October 3, 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 Concentration1 µM
S9 Protein Concentration1.25 mg/mL
CofactorsNADPH, UDPGA, and PAPS
Percent Organic Solvent in Incubation0.5% acetonitrile
Incubation Time0, 5, 10, 15, 30, and 45 minutes
pH7.4
Temperature37 °C
Positive ControlTestosterone and 7-hydroxycoumarin
AnalysisLC-MS (LC-MS/MS if necessary)
Number of Repeats2

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.

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