Enzyme | Substrate Concentration | Protein | Incubation Time | Ionization Mode | Mass Transition of Metabolite | Metabolite Monitored (Internal Standard) | |||
---|---|---|---|---|---|---|---|---|---|
HLM | rUGT | HLM | rUGT | HLM | rUGT | ||||
µM | mg/ml | min | |||||||
UGT1A1 | 9 | 9 | 0.1 | 0.1 | 5 | 5 | Negative | 447 / 271 | 17β-Estradiol-3-glucuronide (17β-Estradiol 3-glucuronide-d5) |
UGT1A3 | 20 | 20 | 0.1 | 0.1 | 10 | 5 | Negative | 567 / 391 | Chenodeoxycholic acid 24-O-glucuronide (chenodeoxycholic acid 24-O-glucuronide-d5) |
UGT1A4 | 12 | 12 | 0.1 | 0.1 | 5 | 5 | Positive | 584 / 408 | Trifluoperazine N-glucuronide (prochlorperazine N-glucuronide) |
UGT1A6 | 1 | 1 | 0.005 | 0.005 | 5 | 5 | Negative | 319 / 143 | 1-Naphthol O-glucuronide (1-naphthol O-glucuronide-d7) |
UGT1A9 | 20 | 5 | 0.1 | 0.1 | 5 | 5 | Negative | 353 / 177 | Propofol O-glucuronide (propofol O-glucuronide-d17) |
UGT2B7 | 400 | 400 | 0.1 | 0.1 | 5 | 5 | Positive | 462 / 286 | Morphine 3-O-glucuronide (morphine 3-O-glucuronide-d3) |
UGT2B10 | 3 | 3 | 0.1 | 0.1 | 10 | 5 | Positive | 377 / 201 | Levomedetomidine N-glucuronide (1ʹ-hydroxymidazolam-d4) |
UGT2B15 | 50 | 20 | 0.1 | 0.1 | 10 | 5 | Positive | 463 / 286 | S-Oxazepam O-glucuronide (S-oxazepam O-glucuronide-d5) |
UGT2B17 | 5 | 2.5 | 0.1 | 0.1 | 10 | 5 | Positive | 465 / 289 | Testosterone 17β-O-glucuronide (testosterone 17β-O-glucuronide-d5) |
UGT1A7 | NA | 14 | NA | 0.1 | NA | 5 | Positive | 353 / 177 | 4-Methylumbelliferone glucuronide (7-hydroxycoumarin-d5) |
UGT1A8 | 730 | 0.1 | 5 | ||||||
UGT1A10 | 31 | 0.1 | 30 | ||||||
UGT2B4 | 1200 | 0.25 | 15 |
HLM, human liver microsomes; rUGT, recombinant UDP-glucuronosyltransferase; NA, not applicable.