October 3, 2018

Assay System:

  • Cryopreserved mammalian hepatocytes in suspension.
  • Human hepatocytes 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 hepatocytes.

Assay Conditions:

Test Compound Concentration1 µM
Hepatocyte Concentration106 viable cells/mL
Percent Organic Solvent in Incubation0.1% DMSO
Incubation Time0, 15, 30, 60, 90, and 120 minutes
pH7.4
Temperature37 °C
Positive ControlTestosterone and 7-hydroxycoumarin
AnalysisLC-MS
Number of Repeats2

Deliverable: Intrinsic clearance, (µL/min/106 cells), 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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