Perform Rugged, Fast LC with Confidence - Agilent InfinityLab Poroshell 120 columns for HPLC and UHPLC

Importance of the Topic

Superficially porous particle technology bridges the gap between traditional HPLC and modern UHPLC by offering high efficiency at moderate pressures. Agilent InfinityLab Poroshell 120 columns leverage a unique single-step shell process to deliver robust reproducibility, extended lifetime, and broad chemical stability. This innovation supports faster separations, reliable method transfer, and high throughput across diverse applications in pharmaceuticals, environmental testing, food safety, and forensic analysis.

Objectives and Study Overview

This application note introduces the Agilent InfinityLab Poroshell 120 column family, detailing its design, available particle sizes (1.9 µm, 2.7 µm, 4 µm), and eighteen bonded phases. It aims to demonstrate performance advantages—efficiency, speed, reproducibility—and practical benefits such as simplified method development, seamless HPLC/UHPLC transfer, and compatibility with LC/MS workflows.

Methodology and Instrumentation

• Manufacturing: A three-step process forms a solid silica core, applies a uniform porous shell, and bonds stationary-phase chemistries to ensure narrow particle size distribution.
• Particle Family: Choice of 1.9 µm for maximum UHPLC efficiency, 2.7 µm for UHPLC at lower pressures, and 4 µm for enhanced HPLC performance.
• Chemistries: Includes EC-C18, SB-C18, EC-C8, SB-C8, HPH-C18/C8, HILIC variants (HILIC, HILIC-Z, HILIC-OH5), Bonus-RP, PFP, phenyl-hexyl, EC-CN, and four chiral phases (CD, CF, V, T).
• Instrumentation: Demonstrated on Agilent InfinityLab 1220, 1260, and 1290 LC/UHPLC systems, with LC/MS validation using Agilent MSD iQ and 6410 QQQ.
• Operating Conditions: Pressure ratings up to 1300 bar (for 1.9 µm), temperature stability to 90 °C, and pH compatibility from 1 to 12 depending on phase.

Main Results and Discussion

• Reproducibility: Five lots of 2.7 µm EC-C18 columns produced overlapping chromatograms over 2.5 years, confirming outstanding lot-to-lot consistency.
• Efficiency and Speed: The 1.9 µm column achieved baseline resolution of six analytes in 0.35 min at 1150 bar. The 2.7 µm column matched >90% of sub-2 µm performance at half the pressure.
• Method Transfer: Legacy USP naproxen methods ran 4.5× faster on a 4 µm Poroshell column without changing flow rate or mobile phase. A 2.7 µm Poroshell column halved run time and doubled plate count compared to the original 5 µm column.
• pH-Range Robustness: SB-phases delivered stable performance at pH 1–6, while hybrid HPH-phases maintained performance after 2000 injections at pH 10.
• Alternative Selectivity: PFP, phenyl-hexyl, Bonus-RP, EC-CN, and SB-Aq exhibited orthogonal selectivity to C18, improving separation of isomers, vitamins, catechins, and explosive residues.
• Chiral Separations: Superficially porous chiral columns resolved amphetamine enantiomers in under 5 min versus ~15 min on fully porous analogs.
• HILIC Performance: HILIC-Z and HILIC-OH5 phases enabled MS-compatible separation of amino acids and B vitamins without ion-pair reagents.
• Column Lifetime: 2.7 µm and 4 µm columns with reinforced inlet frits resisted clogging, sustaining thousands of injections with minimal performance loss.

Benefits and Practical Applications

• Accelerated Throughput: Run times reduced up to 5× while maintaining resolution, boosting daily sample capacity.
• Rapid Method Development: Eighteen chemistries and scalable particles simplify selectivity screening and method transfer across instruments.
• Cost Efficiency: Lower operating pressures and extended column life reduce instrument wear, solvent use, and downtime.
• Versatility: Columns cater to reversed-phase, HILIC, polar ionic, and chiral modes, addressing needs in pharma, food safety, environmental, and forensic labs.
• Enhanced LC/MS Sensitivity: MS-friendly mobile phases and high-pH stability improve ionization of basic analytes.

Future Trends and Applications

Upcoming developments include expanded superficially porous phases tailored for large biomolecules and peptides, integrated digital ID tags for automated column tracking, and deeper integration with next-generation LC/MS platforms. Enhanced HILIC and zwitterionic phases will meet rising demand for metabolite profiling and polar compound analysis. Continued miniaturization and multiplexed column formats promise further speed and throughput gains.

Conclusion

Agilent InfinityLab Poroshell 120 columns combine superior manufacturing reproducibility, robust chemical stability, and a broad chemistry portfolio to elevate both HPLC and UHPLC performance. Their scalable particle sizes and diverse bonded phases enable faster, more efficient, and highly reproducible separations, empowering laboratories to improve throughput, reduce costs, and address complex analytical challenges.