Robustness of Eclipse PAH Columns for HPLC Analysis of Polycyclic Aromatic Hydrocarbons

Significance of the Topic

Polycyclic aromatic hydrocarbons (PAHs) are environmental pollutants of concern in air, water, and food matrices. Reliable HPLC methods for PAH profiling are essential for regulatory compliance, risk assessment, and quality control. Columns that deliver high resolution, reproducibility, and long lifetime enable consistent results across laboratories and sample types.

Objectives and Study Overview

This application note evaluates the robustness of Agilent’s Eclipse PAH columns for routine HPLC analysis of 16 EPA-listed PAHs. Key aims include demonstrating column longevity, batch-to-batch and particle-size reproducibility, and scalability across a range of column dimensions and particle sizes.

Methodology and Instrumentation

An Agilent 1200 Series RRLC system was used with a binary pump (standard or low delay), HiP-ALS autosampler, thermally controlled column compartment, and diode array detector at 220 nm. A certified PAH standard mixture (Agilent PN 8500-6035) was separated using water/acetonitrile gradients. Eclipse PAH columns in various dimensions (2.1–4.6 mm id; lengths 30–250 mm) and particle sizes (1.8, 3.5, 5 µm) were tested.

Key Results and Discussion

• Resolution: All 16 PAHs were baseline resolved (Rs > 2 for critical pairs) even on 250×4.6 mm, 5 µm columns.
• Throughput: Switching to 1.8 µm, 50×2.1 mm columns reduced analysis time from ~26 to ~6.8 minutes while maintaining resolution.
• Longevity: After 5,000 injections on a 2.1×50 mm, 1.8 µm column, retention, selectivity, and efficiency remained stable, with minimal performance loss.
• Batch Reproducibility: Two independent lots of 3.5 µm material showed identical selectivity and retention.
• Scalability: Overlays of 1.8, 3.5, and 5 µm columns of different lengths and diameters confirmed uniform chemistry and predictable method transfer.

Benefits and Practical Applications

Eclipse PAH columns offer:

• Flexible method development via multiple column dimensions and particle sizes.
• Efficient high-throughput screening with minimized solvent consumption.
• Robust performance for environmental and food safety laboratories requiring consistent PAH analysis.

Future Trends and Potential Applications

Continued miniaturization and faster gradients will further reduce analysis time and solvent use. Integrating these columns with tandem mass spectrometry could enhance selectivity and sensitivity for trace PAH detection. Custom gradients and column architectures will enable automated high-throughput workflows in regulatory and industrial settings.

Conclusion

Agilent Eclipse PAH columns deliver reproducible, scalable, and long-lived performance for diverse PAH analyses. Their uniform stationary phase chemistry across particle sizes simplifies method transfer and supports high-throughput environmental and food testing.

References

1. High-Throughput Gradient Optimization by Easily Minimizing Delay Volume, Agilent Publication 5989-6665EN (2007).
2. Appendix A to Part 136, Methods for Organic Chemical Analysis of Municipal and Industrial Wastewater, Method 610 Polynuclear Aromatic Hydrocarbons, US EPA.