Exploring LC Analysis of Nucleic Acids

At Analytica USA in Columbus, Ohio, Martin Gilar, who is a Research Fellow in the Separations R&D Group at Waters Corporation, delivered a talk titled, “RNA Analysis with LC and Affinity LC Methods” (1). Gilar’s talk focused on mass spectrometry, and how it is a powerful tool for analyzing various RNA types. Long RNA molecules are difficult to analyze as a whole, so they can be digested into smaller fragments and their sequence reconstructed (1). Gilar’s talk addressed a new method using novel RNase enzymes, which produces optimal-length fragments for LC-MS analysis (1). Additionally, the talk addresses challenges with denaturing and non-denaturing conditions, presenting a method for measuring the RNA melting temperature (Tm) to aid in selecting appropriate analytical conditions for different RNA molecules (1).

Gilar got his start at the Institute of Chemical Technology in Prague, where he earned his Ph.D. in analytical chemistry in 1996 (1). After receiving his Ph.D., he conducted postdoctoral research in Boston, which was focused on developing separation methods for oligonucleotides. Since 1998, he has worked at Waters Corporation, contributing to research on columns, sample preparation, and instruments (1). Gilar’s specialization is in the analysis of biopolymers and nucleic acids, publishing 75 papers on the subject over his career (1).

Recently, Gilar collaborated with LC Troubleshooting columnist and co-host of Analytically Speaking Dwight Stoll on a paper titled, “Challenges and Solutions in Oligonucleotide Analysis, Part I: An Overview of Liquid Chromatography Methods and Applications” (2). In this interview segment, Gilar addresses how novel RNase enzymes can improve RNA mapping by LC–MS compared to conventional enzymes, the strategies that can minimize ambiguous digest products, the main limitations in reconstructing long RNA sequences from enzymatic digests using LC–MS, and how his approach addresses issues such as overlapping fragments or incomplete digestion, and the importance of melting temperature for LC analysis of nucleic acids.

This interview segment is part of our conversation with Gilar. Throughout the next week, we will be publishing our coverage of the Analytica USA conference. To stay up to date with all the latest from our coverage of Analytica USA, click here.

References

1. Analytica USA, Martin Gilar. Analytica USA. Available at: https://aus2025.mapyourshow.com/8_0/sessions/speaker-details.cfm?speakerid=8 (accessed 2025-09-10).
2. Gilar, M.; Stoll, D. R. Challenges and Solutions in Oligonucleotide Analysis, Part I: An Overview of Liquid Chromatography Methods and Applications. LCGC Int. 2025, 2 (7). Available at: https://www.chromatographyonline.com/view/oligonucleotide-analysis-chromatography-methods-therapeutic-nucleic-acids