | 5 | Suitable | 0.25 | Fluorescence |
| CYP2B6 | Bupropion 2-hydroxylase | 70 | 10 | 0.2 | Mass spectrometry |
| CYP2C8 | Paclitaxel 6α-hydroxylase | 10 | 10 | 0.05 | Mass spectrometry |
| CYP2C9 | Diclofenac 4′-hydroxylase | 5 | 15 | 0.2 | Mass spectrometry |
| CYP2C19 | (S)-Mephenytoin 4′-hydroxylase | 200 | 30 | 2 | Radioactivity |
| CYP2D6 | Dextromethorphan O-demethylase | 5 | 15 | 0.2 | Mass spectrometry |
| CYP2E1 | Chlorzoxazone 6-hydroxylase | 100 | 30 | 2 | Radioactivity |
| CYP3A4 | Midazolam 1′- and 4-hydroxylase | 3 | 5 | 0.2 | Mass spectrometry |
| Testosterone 6β-hydroxylase | 150 | 15 | 2 | Radioactivity |