Metabolic Stability Assay Using Human Hepatocyte Co-cultures and Integrated Qualitative/Quantitative High Resolution Mass Spectrometry

Traditional metabolic stability methodology using suspended hepatocyte for drug candidates screening is limited in their ability to accurately predict clinical outcomes. Hepatocyte co-culture platform is a bioengineered, in vitro system with a defined cyto-architecture that provides sustained hepatic functions for at least four weeks1. In this presentation, we investigated human hepatocyte co-cultures model using an integrated qualitative/quantitative high resolution mass spectrometry approach to assess metabolic stability using TOF-MS and Sequential Window Acquisition of All THeoretical fragment ion spectra (SWATH) for non-targeted metabolite MS/MS analysis. The assay was evaluated using propranolol, lorazepam, ranitidine, and zoniporide, at clinically relevant concentration (1 μM) in 96-well format.