TABLE 1

Selected mechanism-based inactivation assay conditions, including marker substrates, concentrations, metabolites, multiple reaction monitoring transitions, and positive and negative controls Stock solutions of marker substrates were prepared in ethanol.

Isozyme
Marker Substrate
Concentration
Metabolite
MRM
Positive Control
Negative Control
μM
CYP3A4 Testosterone 200 6β-Hydroxytestosterone 305→269 Mifepristone Ketoconazole
CYP3A4 Midazolam 29 1′-Hydroxymidazolam 342→324 Mifepristone Ketoconazole
CYP2C9 Diclofenac 69 4′-Hydroxydiclofenac 312→230 Tienilic acid Sulfaphenazole
CYP2C19 S-Mephenytoin 274 4′-Hydroxymephenytoin 235→150 Ticlopidine S-(+)-N-3-Benzyl-nirvanol
CYP2D6 Bufuralol 56.5 1′-Hydroxybufuralol 278→186 Paroxetine Quinidine
CYP1A2 7-Ethoxyresorufin 10 Resorufin 214→186 Furafylline α-Naphthoflavone
Internal standard
Dextromethorphan
2.5 pg/μl
272→171

  • MRM, multiple reaction monitoring.