Agilent LC AND LC/MS - Columns Application Catalog

Importance of the Topic

Liquid chromatography remains a cornerstone of modern analytical chemistry, underpinning applications from pharmaceutical quality control to environmental monitoring. The ability to separate complex mixtures of small molecules, peptides, and biomolecules with speed, sensitivity, and reproducibility is essential for advancing drug discovery, process validation, food safety, and proteomics. Novel column technologies drive improvements in resolution, throughput, and method robustness, enabling laboratories to tackle new challenges and accelerate their workflows.

Objectives and Overview of the Document

This resource catalog provides a comprehensive guide to Agilent’s LC and LC/MS column portfolio, including conventional, high-performance, and ultrahigh-performance columns. It outlines key column technologies—porous-shell particles, sub-2 µm totally-porous packings, silica-embedded polar phases, polymeric packings, and specialized chemistries—and details selection strategies, method transfer guidelines, and practical considerations for column lifetime and compatibility.

Methodology and Instrumentation

The catalog covers columns compatible with a broad range of Agilent and third-party HPLC and UHPLC systems, including the 1200, 1260, and 1290 Infinity Series. It highlights column hardware options—Solvent Saver, narrow-bore, capillary, PrepHT, and cartridge formats—and recommends guard columns for lifetime extension. Guidance on tubing, fittings, and flow-cell configurations ensures minimal extra-column volume and optimal detector response for fast, high-efficiency separations.

Main Results and Discussion

Key technologies and their performance characteristics include:

• Poroshell 120: 2.7 µm superficially porous particles delivering sub-2 µm efficiency at 40–50% lower backpressure, across C18, C8, phenyl-hexyl, CN, Bonus-RP, and HILIC phases.
• ZORBAX Rapid Resolution (3.5 µm) and Rapid Resolution HT (1.8 µm) columns: fast, high-throughput separations with 600 bar stability; and RRHD (1.8 µm) columns for 1200 bar UHPLC applications.
• ZORBAX Eclipse Plus: double-endcapped C18 and C8 phases offering superior peak shape for acids, bases, and neutrals from pH 2–9 in 1.8, 3.5, and 5 µm formats.
• ZORBAX StableBond: sterically protected phases stable to pH 1–8 at elevated temperatures, including C18, C8, C3, phenyl, CN, and “Aq” for polar compounds.
• Polar-embedded chemistries: Bonus-RP (amide-embedded C18) and Extend-C18 (bidentate bonding) for improved peak shape of basic compounds and high-pH separations up to pH 11.5.
• HILIC Plus: non-bonded Eclipse Plus silica for retention of small polar analytes and LC/MS sensitivity in 1.8, 2.7, and 3.5 µm packings.
• Polymeric PLRP-S: styrene/divinylbenzene copolymer packings for pH 1–14 and macromolecule separations, including peptides, proteins, and nucleic acids (100–4000 Å pores).

Benefits and Practical Applications

The diverse column portfolio supports:

• Fast LC and high-resolution UHPLC method transfer: Poroshell 120 and RRHT/RRHD phases enable rapid adoption of sub-2 µm performance on both 400–600 bar and 1200 bar systems.
• Robust low-pH analysis: StableBond and Bonus-RP deliver stable, tail-free peaks for basic drugs and small molecules in TFA or formic acid mobile phases.
• High-pH selectivity: Extend-C18 expands the selectivity window by operating at pH 9–11.5 for free-base retention of amines and peptides.
• Polar compound retention: HILIC Plus and Polaris phases excel in separating highly polar metabolites, glycans, and small bioactive molecules without ion-pairing agents.
• Preparative scale-up: PrepLC, PrepHT, Pursuit, and Polaris Prep columns furnish scalable workflows from milligram to gram quantities with guard and cartridge systems for rapid exchange.

Future Trends and Potential Applications

Emerging directions include two-dimensional LC methods integrating orthogonal phase chemistries, further miniaturization into nanoLC for single-cell proteomics, and automation of method development through AI-driven column and mobile phase selection. Continued refinement of superficially porous particles and novel polymer hybrids will push speed and sensitivity limits, while expanded pH stability chemistries will unlock new separation spaces for challenging analytes.

Conclusion

Agilent’s comprehensive LC column portfolio addresses the full spectrum of analytical and preparative needs for small molecules and biomolecules. By combining innovative stationary phases, precise hardware, and robust method-transfer tools, users can achieve faster, more reproducible separations with minimal downtime and lower operating costs.