Agilent InfinityLab preparative LC columns

Significance of the Topic

Preparative liquid chromatography is a cornerstone technique for isolating and purifying small molecules, peptides, and proteins in research, manufacturing, and quality-control laboratories. Optimizing column performance in terms of purity, yield, and throughput directly impacts operational costs, sample integrity, and time-to-result. Advanced stationary phases and workflows are essential to meet diverse purification challenges and support seamless scale-up from analytical screening to bulk production.

Study Objectives and Overview

This article introduces the Agilent InfinityLab family of preparative HPLC columns and associated purification solutions. The main goals are to present how these columns:

• Deliver robust, long-lived performance
• Enable predictable scale-up from analytical to preparative scale
• Support tailored workflows for high-throughput or high-yield objectives

By showcasing key data and method examples, the study demonstrates strategies to maximize purity, throughput, and yield across a range of applications.

Methodology and Instrumentation

Preparative separations were conducted on Agilent Infinity II LC purification systems, employing both analytical-scale (1220, 1260, 1290 Series) and preparative-scale (1260, 1290 Series) configurations. Stationary phases evaluated include:

• InfinityLab Poroshell 120 (superficially porous C18) for high-speed workflows
• InfinityLab Pursuit XRs (high surface area C18, C8, Diphenyl) for maximum loadability
• InfinityLab ZORBAX phases (Eclipse Plus C18/C8, Phenyl-Hexyl) for broad selectivity options

Mobile phases, flow rates, and injection volumes were adjusted to illustrate method transfer from sub-2 µm UHPLC columns to 4–5 µm preparative columns without loss of resolution.

Main Results and Discussion

Lifetime studies on a C18 preparative column under high flow conditions demonstrated stable peak shapes over >1000 injections. Scale-up experiments showed comparable retention and resolution when transferring a gradient method from a 1.8 µm analytical column to a 5 µm preparative column with increased inner diameter and flow rate. Application to curcuminoid purification highlighted two workflows:

• High-throughput mode on Poroshell 120 enabling 30 mg separation in under 3 minutes
• High-yield mode on Pursuit XRs delivering 120 mg per injection with clear resolution of bisdemethoxycurcumin, demethoxycurcumin, and curcumin

These results validate the columns’ robustness, scalability, and adaptability to diverse purification targets.

Benefits and Practical Applications

Agilent InfinityLab preparative columns offer:

• Long lifetimes and predictable performance, reducing rework and cost-per-sample
• Seamless method scale-up from analytical screening to bulk purification
• A diverse portfolio of chemistries and particle technologies to optimize for throughput or yield
• Compatibility with a unified LC platform for easy integration and maintenance

These advantages support workflows in pharmaceutical development, natural product isolation, and industrial process validation.

Future Trends and Opportunities

Emerging trends in preparative chromatography include:

• Further development of superficially porous and hybrid particle technologies to improve efficiency and load capacity
• Integration of real-time analytics and process control software for automated fraction collection and purity assessment
• Expansion of preparative solutions for biomolecules, such as monoclonal antibodies and oligonucleotides, leveraging larger pore media and tailored chemistries
• Adoption of greener solvents and reduced solvent consumption through miniaturized preparative formats

These innovations will drive higher productivity and sustainability in purification workflows.

Conclusion

Agilent InfinityLab preparative LC columns and purification solutions provide a versatile and reliable platform for optimizing purity, throughput, and yield across a wide range of applications. Their robust design, predictable scale-up, and extensive chemistry choices enable laboratories to address evolving purification demands efficiently and cost-effectively.