Highlights from the 54th International Symposium on High Performance Liquid Phase Separations and Related Techniques (HPLC 2025)

The 54^th International Symposium on High Performance Liquid Phase Separations (HPLC 2025), brought together 1111 scientists from 54 nations, representing a diverse and dynamic chromatographic community ready to share innovations, challenge conventions, and inspire the next generation of scientists (Figure 1). The conference was fully packed with separation science content. Automation and artificial intelligence/machine learning took center stage and is poised to shape the future of the laboratory. Oligonucleotides dominated the application spotlight, sustainability continued to increase its presence, and students and senior scientists alike found inspiration from posters, oral presentations, tutorial/workshops, interactions with vendors, and impromptu discussions.

In a similar fashion to the previous HPLC symposium review articles (1–7), many colleagues in attendance at the symposium were asked for their insights regarding the most interesting topics they observed at the event. What follows is a synopsis of these highlighted topics, along with some personal views and observations.

ChatHPLC?

The most talked-about topic from HPLC 2025 was artificial intelligence (AI) and its growing impact on separation science. Sessions on AI, machine learning (ML), and laboratory automation were well attended. While both enthusiasm and skepticism abound, Peter Schoenmakers offered a poignant viewpoint: “There is no ChatHPLC that will take over our jobs.” Workshops, short courses, and tutorials featuring AI and ML were conducted during the event, reflecting the growing interest in understanding how data-driven tools can (and cannot) aid method development. Of major concern to some is the quality of data AI/ML uses as a basis to resolve issues or make decisions. Although for many, AI is in its infancy regarding utility in separation science, AI and ML applications will likely continue to be a hot topic for years to come.

Oligonucleotides

As with the past few HPLC meetings (1,7), oligonucleotides and RNA therapeutics dominated the application space during HPLC 2025. Koen Sandra’s keynote set the tone for the conference, discussing structural analysis and reinforcing the need for orthogonal methods such as ion-pair reversed phase liquid (IP-RPLC), hydrophilic interaction liquid chromatography (HILIC), and anion-exchange (AEX) chromatography to more fully characterize these complex molecules. Multiple talks, including those by Dwight Stoll and Martin Gilar, emphasized pore design, column selectivity, and resolving power as key tools in dealing with and understanding these complex analytes.

Sustainability in Separation Science

Sustainability continues to grow in importance at the HPLC conference. As noted by Elia Psillakis, “[this was] the first time a session on sustainability appeared in the [HPLC] program.” Talks on greener solvents, reduced solvent usage through miniaturization, and method simplification garnered serious attention. Helena Hrušková’s award-winning poster on peptide hormone analysis in islet organoids was one example of environmental and analytical innovation aligning. Capillary LC and three-dimensional (3D)-printed columns were also touted for their role in solvent savings. Talks by Dave Anspach and Francesca Rigano suggested capillary scale might be the “sweet spot” between environmental benefits and performance.

Proteomics Advances

Proteomics, too, made a powerful appearance with Ryan Kelly, Lihua Zhang, John Yates, and Jennifer Van Eyk presenting visionary approaches from 5-minute single-cell workflows to new approaches to spatial proteomics. The excellent work in this field is providing new approaches to investigate diseases such as Alzheimer’s and cystic fibrosis.

Nurturing the Next Generation

One resounding theme was education. From Bob Pirok and Peter Schoenmakers’ new book (8) launch to a dynamic education session organized by Simone DiMartino, Andrea Gargano, and Martina Catani, the focus on teaching separation science was unmistakable. After a series of lightning talks on teaching innovation—ranging from newly published books and student modeling software to entrepreneurial learning and project-based curricula—the session culminated in a powerful outdoor group activity on the BMCC terrace. Participants co-designed the profile of the ideal analytical scientist of the future, identifying core knowledge (such as fundamentals, AI, and programming), key skills (for example, communication, critical thinking, and leadership), and essential mindsets (including motivation, empathy, curiosity, and openness).

Short courses covered topics from reversed-phase method design (Euerby) to (again) AI applications (Pirok and Bos), emphasizing the importance of both foundational and forward-thinking training.

One of the standout additions to this year’s HPLC Symposium was the Innovation Hub—a dedicated space led by Simone Dimartino and designed to spark creative thinking and collaborative problem-solving.

Together, these sessions enriched the technical program with a memorable, human-centered counterpoint—where creativity, clarity, and collective insight were as vital as chromatograms.

The early career scientists were not just learning, they were leading. Flash poster presentations gave these scientists a chance to shine, and attendees applauded their poise, creativity, and fundamental science. The careers event also drew significant interest, offering insights into industry trends and providing essential skill-building.

One of the most noted highlights of the conference was a presentation by one of our promising "next-generation" separation scientists, Gaëlle Spileers from Frederic Lynen's group at the University of Ghent. Spileers' study showcased the first successful integration of X-ray fluorescence spectroscopy (XRF) as a complementary, flow-through, universal detector for HPLC (Figure 2). XRF provides a molecule-independent, elemental-based response, much like ICP-MS, but without the sample preparation complexity or hyphenation headaches. While XRF is no stranger to analytical science, it has rarely been employed in dynamic separation work and never paired with HPLC before. This combination offers several perks: it quantifies compounds without the need for standards, accurately measures entire groups of compounds, and works happily alongside other detectors like MS to add elemental insights to structural elucidation. Although the sensitivity of this first-generation system is modest, it is not an inherent limitation of XRF; refinements in data collection and X-ray sources promise significant improvements. The result is proof-of-principle that HPLC–XRF can combine the separation power of chromatography with the universal, linear response of XRF, potentially offering the scientific community an affordable, robust, and refreshingly uncomplicated elemental detection option.

HILIC… I Must Mention HILIC

HILIC continues to emerge as a powerful and evolving platform owing to its growing role in the analysis of complex biomacromolecules and nucleic acid therapeutics. Advances in mechanistic understanding were highlighted by Mark Schure’s keynote and a poster from Hsiao-Feng Liu et al., where molecular simulations revealed direct solute interactions with silanols and ligands, challenging the conventional view of partitioning into a stagnant water layer. Application-driven talks demonstrated HILIC’s versatility: Jonathan Maurer used HILIC to assess lipid nanoparticle encapsulation efficiency in mRNA therapeutics, while Andrea Gargano’s group showcased ion-pairing HILIC for high-resolution impurity profiling of phosphorothioated oligonucleotides, overcoming the analytical challenge of 1 Da mass differences. Further innovation came from the development of acrylamide-based monolithic stationary phases for selective glycoform separations in intact monoclonal antibodies, achieving unprecedented resolution of subtle sugar-composition variants in 150 kDa proteins. Collectively, these contributions position modern HILIC, particularly when coupled with ion-pairing strategies, advanced stationary phases, and deeper mechanistic insight, as a critical tool for next-generation separations in therapeutic oligos, mRNA, and large biomolecules.

From Flash Presentations to Fundamental Philosophy

Poster presentations are a vital part of the HPLC conference. The poster sessions were vibrant and provided potential career springboards for early career as well as seasoned professionals. With 515 posters (247 in competition) and dedicated awards in areas like biopharmaceuticals, method development, and sustainability, the poster hall buzzed with energy. Helena Kim, Julius Schwieger, and Athanasios Tsalmpouris each walked away with major honors (Table I), and contributors like Xavier Subirats highlighted the importance of poster jury duty, noting “That special feeling you get in your gut when science comes alive.”

And So Much More…

Many, many more highlights were captured by attendees, including developments in multidimensional separations, slalom LC, and microfabricated stationary phases, to name a few. These talks and sessions were highlighted by speakers like Adam Woolley, Andrea Gargano, and Fabrice Gritti. In addition to the more academically centered scientific discussions, this year’s HPLC also boasted a specific industrial track. More details regarding the outcome of the industrial track initiative are promised in future LCGC International publications.

Conclusions

The 54th HPLC Symposium in Bruges offered a comprehensive and thoughtful view of the current state and future direction of liquid-based separation sciences. From emerging applications of AI and ML to advances in oligonucleotide analysis, sustainability-driven approaches, and continued progress in HILIC method development, the program reflected the diversity of ongoing work in the field. Presentations in proteomics, novel detection strategies, and mechanistic studies highlighted work to address complex analytical challenges. The emphasis on education and collaboration ensures that both established and early-career scientists are well equipped to contribute to future developments.

We now look ahead to HPLC 2026, to be held June 6–11, 2026, in Indianapolis, Indiana. This meeting promises to continue to provide a platform for sharing advancements, discussing challenges, and building the professional connections that support the progress of liquid phase separations worldwide.

Acknowledgements

Special thanks to the attendees whose thoughtful contributions shaped this review: Deirdre Cabooter, Elia Psillakis, Fabrice Gritti, Kevin Schug, Martina Catani, David McCalley, Jennifer Field, Xiaoli Wang, Paul Jacobs, Bob Pirok, Peter Schoenmakers, Frederic Lynen, Stefan Lamotte, Emanuela Gionfriddo, Tom Walter, Szabolcs Fekete, Steven Ray Wilson, Bo Zhang, Andrea Gargano, Frank Michel, Simone DiMartino, James Grinias, Xavier Subirats, Cary Simpson, and Egidijus Machtejevas.

And sincere congratulations to the organizing committee: Gert Desmet, Ken Broeckhoven, Deirdre Cabooter, Sebastiaan Eeltink, Frederic Lynen, and honorary chair Pat Sandra.

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