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P2038
A High-Throughput Permeability Screening Methodology Utilizing a Novel Artificial PAMPA Membrane & LC/MS/MS Detection
Presenter Grace Poon, Syndexa Pharmaceuticals, USA
Additional Authors: Xiaoxi (Kevin)
PAMPA (Parallel Artificial Membrane Permeability Assay) consists of an immobilized lipid membrane that measures permeability of drugs across an artificial membrane. It is a well established in vitro assay in 96-well plate format, hence amenable for automation. Recently, BD Biosciences has developed a 96-well pre-coated PAMPA plate in which the microporous filters are pre-coated with a new artificial membrane – structured layers of phospholipids that mimic the biological membrane. The pre-coated PAMPA plate is stable for a prolonged period of time when stored at -20º C. The new artificial membrane produces correct prediction for the compounds under-predicted by traditional PAMPA.
In this study a set of standard compounds were used to validate the pre-coated PAMPA plates. Analytes (in triplicate) at 25 uM concentration were spiked onto the donor side, the 96-well filter plate pre-coated with lipids was used as the permeation acceptor. Inc.ubation was performed at room temperature for 5 hrs. Aliquot samples from the donor and receiver sides were analyzed by LC/MS_MS, utilizing a generic LC methodology. Due to the excellent mass spectroscopic sensitivity, aliquots from 8 analytes were combined into one sample prior to analysis, thus increase throughput. This study demonstrates a convenient, sensitive and reproducible assay that allows rapid evaluation of passive drug permeability, thus provides very valuable information to predict oral bioavailability.