Agilent ZORBAX Rapid Resolution High Definition 300SB-C8 Threaded Column

Importance of the Topic

ZORBAX RRHD 300SB-C8 represents an advanced UHPLC stationary phase engineered for high-speed, high-resolution reversed-phase separations, particularly of peptides, proteins, and small organic molecules. Its combination of sub-2 µm particles, wide pore structure, and sterically protected C8 chemistry ensures enhanced reproducibility, mechanical stability, and compatibility with aggressive mobile phases, making it critical for laboratories demanding both sensitivity and robustness.

Objectives and Study Overview

This data sheet aims to describe the design features, performance characteristics, operational guidelines, and applications of the Agilent ZORBAX RRHD 300SB-C8 threaded column. It provides users with key parameters for optimal use, maintenance procedures, and typical chromatographic results to support method development and routine analysis.

Methodology and Stationary Phase Design

The column employs 1.8 µm spherical silica particles with a controlled 300 Å pore size, coated with a diisopropyl-protected C8 bonded phase (StableBond chemistry). A high-pressure slurry-loading technique establishes a uniform bed density. The steric protection layer minimizes unwanted adsorption of basic analytes and enhances chemical stability at low pH.

Instrumentation

• UHPLC System: Agilent 1290 Infinity LC or equivalent capable of up to 1200 bar.
• Column: ZORBAX RRHD 300SB-C8, 2.1 mm × 50 mm, 1.8 µm threaded.
• Detector: UV at 254 nm.
• Typical Mobile Phase: 35% acetonitrile / 65% water at 0.5 mL/min, ambient temperature.

Key Results and Discussion

Quality control chromatograms demonstrate high column efficiency and low system dispersion, with baseline separation of probe compounds (uracil, phenol, 4-chloro-nitrobenzene, naphthalene) within two minutes. The packing tolerates pressures up to 1200 bar and temperatures up to 80 °C, although long-term operation is recommended below 1000 bar and under 80 °C for extended lifetime. Stable performance is maintained under low-pH, high-ionic-strength, and ion-pairing conditions.

Advantages and Practical Applications

• Exceptional speed and resolution for peptide mapping, intact protein analysis, and small molecule separations.
• Low background for mass spectrometric detection.
• Chemical stability in aggressive mobile phases (pH < 2, ion-pair agents).
• Mechanical robustness for high-pressure operation.

Future Trends and Applications

Advances in micro- and nano-flow UHPLC, integration with high-resolution mass spectrometry, and multidimensional chromatography will further leverage RRHD column technology. Customized stationary phases and novel surface chemistries may expand the applicability to polar metabolites and complex biopolymers.

Conclusion

The Agilent ZORBAX RRHD 300SB-C8 threaded column combines high-performance packing technology with robust chemical and mechanical properties, offering reliable, high-throughput separations for research and QA/QC laboratories. Its proven performance under challenging conditions ensures its role as a versatile tool in modern analytical workflows.

References

1. H.A. Claessens, M.A. van Straten, J.J. Kirkland, J. Chromatogr. A, 728 (1996) 259.
2. L.R. Snyder, J.J. Kirkland, J.L. Glajch, Practical HPLC Method Development, 2nd ed., John Wiley & Sons, 1997.