Metabolic Clearance, Mammalian Hepatocytes

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.

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

Deliverable: Intrinsic clearance, (µL/min/10⁶ cells), t_1/2, and predicted blood extraction ratio (E_H).

Test Compound Requirement: 1-2 mg dry powder or 100-200 µL of 10 mM DMSO stock solution.