June 25, 2018
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.