Surviving Chromatography - Part 1: Preventive Action

Importance of the Topic

High-performance liquid chromatography (HPLC) and LC-MS are essential tools in analytical chemistry, but system downtime and poor reproducibility can undermine results and productivity. Preventive maintenance, use of high-quality consumables, proper sample preparation and method development are critical to sustain reliable performance, reduce troubleshooting time and lower total cost of ownership.

Objectives and Study Overview

This application note examines preventive actions to ‘‘survive’’ chromatography by addressing critical instrument supplies, sample-handling practices and method development strategies. The goals are to identify common sources of performance loss, compare original and third-party consumables, and recommend maintenance schedules to ensure stable retention times, peak shapes and detector sensitivity.

Methodology and Instrumentation

Key elements of the preventive approach include:

• Instrument supplies: Original Agilent solvent inlet filters, PTFE frits, pump piston seals, check valves, rotor seals and autosampler parts.
• Detector care: Maintenance of UV lamps, flow cells and proper tubing to minimize extra-column volume.
• Sample-cleanup tools: Filtration (syringe filters, in-line filters), solid-phase extraction cartridges/96-well plates, QuEChERS kits and Agilent Captiva EMR-Lipid sorbent to remove proteins and lipids.
• Column portfolio: Selection of InfinityLab Poroshell superficially porous phases (C18, SB-C18, HPH-C18, phenyl-hexyl, HILIC) and use of Quick Connect/Quick Turn fittings to reduce dead volume.

Key Results and Discussion

• Solvent inlet filters from Agilent exhibit uniform pore size and high cleanliness, eliminating contamination peaks observed with third-party filters.
• PTFE frits and pump seals of defined particle size and polymer blends extend lifetime, reduce pressure ripple and prevent leaks or pump blockages.
• Redesigned rotor seals and check valves deliver consistent sealing over tens of thousands of cycles and lower pressure fluctuations.
• Agilent deuterium lamps achieve up to 60 % higher UV intensity at low wavelengths, yielding improved signal-to-noise ratios and lamp life beyond 2000 hours.
• Sample cleanup with Captiva EMR-Lipid dramatically lowers lipid-related ion suppression, improves reproducibility (RSD < 3 %) and boosts analyte signal compared to protein precipitation alone.
• Bio-inert PEEK-lined HILIC-Z columns maintain sharp peak shapes for phosphorylated metabolites by preventing metal-analyte interactions, aided by regular wash protocols.

Benefits and Practical Applications

Adopting these preventive measures offers:

• Greater instrument uptime and fewer unscheduled repairs.
• Improved chromatographic reproducibility and sensitivity.
• Lower long-term maintenance costs and extended column lifetimes.
• Streamlined method development with a broad portfolio of stationary phases and easy-to-use fittings.

Future Trends and Applications

Emerging directions include automated, in-line sample cleanup integrated with chromatography, advanced stationary phases for wider pH and temperature ranges, artificial-intelligence-driven diagnostics to predict maintenance needs, and further miniaturization of LC systems to accelerate analyses while conserving solvents.

Conclusion

Preventive maintenance, original high-quality consumables, vigilant sample preparation and systematic method development are the cornerstones of robust chromatography. By planning ahead and adopting best practices, analytical laboratories can maximize performance, reduce downtime and achieve consistent, accurate results.

References

Powell M., Ucci A. Surviving Chromatography Part 1: Preventive Action. Agilent Technologies, September 2018.