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Day 2: TP28
Immobilized Artificial Membrane (IAM) Chromatography with Mass Spectrometric Detection: a Rapid Method for Screening Drug-Membrane Interactions

Hanlan Liu, Guy T. Carter, and Mark Tischler
Discovery Analytical Chemistry, Wyeth-Ayerst Research, 401 North Middletown Road, Pearl River, New York 10965

Immobilized artificial membranes (IAMs) are solid membrane mimetics that contain cell membrane phospholipids. They are covalently bonded to the surface of a silica chromatographic support to generate a phospholipid monolayer. IAM chromatography has been used in the measurement of drug-membrane partitioning, in predicting bile salt-membrane interactions, and in drug transcellular absorption studies. In those studies, the assay was conducted utilizing HPLC with UV detection where one compound is analyzed in a single experiment under isocratic conditions. In order to analyze a larger group of diverse compounds using an automated HPLC-UV system, a relatively long run time has to be used to ensure full elution off the IAM column. This approach clearly limits the throughput of the assay. Here we report a high-performance liquid chromatography (HPLC)/electrospray mass spectrometry method for measuring drug-membrane interaction using immobilized artificial membrane (IAM) fast-screening mini-columns. The HPLC mobile phase consists of phosphate buffered saline (i.e., 5.0 mM phosphate buffer at pH 7.4, 1.35 mM KCl, and 68.5 mM NaCl) and acetonitrile. This method facilitates the measurement of the IAM retention times of over ten compounds in one experiment, significantly reducing the analysis time as compared to the earlier IAM HPLC method. The particular electrospray source used demonstrates the ability to tolerate the high salt-containing nonvolatile buffer used for retention time measurement.

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