At Analytica USA 2025, Martin Gilar, a distinguished Research Fellow within the Separations R&D Group at Waters Corporation, took center stage in Columbus, Ohio. His captivating presentation, “RNA Analysis with LC and Affinity LC Methods,” delved into the realm of mass spectrometry as a potent tool for dissecting various RNA species. The complexity of long RNA molecules poses a challenge for comprehensive analysis, leading to the innovative approach of fragmenting them into more manageable pieces for sequence reconstruction. Gilar unveiled a groundbreaking method harnessing novel RNase enzymes to generate fragments of optimal length tailored for LC-MS analysis. Moreover, he addressed the intricacies of denaturing and non-denaturing conditions, offering insights into measuring the RNA melting temperature (Tm) to facilitate the selection of suitable analytical conditions for diverse RNA molecules.
Embarking on his scientific journey at the Institute of Chemical Technology in Prague, Gilar obtained his Ph.D. in analytical chemistry in 1996. Subsequently, his postdoctoral endeavors in Boston were dedicated to pioneering separation techniques for oligonucleotides. Since 1998, Gilar has been a pivotal figure at Waters Corporation, spearheading research initiatives encompassing columns, sample preparation, and cutting-edge instrumentation. With a profound expertise in biopolymers and nucleic acids, he has authored a remarkable 75 papers elucidating these domains throughout his illustrious career.
A recent collaboration between Gilar and the esteemed LC Troubleshooting columnist, Dwight Stoll, culminated in a seminal paper titled, “Challenges and Solutions in Oligonucleotide Analysis, Part I: An Overview of Liquid Chromatography Methods and Applications.” In this collaborative effort, Gilar sheds light on how innovative RNase enzymes can revolutionize RNA mapping via LC–MS in comparison to traditional enzymes. He delves into strategic approaches to mitigate ambiguous digest products, highlights the primary constraints in reconstructing lengthy RNA sequences from enzymatic digests using LC–MS, and expounds on how his methodology adeptly addresses issues such as overlapping fragments or incomplete digestion. Emphasizing the pivotal role of melting temperature in LC analysis of nucleic acids, Gilar’s insights provide a comprehensive framework for navigating the complexities of nucleic acid analysis.
As an integral segment of our dialogue with Gilar, this interview snippet offers a glimpse into the wealth of knowledge shared at Analytica USA. Stay tuned as we unravel the tapestry of insights gleaned from this prestigious conference in the forthcoming coverage. For real-time updates on the latest revelations from Analytica USA, remember to click here.
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In conclusion, the realm of liquid chromatography analysis of nucleic acids represents a captivating intersection of innovation and precision. Gilar’s pioneering work not only underscores the significance of leveraging advanced methodologies but also highlights the intricate nuances involved in unraveling the mysteries of RNA analysis. As we navigate the ever-evolving landscape of analytical sciences, the insights shared by Gilar serve as a beacon illuminating the path towards enhanced understanding and proficiency in nucleic acid analysis. Stay tuned as we continue to explore the frontiers of chromatography and molecular analysis, delving deeper into the realms of science and technology to unravel the complexities of nucleic acid analysis.
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