Getting the Most From Your Agilent HPLC Pump

Importance of the Topic

HPLC pumps are fundamental to liquid chromatography, directly influencing reproducibility, sensitivity and throughput. Proper pump operation and maintenance ensure reliable gradient formation, precise flow control and extended instrument lifetime, which are essential for routine analysis in research, industry and quality control labs.

Objectives and Study Overview

This application note guides users of Agilent Infinity series pumps to maximize performance through optimized system configuration, mobile phase management, scheduled maintenance and troubleshooting. It covers mixing schemes, dwell volume considerations, column selection, fluid path hygiene and best practice operating procedures.

Methodology and Used Instrumentation

Key topics and equipment:

• High-pressure vs. low-pressure mixing in binary, quaternary and flexible pump architectures
• Delay or dwell volume impact on retention time and resolution
• Agilent columns: Poroshell, ZORBAX SB-C18 and InfinityLab Performance Standard
• Mobile phase preparation: filtration, solvent bottles, inlet filters and Stay Safe Caps
• Degasser maintenance and compressibility calibration
• Purge, conditioning and prime cycles via manual valves or software-controlled functions
• Seal wash system with 90% water/10% isopropanol to prolong seal life
• Performance benchmarking using restriction capillary and InfinityLab LC Performance Standard
• Maintenance points: pump seals, frits, valves and peristaltic tubing
• Support resources: Agilent Community, YouTube, Lab Advisor and InfinityLab Assist Hub

Main Results and Discussion

Optimized low-dispersion UHPLC configurations achieved dwell volumes below 120 μL, leading to sharper peaks and consistent retention times. Regular mobile phase hygiene and filter replacement prevented cavitation and microbial growth, reducing baseline artifacts. Hydraulic performance monitored through ripple (<2%) and tuning values (–1 to +1) provided clear indicators of system health. Purging, conditioning and priming protocols effectively removed air and solvent residues, stabilizing pressure and flow before analysis.

Benefits and Practical Applications

Implementing these practices enhances data quality and method robustness across diverse applications including pharmaceutical QC, environmental screening and high-throughput metabolomics. Routine maintenance schedules reduce unplanned downtime and extend consumable life, while standardized benchmarking ensures compliance with regulatory and laboratory performance criteria.

Future Trends and Applications

Emerging trends include integration of predictive maintenance via cloud-based diagnostics, advanced pump designs for increased pressure and reduced dispersion, and automated solvent management systems. Adoption of greener solvents and sustainable chromatography workflows will drive further innovation in HPLC technology.

Conclusion

Adherence to recommended configuration, mobile phase management and preventive maintenance maximizes the lifetime and analytical performance of Agilent HPLC pumps. Leveraging available support tools and systematic benchmarking ensures consistent, high-quality chromatographic results.

References

Agilent Technologies application note DE-003661 'Getting the Most From Your Agilent HPLC Pump', January 14, 2025.