Agilent ZORBAX Rapid Resolution High Definition SB-C18 Threaded Column

Significance of the Topic

In ultra-high-performance liquid chromatography (UHPLC), achieving rapid separations with high resolution is vital for pharmaceutical, environmental, and industrial laboratories. Columns that maintain efficiency under extreme pressures, wide pH ranges, and elevated temperatures enable reliable, high-throughput analyses.

Objectives and Study Overview

This application note describes the design, packing technology, and performance characteristics of the Agilent ZORBAX Rapid Resolution High Definition (RRHD) SB-C18 threaded column. Key goals include demonstrating high separation efficiency, mechanical stability to 1200 bar, chemical resilience across pH and temperature, and practical operational guidelines.

Methodology and Instrumentation

All evaluations were conducted on an Agilent 1290 Infinity LC system using a ZORBAX RRHD SB-C18 2.1 mm id × 50 mm, 1.8 µm threaded column. Typical conditions:

• Mobile phase: 55 % acetonitrile/45 % water
• Flow rate: 0.55 mL/min
• Temperature: ambient (23 °C); stability tested up to 90 °C at low pH
• Detection: UV at 254 nm
• Sample volume: 2 µL

The column uses a proprietary slurry-loading technique to pack chemically bonded, sterically protected C18 on porous silica microspheres, reducing secondary adsorption of basic analytes.

Main Results and Discussion

Quality-control chromatograms show sharp, symmetrical peaks for uracil, phenol, nitrobenzene, and naphthalene, reflecting high theoretical plate counts. The StableBond C18 phase resists degradation in aggressive mobile phases (pH < 2 or > 6) and supports temperatures up to 90 °C, extending column lifetime. Mechanical tests confirm reliable operation to 1300 bar (20 000 psi) for 2.1 mm and 3.0 mm columns.

Benefits and Practical Applications

The ZORBAX RRHD SB-C18 column offers:

• Exceptional speed and resolution for basic, neutral, and acidic compounds without extensive method modification
• Compatibility with aggressive mobile phases and low carry-over for MS detectors
• Extended lifetime under high pressure, temperature, and variable pH conditions
• Versatility for pharmaceutical assays, environmental screening, and QA/QC workflows

Future Trends and Potential Applications

Advances in UHPLC are driving demand for sub-2 µm stationary phases that combine mechanical robustness with chemical stability. Future developments may include further reduction of particle size, novel surface chemistries for even lower bleed in MS applications, and enhanced column formats for multi-dimensional separations.

Conclusion

The Agilent ZORBAX RRHD SB-C18 threaded column delivers high-definition separations under demanding UHPLC conditions. Its StableBond packing ensures long-term performance across a broad pH and temperature range, making it a versatile choice for high-throughput laboratories.

Reference

• Claessens HA, van Straten MA, Kirkland JJ. J Chromatogr A. 1996;728:259.
• Snyder LR, Kirkland JJ. Introduction to Modern Liquid Chromatography. 2nd ed. John Wiley & Sons; 1979.
• Snyder LR, Kirkland JJ, Glajch JL. Practical HPLC Method Development. 2nd ed. John Wiley & Sons; 1997.