Denaturing SEC-MS Analysis Of High Molecular Weight Impurities In The GLP-1a Lipopeptides Semaglutide And Tirzepatide

By Stephan M. Koza, Philip Wuthrich, Stephen J. Shiner, and Matthew A. Lauber

Understanding high–molecular weight species (HMWS) is essential for ensuring the quality and stability of GLP‑1–based therapeutics like semaglutide and tirzepatide. Explore how denaturing size‑exclusion chromatography coupled with high‑resolution mass spectrometry (dSEC‑HRMS) enables detailed characterization of low‑abundance oligomeric impurities, even when they remain non‑dissociable under harsh conditions. The method provides accurate monomer mass assignments, clear differentiation of dimer and trimer forms, and insight into truncated variants that may arise during storage or degradation. Paired UV detection at 280 nm supports reliable quantitation of both HMWS and monomer concentration. For teams working on peptide stability, formulation, or analytical development, learn more about a blueprint for deeper impurity profiling and analytical considerations needed to interpret complex lipopeptide behavior.

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