Caco-2 Standard Test Method
Papp Value Determination

TESTING PARAMETERS

• Compounds cold or radiolabeled
• Two concentrations per compound tested
• Three replicates per compound per dose
• Samples taken Apical to Basolateral side at 0, 15, 30, 60, 90, and 120 minutes. Samples taken on the Apical side at 0 and 120 minutes.
• Data tables and graphs for each compound electronically transferred to the client
• If Basolateral to Apical assessment is performed, samples taken at the same time points as specified above.

TEST CONDITIONS

• Caco-2 cell membranes prepared with 21 to 28 days' incubation in 6-well SnapWell® plates
• Each set of membranes tested for membrane integrity and function by determining the permeability coefficients for mannitol and phenytoin
• Solubility of each compound determined in water; if necessary, also in DMSO and ethanol
• Two test compound concentrations
• Three replicates per concentration
• Two sampling time points from the donor-side and six sampling time points from the receiver-side
• Sample concentration analysis performed by LCMS or other appropriate method

Methods may be modified to meet a particular test format.