Experimental conditions for measurement of P450 and UGT activity in HLMs and CHHs
| Enzyme | Marker Activity | Substrate Concentration | HLMs | CHHs | Incubation Time in HLMs | Incubation Time in CHHs | Mass Transition Monitored | Ionization Mode | Internal Standard |
|---|---|---|---|---|---|---|---|---|---|
| μM | mg/ml | million cells/ml | min | min | m/z | ||||
| CYP1A2 | Phenacetin O-deethylation | 40 | 0.1 | 0.5 | 5 | 10 | 150.0/107.0 | Negative | d4-Acetaminophen |
| CYP2B6 | Bupropion hydroxylation | 50 | 0.1 | 0.5 | 5 | 10 | 256.1/238.0 | Positive | d6-Hydroxybupropion |
| CYP2C8 | Amodiaquine N-dealkylation | 1.5 (7 for CHHs) | 0.0125 | 0.5 | 5 | 10 | 328.2/283.1 | Positive | d5-N-Desethylamodiaquine |
| CYP2C9 | Diclofenac 4′-hydroxylation | 6 | 0.1 | 0.5 | 5 | 10 | 310.0/266.0 | Negative | d4-4′-Hydroxydiclofenac |
| CYP2C19 | S-Mephenytoin 4′-hydroxylation | 40 | 0.1 | 0.5 | 5 | 10 | 233.0/190.0 | Negative | d3-4′-Hydroxymephenytoin |
| CYP2D6 | Dextromethorphan O-demethylation | 7.5 | 0.1 | 0.5 | 5 | 10 | 258.0/157.1 | Positive | d3-Dextrorphan |
| CYP3A4/5 | Midazolam 1′-hydroxylation | 4 | 0.1 | 0.5 | 5 | 10 | 342.0/324.0 | Positive | d4-1′-Hydroxymidazolam |
| UGT1A1 | Estradiol 3-O-glucuronidation | 9 | 0.1 | NA | 5 | NA | 4471/271.2 | Negative | d5-Estradiol 3-O-glucuronide |
| UGT1A3 | Chenodeoxycholic acid 24-O-glucuronidation | 20 | 0.1 | NA | 10 | NA | 567.2/391.3 | Negative | d5-Chenodeoxycholic acid 24-O-glucuronide |
| UGT1A4 | Trifluoperazine glucuronidation | 12 | 0.1 | NA | 5 | NA | 584.0/408.0 | Positive | Prochlorperazine glucuronide |
| UGT1A6 | 1-Naphthol glucuronidation | 1 | 0.0125 | NA | 5 | NA | 319.1/143.1 | Negative | d7-Naphthol glucuronide |
| UGT1A9 | Propofol glucuronidation | 20 | 0.1 | NA | 5 | NA | 353.2/177.1 | Negative | d17-Propofol glucuronide |
| UGT2B7 | Morphine 3-O-glucuronidation | 400 | 0.1 | NA | 5 | NA | 462.0/286.0 | Positive | d3-Morphine 3-O-glucuronide |
| UGT2B10 | Levomedetomidine glucuronidation | 7 | 0.1 | NA | 10 | NA | 377.0/201.0 | Positive | d4-1′-Hydroxymidazolam |
| UGT2B15 | S-Oxazepam glucuronidation | 50 | 0.1 | NA | 10 | NA | 463.0/286.4 | Positive | d5-S-Oxazepam glucuronide |
| UGT2B17 | Testosterone 17-O-glucuronidation | 5 | 0.1 | NA | 10 | NA | 465.1/289.0 | Positive | d5-Testosterone 17-O-glucuronide |
| UGT | 4-Methylumbelliferone glucuronidation | 70 | NA | 0.5 | NA | 10 | 352.9/177.1 | Positive | d5-7-Hydroxycoumarin glucuronide |
NA, not applicable.