Comprehensive Metabolite Identification - Analysis of HepatoPac® Co-cultured Hepatocytes using SWATH™

The determination of metabolic stability and the accurate prediction of major human metabolites are critical for the optimization of drug candidates but traditional models for evaluating drug metabolism are short-lived and have a limited applicability for monitoring multi-generation metabolites or drug clearance kinetics. Furthermore, traditional LC/MS instrumentation approaches lack the sensitivity to measure metabolites within the therapeutic dosing range, often resulting in the need to generate metabolite profiling and drug clearance data from separate samples. This application note demonstrates the identification and quantitation of metabolites, as well as evaluated drug clearance kinetics from the same sample. Multi-generational drug metabolites obtained from a multi-species hepatocyte co-culture model (HepatoPac) were analyzed on a high-resolution TripleTOF® 6600 instrument with data-independent SWATH acquisition for simultaneous metabolite ID and relative quantitation across multiple time points.