Metabolic Clearance, Liver S9 Fractions
Assay System:
- Mammalian liver S9 fractions.
- Human S9 fractions are pooled from 10 donors (mixed gender). All other species are male.
- Metabolic clearance determined by measuring percent remaining of test compound at multiple time points during incubation with liver S9 fractions.
Assay Conditions:
| Test Compound Concentration |
1 µM |
| S9 Protein Concentration |
1.25 mg/mL |
| Cofactors |
NADPH, UDPGA, and PAPS |
| Percent Organic Solvent in Incubation |
0.5% acetonitrile |
| Incubation Time |
0, 5, 10, 15, 30, and 45 minutes |
| pH |
7.4 |
| Temperature |
37 °C |
| Positive Control |
Testosterone and 7-hydroxycoumarin |
| Analysis |
LC-MS (LC-MS/MS if necessary) |
| Number of Repeats |
2 |
Deliverable: Intrinsic clearance, t1/2, and predicted blood extraction ratio (EH).
Test Compound Requirement: 1-2 mg dry powder or 100-200 µL of 10 mM DMSO stock solution.
