Metabolite Identification With HµREL Micro Livers

Poster on improving metabolite identification and metabolic stability assessment using predictive in vitro testing with HμREL® technology.

Poster Authors:

Sam Morris, Louise Telford, Adam Hughes, Katherine Fenner

Pharmaron UK Ltd, Hertford Road Hoddesdon Hertfordshire, EN11 9FH, UK

Overview of Metabolic Identification for Drug Discovery

Early-phase drug development hinges on understanding how compounds are metabolized. One of the most critical tools in this process is metabolite identification (Met ID)—the analysis of how a new chemical entity (NCE) transforms within biological systems. Using HµREL® micro livers, researchers at Pharmaron have developed a high-throughput, predictive assay that reveals key insights into metabolic stability and clearance, even for low-turnover compounds.

The Advantage of HµREL Micro Livers in Metabolic Stability Testing

Extended Incubation for Low Clearance Compounds

Traditional systems using suspended hepatocytes face a time barrier—cells remain viable for only around four hours, limiting the detection of slow-forming metabolites. HµREL® micro livers overcome this by maintaining cell function for up to five days, allowing for deeper metabolic profiling of slow-clearance drugs.

Key Benefits:

Increased sensitivity: Identifies metabolites not visible in traditional systems.
Greater efficiency: One sample can support both clearance and Met ID testing.
Scalability: 384-well plate format supports high-throughput screening.

Capabilities

Knowledge Center

Metabolite Profiling Using AMS

Metabolite Identification

Metabolism

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A Look at the Discovery Metabolite Identification Process

Integrated Workflow Using HµREL®

The workflow begins with incubation in HµREL® plates, using test compounds at 1 µM. Samples are collected at multiple time points (4, 24, 48, 72 hours) and processed with Sciex QTOF-MS systems. Using both CID and EAD fragmentation techniques, metabolites are structurally elucidated with precision.

Example: Warfarin, a slow-clearance compound, produced no metabolites at 4 hours but revealed three known metabolites at 72 hours, consistent with the literature.

Validated for Accuracy

The assay was validated using ten compounds across a range of clearance profiles. Results were consistent with in vivo data, establishing the system’s reliability in predicting intrinsic clearance (CLint) and supporting decision-making in early discovery.

Why This Matters in Modern Drug Discovery

The ability to conduct fast, predictive, and cost-effective Met ID testing allows researchers to:

Reduce development timelines
Lower compound usage
Identify metabolite liabilities earlier

By combining HµREL®’s micro liver technology with high-resolution mass spectrometry and tailored analytics, Pharmaron’s platform provides unmatched clarity for preclinical research teams.

References:

Download the full poster for a detailed walkthrough of the Met ID method using HµREL® and real data examples like Warfarin. Learn how your pipeline could benefit from more predictive in vitro testing.