TABLE 1

Incubation and analytical LC-MS/MS conditions for substrates in cryopreserved hepatocytes and human liver microsomes



In Vitro Conditions

LC-MS/MS Conditions
Substrate Substrate Concentration in Incubation
Incubation Time
Mass Transitions
Cone Voltage
Collision Energy

Hepatocytes
Microsomes
μM min m/z V eV
Tolbutamide 10-1000 60 60 Tolbutamide 269.05→170.15 50 15
Hydroxytolbutamide 285.05→186.1 50 17
Diclofenac (IS) 293.95→250.1 50 11
Diclofenac 0.25-200 10 20 Diclofenac 293.95→250.1 50 11
4-Hydroxydiclofenac 310→266.1 70 12
Tolbutamide (IS) 269.05→170.15 50 15
S-Warfarin 0.05-100 45 60 S-Warfarin 307.15→161.10 100 20
7-Hydroxywarfarin 323.15→177.1 130 20
Naproxen (IS) 229.15→170.05 60 15
S-Mephenytoin 5-1000 60 60 S-Mephenytoin 217.05→188.2 70 14
Hydroxymephenytoin 233.05→190.25 75 14
Sulfaphenazole (IS) 313.05→156.15 90 25
Quinidine 5-400 15 20 Quinidine 325.1→307.3 49 20
3-Hydroxyquinidine341.1→226.3 70 28
Dextromethorphan (IS) 272.15→171.35 89 40
Nifedipine 1-200 10 15 Nifedipine 347.15→315.2 50 8
Oxidized nifedipine 345.05→284.2 70 20
Dextromethorphan (IS) 272.15→171.35 89 40
Testosterone 2.5-500 10 10 Testosterone 289.15→97.2 60 20
6β-Hydroxytestosterone 305.15→269.3 80 15
Prednisone (IS) 359.15→341.4 60 10
Terfenadine 0.16-79.5 10 5 Terfenadine 427.3→436.2 95 24
Terfenadine alcohol 488.25→452.25 120 27
Fexofenadine 502.3→466.15 115 25
Azacyclonol 268.2→250.2 75 10
Metoprolol (IS) 268.2→116.2 50 18
Bufuralol 0.25-1000 15 20 Bufuralol 262.15→188.25 50 15
1-Hydroxybufuralol 278.15→186.1 70 20
Metoprolol (IS) 268.2→116.2 50 18
Dextromethorphan 0.1-1000 30 30 Dextromethorphan 272.15→171.35 89 40
Dextrorphan 258.2→199.5 70 28
3-Methoxymorphinan 258.2→213.5 70 25




Levallorphan 284.3→157.45
80
40
  • IS, internal standard.