Metabolic Clearance in Mammalian Hepatocytes
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 Concentration           1 µM
Hepatocyte Concentration           106 viable cells/mL
Percent Organic Solvent in Incubation           0.1% DMSO
Incubation Time           0, 15, 30, 60, 90, and 120 minutes
pH           7.4
Temperature           37 °C
Positive Control           Testosterone and 7-hydroxycoumarin
Analysis           LC-MS 
Number of Repeats           2

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.