Table 2. CYP450 Inhibition Assay Conditions.
CYP450 isoform | substrate | [S] (μM) | [HLM] (mg/mL) | incubation time (min) | metabolite (mass transition, amua) | internal standard (mass transition, amua) |
---|---|---|---|---|---|---|
CYP1A2 | tacrine | 1 | 0.2 | 10 | 1-hydroxytacrine (215 → 182) | bucetin (224 → 136) |
CYP2B6 | bupropion | 80 | 0.2 | 20 | hydroxybupropion (256 → 139) | hydroxybupropion-d6 (262 → 244) |
CYP2C8 | amodiaquine | 1.5 | 0.02 | 5 | desethylamodiaquine (330 → 285) | desethylamodiaquine-d3 (333 → 285) |
CYP2C9 | diclofenac | 5 | 0.05 | 5 | 4′-hydroxydiclofenac (312 → 268) | 4′-hydroxydiclofenac-13C6 (316 → 272) |
CYP2C19 | (S)-mephenytoin | 40 | 0.3 | 10 | 4′-hydroxy-(S)-mephenytoin (235 → 150) | 4′-hydroxy-(S)-mephenytoin-d3 (238 → 150) |
CYP2D6 | dextromethorphan | 5 | 0.1 | 5 | dextrorphan (258 → 157) | dextrorphan-d3 (261 → 157) |
CYP3A4 | midazolam | 3 | 0.02 | 5 | 1′-hydroxymidazolam (342 → 203) | 1′-hydroxymidazolam-13C3 (347 → 208) |
CYP3A4 | testosterone | 50 | 0.05 | 10 | 6β-hydroxytestosterone (305 → 269) | 6β-hydroxytestosterone-d7 (312 → 276) |
Atomic mass units.