GLP-1 peptide drugs such as semaglutide and tirzepatide are transforming treatment for diabetes and weight management. As these compounds move from innovators to generics, chromatographers face mounting pressure to deliver reproducible, high-resolution methods for complex peptide mixtures.
In a new episode of Concentrating on Chromatography, produced in collaboration with Separation Science, Phenomenex’s Principal Marketing Development Manager, Sean Orlowicz, detailed the analytical challenges driving advances in peptide separations and the company’s approach to supporting laboratories across the pharmaceutical pipeline.
Peptide Complexity and Analytical Demands
Unlike small molecules, peptide drugs sit in a gray zone between traditional pharmaceuticals and biologics. Their impurities often differ by a single amino acid deletion, oxidation, or isomerization—minute variations that are difficult to resolve chromatographically. Orlowicz emphasized that these subtle differences require exceptional selectivity and efficiency to separate related substances accurately.
Phenomenex’s ARIS Peptide XB-C18 column exemplifies this focus. Built on a core-shell particle for enhanced efficiency, it offers the selectivity needed to distinguish closely related peptide species while maintaining high resolution and reproducibility. Orlowicz explained that this chemistry has become a mainstay for both analytical and preparative workflows, from early-stage R&D to large-scale manufacturing.
Method Robustness in a Global Regulatory Landscape
As GLP-1 therapeutics reach massive patient populations, regulatory scrutiny intensifies. These injectables demand validated, transferable methods that can withstand variations across global contract labs and manufacturing partners.
Orlowicz compared method transfer to sharing a recipe—success depends not only on ingredients but also on execution. Robustness testing across multiple instruments, column batches, and operators is essential to ensure consistency in impurity profiling and stability testing.
The Expanding Role of High-Resolution Mass Spectrometry
High-resolution mass spectrometry (HRMS) is emerging as a key complement to chromatographic separation. Beyond research applications, HRMS offers process analytical and quality-control benefits by distinguishing subtle mass differences that can elude LC-only workflows. Orlowicz noted growing adoption of MS in regions where the technique was once cost-prohibitive, as labs recognize its potential to overcome challenges posed by shallow gradients and structurally similar impurities.
Common Pitfalls and Best Practices
For analysts entering peptide workflows, Orlowicz offered practical guidance:
- Control temperature—both mobile phase and column—to improve method robustness.
- Leverage vendor expertise and published applications for troubleshooting and optimization.
- Be patient—peptide separations often require longer gradients and higher pressures than small-molecule assays.
- Avoid “copy-paste” method design—customize gradients, ion-pairing agents, and stationary phases to the peptide’s unique chemistry.
He also underscored the value of size-exclusion chromatography for aggregation studies, which ensures peptide stability and potency in sterile injectable formulations.
Commitment to Education and Innovation
Phenomenex continues to expand its peptide analysis portfolio through technical publications, webinars, and application notes that feature real-world chromatograms, not just standards. Recent studies explored separations for semaglutide and tirzepatide impurities, while new size-exclusion columns target smaller, hydrophobic peptides typical of GLP-1 analogs.
The company’s strategy extends beyond peptides to oligonucleotides and AAVs, reflecting its broader mission to develop chromatographic tools for the next generation of complex therapeutics.
Outlook: Precision Peptide Analysis for the Decade Ahead
As GLP-1 drugs continue reshaping global healthcare, advances in LC and MS will be central to ensuring their safety, efficacy, and accessibility. Orlowicz predicts sustained innovation in preparative chromatography, with manufacturers striving for faster, more economical purification processes without sacrificing purity.
“The enemy of innovation,” he remarks, “is the existence of copy and paste.” In the evolving world of peptide therapeutics, fresh thinking and robust chromatographic design will define the leaders of tomorrow’s analytical science.
Explore Additional Resources
Learn More:
- Visit Phenomenex.com for GLP-1 application notes and technical support
- Explore the Concentrating On Chromatography podcast to dive into the frontiers of chromatography, mass spectrometry, and sample preparation with host David Oliva.
- Download the complete Phenomenex technical note on semaglutide impurity separation.
Connect with Sean:
- LinkedIn: Sean Orlowicz
