High-Throughput Permeability Screening

The DMSO stock of each compound was diluted with phosphate buffer (PBS 25 mM, pH 7.4) to a final concentration of 500 µM in order to obtain the “donor”. For GI and BBB permeability, filters were coated with 10 µL of 1% w/v dodecane phosphatidylcholine solution or 5 µL of 10% w/v CHCl₃/dodecane brain polar lipid solution. The donor (150 µL) was poured over the artificial membranes on the filter plate. DMSO/PBS (300 µL, 1:1 v/v) was then added to the acceptor wells. After assembling the sandwich plates, the experiments were run for 5 h at room temperature (RT). Lastly, the amount of compound passed though the artificial membranes was determined using the UV/LC–MS method described above. Apparent permeability (Papp) and membrane retention (MR%) were determined as previously described [24 (link),25 (link)].