SFC 2025 Europe Preview: The Role of SFC in Life Science Analysis

What specific advantages does modern supercritical fluid chromatography (SFC) offer over traditional high performance liquid chromatography (HPLC) when analyzing chiral compounds in pharmaceutical development?

John Reily: The integration of supercritical fluid chromatography (SFC) for chiral separation has allowed for productivity increases, decreases in toxic/hazardous solvent consumption, and a greener chromatographic approach. Ultimately, this leads to a decrease in time for the “drug-design-test” cycle for drug discovery projects.
Maria Kristen Parr: Particularly for preparative applications, SFC methods allow for easy product recovery. The analytical-scale method development helps to later upscale methods without a huge amount of effort. In addition, in the analytical scale, the decrease in solvent use and waste generation is considered beneficial. It has also been demonstrated that SFC may resolve chiral drugs and chiral impurities or metabolites in one single run, where HPLC may sometimes struggle.

How is SFC currently being applied in biopharmaceutical workflows, such as the analysis of peptides, oligonucleotides, or biologics?

JR: At Novartis, we are currently investigating where SFC “orthogonality” can provide benefits for more complex modalities. We have demonstrated that for some cyclic peptides, SFC is beneficial, although we need to invest more time in this space to have more definition on the prediction of when SFC may be of more value here than traditional reversed-phase (RP) HPLC approaches.
MKP: Our data on the separation of antibiotic peptides clearly demonstrate that SFC is a useful tool in peptide mixture separation. Also, in the case of insulins, we have demonstrated that SFC is a very useful technique. Extending the approach to larger peptides and proteins is worth exploring, particularly when using high modifier levels that enable gradients spanning both classical SFC and HPLC conditions within a single run—covering amino acids, small and large peptides, and even intact proteins, for improved impurity profiling. As mentioned above, I foresee SFC as also being very useful for chiral separations in oligo analysis.
LM: SFC is being explored for more molecules beyond small molecules, including peptides and oligonucleotides. While some interesting work has been published on SFC for peptides and oligonucleotides, because of the high polarity of some of these molecules, it is far from a universal technique for the analysis of these types of molecules. LC is still the go-to technique for the analysis and purification of most biotherapeutic molecules.

In drug discovery pipelines, how does SFC contribute to high-throughput purification and screening of lead compounds?

JR: It has been fundamentally demonstrated throughout the industry that within drug discovery, SFC speeds up the purification step, which ultimately will lead to more efficient screening of lead compounds.

LM: Most purification laboratories in pharmaceutical research are using SFC to increase productivity and reduce cycle time for chiral and achiral high-throughput workflows. The use of SFC reduces purification times, solvent usage, and waste generation, and results in lower purification costs.

Can SFC be effectively coupled with mass spectrometry (SFC-MS) for metabolomics or lipidomics applications in biomedical research?
JR: There have been a range of publications to demonstrate SFC-MS applicability for bioanalysis applications within drug discovery; however, I still believe that this opportunity has not been taken up as much because RP LC–MS has such a broad applicability here.
MKP: The high chromatographic separation capability of SFC may ease proper compound identification, especially in cases where large numbers of potential isomers are expected. In that sense, lipidomics, lipid profiling, and characterization of nanoparticle composition may be fields of interest in the future.
LM: Coupling MS to SFC is very easy and has been used for almost 30 years. Significant research has been published over the past 15 years on the use of SFC-MS in the lipidomics and metabolomic fields.

Biographies
John Reilly is an associate director in the Department of Global Discovery Chemistry at Novartis Institute of Biomedical Sciences, and is based in Basel, Switzerland. He currently leads the separations team, where there is a focus on providing purification and analytical characterization methodologies supporting a chemistry unit with over 250 chemists. An advocate of knowledge exchange, Reilly has had global roles in discovery and development during his time at Novartis and previously with Eli Lilly. He completed his MSc. At Birkbeck College in 1992 and his PhD in analytical science at Imperial College in 2002 after his initial BSc in chemistry at Reading University, Reading, UK in 1989. His research goals include the promotion of SFC’s new chromatographic characterization and purification methods and physicochemical affinity screens to investigate drug-like properties of molecules. Reilly has published more than 50 articles and has been a board member of The Chromatography Society. He has also been on the editorial board of Chromatography Today and American Journal of Modern Chromatography.

Larry Miller is an Associate Director in the Discovery Analytical group at Novartis in Cambridge, MA. He has spent his career performing small molecule achiral and chiral purifications at the microgram to multi-kg scale. At Novartis, Larry manages a group responsible for analytical and purification support to the Global Discovery Chemistry function. Previously, he spent 20 years at Searle/Pharmacia, and 20 years at Amgen. Larry has over 50 peer-reviewed publications and over 55 presentations at scientific meetings and he has served as co-instructor for SFC short courses in the US, Europe, and Asia. He has been involved with the Green Chemistry Group since its inception in 2007 and currently serves as President of the group. He earned his master’s in chemistry from Roosevelt University and his bachelor’s in chemistry from the University of Illinois at Urbana-Champaign, USA.

Maria Kristina Parr has worked as a professor for pharmaceutical analysis at Freie Universität Berlin, Germany, since 2012. The research focus of her group is mass spectrometric analysis hyphenated to different chromatographic separation techniques (GC–, LC–, and SFC-MS[/MS]), with a main focus on drug, supplement, and bioanalysis as well as anti-doping research. Investigations in drug metabolism, determination of endogenous and xenobiotic compounds, and drug-drug interactions play an important role in her research. Research activities in her team consider green and white analytical chemistry principles, quality management, and analytical quality-by-design.