Waters ACQUITY UPLC and ACQUITY PREMIER CSH Columns

Significance of the Topic

Efficient and reproducible chromatographic separations are critical across pharmaceutical, environmental, food-safety and biochemical laboratories. Waters ACQUITY UPLC and ACQUITY PREMIER CSH columns leverage Charged Surface Hybrid technology to improve peak shape and loading capacity for basic analytes under acidic, low-ionic strength conditions. The combination of sub-2 µm particles, robust surface chemistry and integrated quality control ensures high resolution, sensitivity and method reproducibility in demanding ultra-high performance liquid chromatography applications.

Objectives and Overview

This manual provides a full set of instructions and best practices for the care, installation, operation, cleaning, regeneration and storage of ACQUITY UPLC and ACQUITY PREMIER CSH columns. It also introduces eCord Intelligent Chip technology, which records individual column performance, manufacturing data and usage history to simplify quality assurance and lifecycle management.

Methodology and Instrumentation

Key procedural steps and operating guidelines include:

• Column installation: purging buffers, connecting inlet and outlet, installing VanGuard pre-columns or FIT cartridges, and verifying leak-free ferrule seating.
• Equilibration: flushing with ten column volumes of target mobile phase (including a pre-flush with water/organic mixture to avoid buffer precipitation) until stable backpressure is achieved.
• Operational limits: pH 1–11 (depending on chemistry), temperatures up to 80 °C (reduced at high pH), maximum pressure 18,000 psi, and use of HPLC-grade solvents filtered through 0.2 µm membranes.
• Cleaning and regeneration: tailored solvent sequences for polar, non-polar and proteinaceous contaminants (e.g., water, THF, methanol, DMSO or guanidine solutions), using 20 column volumes per step at elevated temperatures where safe.
• Storage: intermediate water flush followed by 100% acetonitrile for periods over 48 hours, sealed to prevent evaporation and salt precipitation.

Used Instrumentation

• Waters ACQUITY UPLC and ACQUITY Arc systems with column heater modules
• ACQUITY PREMIER CSH columns (C18, phenyl-hexyl and fluoro-phenyl chemistries)
• VanGuard pre-columns and Fully Integrated Technology (FIT) cartridges for inlet protection and easy replacement
• eCord Intelligent Chip for on-column data storage of batch certificates, test chromatograms and usage logs
• High-performance gradient pumps, detectors and ultra-pure water systems

Main Results and Discussion

Charged Surface Hybrid packing materials deliver high efficiency and peak symmetry for basic compounds, backed by narrow manufacturing specifications under cGMP and ISO 9001. Individual columns are performance-tested with acidic, neutral and basic probes, and results are stored on the eCord chip. Regular efficiency checks (theoretical plate counts and tailing factors) help track column aging and ensure method consistency.

Benefits and Practical Applications

The combination of CSH stationary phases, guard devices and eCord tracking offers:

• Extended column lifetimes through targeted cleaning, regeneration and FIT cartridge replacement
• Reliable QC documentation with per-column certificates and performance test chromatograms
• Simplified method transfer and troubleshooting via recorded usage history
• Enhanced analyte recovery and sensitivity using MaxPeak high-performance surfaces

Future Trends and Opportunities

Advances may include deeper integration of smart chip technologies with laboratory information management systems (LIMS), AI-driven column maintenance alerts, novel surface chemistries for extreme pH or chiral separations, greener solvent systems and expanded miniaturized formats for high-throughput applications.

Conclusion

Adhering to recommended installation, operation, cleaning and storage protocols maximizes the performance and lifetime of ACQUITY UPLC and ACQUITY PREMIER CSH columns. The eCord Intelligent Chip further streamlines quality assurance by preserving complete manufacturing and usage data, ensuring confidence in every chromatographic analysis.

Reference

• Waters Corporation. Care and Use Manual for ACQUITY UPLC and ACQUITY PREMIER CSH Columns. October 2020.