Ion Pair HILIC-MS: A Powerful Characterization Method for Biomacromolecule Analysis

The analysis of intact biomacromolecules is critical for understanding structure, modifications, and biological function—but conventional reversed-phase liquid chromatography (RPLC-MS) often falls short when it comes to separating closely related variants. In contrast, hydrophilic interaction chromatography (HILIC) offers complementary selectivity, making it a powerful alternative for resolving modifications and isomers that RPLC-MS cannot distinguish.

This presentation will focus on the development of HILIC for biomacromolecule characterization. It will highlight recent advances in ion-pair HILIC-MS and introduce a novel polymeric stationary phase for (IP)HILIC-MS. This poly(acrylamide-co-N,N-methylenebisacrylamide) monolithic stationary phase is designed to reduce unwanted secondary interactions and provides controlled morphology, which enhances separation capabilities compared to traditional silica-based materials.

Through real-world applications, including therapeutic oligonucleotides and intact monoclonal antibodies, learn how ion-pair HILIC-MS offers a powerful alternative to RPLC for biomacromolecule analysis. Attendees will see how this approach enables unprecedented resolution of impurities and glycoforms. Join us to explore how the potential of HILIC-MS technologies is pushing the boundaries of intact biomacromolecule characterization and opening new possibilities for biopharmaceutical analysis and quality control.

By attending this webinar, you will learn:

• Learn about the application and advantages of IP-HILIC-MS and acrylamide-based monoliths for the characterization of diverse biomacromolecules
• Discover how HILIC-MS technologies promise to enhance our understanding of complex biomolecules

Presenter: Andrea Gargano (Associate Professor of Analytical Chemistry, van't Hoff Institute for Molecular Science, University of Amsterdam)

Andrea Gargano is an Associate Professor of Analytical Chemistry at the van't Hoff Institute for Molecular Science, University of Amsterdam and part of the  Chemometrics & Advanced Separations Team (https://cast-amsterdam.org/). He received his Ph.D. in Analytical Chemistry from the University of Vienna and postdoctoral fellowship at the University of Amsterdam, with guest research across leading separation and mass spectrometry laboratories (e.g., PNNL, Northwestern University, LBNL).

The research of his team focuses on advancing high-resolution LC-MS technology, developing microscale and multidimensional separations for macromolecular analysis. He uses separation science to detect molecular structures that MS alone may miss. This includes hyphenated separations for biopharmaceutical and endogenous protein analysis, impurity profiling of oligonucleotides, and synthetic polymer characterization.