Enhancing the capabilities of the Agilent 1220 Infinity LC system with the Agilent 1260 Infinity Fluorescence Detector

Importance of the Topic

Polynuclear aromatic hydrocarbons (PAHs) are widespread environmental contaminants known for their mutagenic and toxic properties. Sensitive and selective detection of PAHs is critical for regulatory compliance in environmental and food safety monitoring. Integrating high-pressure liquid chromatography with fluorescence detection enhances analytical performance for trace-level PAH analysis.

Study Objectives and Overview

This Technical Overview demonstrates how an Agilent 1220 Infinity LC system can be enhanced by connecting an external Agilent 1260 Infinity Fluorescence Detector via CAN bus. The focus is on transferring a PAH analysis method between systems and evaluating retention time consistency and detection performance.

Methodology and Instrumentation

• Chromatography system: Agilent 1220 Infinity LC Gradient System (G4290B) with integrated degasser, autosampler, column oven, and variable wavelength detector.
• External detector: Agilent 1260 Infinity Fluorescence Detector (G1321B) connected via CAN bus.
• Control software: Agilent OpenLAB CDS ChemStation Edition Rev. C.01.03.
• Column: ZORBAX Eclipse PAH, 4.6 × 150 mm, 5 µm.
• Mobile phase: A) Water, B) Acetonitrile; gradient from 40 % to 95 % B over 20 min, stop time 30 min, post time 10 min.
• Flow rate: 1.5 mL/min; column temperature: 25 °C; injection volume: 3 µL.
• Detection: variable wavelength detector at 230 nm for acenaphthylene; fluorescence channels with excitation/emission pairs 350/260, 420/260, 440/260, and 500/260 nm.

Main Results and Discussion

Multi-signal fluorescence detection resolved all 16 EPA-610 PAHs, with acenaphthylene monitored by UV. Retention times between the 1220 and 1260 systems differed by no more than 2.2 %, demonstrating robust instrument-to-instrument method transfer. Consistent results were achieved despite differences in mixing principles and column aging over multiple injections.

Benefits and Practical Applications of the Method

• Seamless transfer of legacy HPLC/UHPLC methods across Agilent platforms without revalidation.
• High selectivity and sensitivity for regulatory-level PAH monitoring.
• Modular CAN bus integration enables flexible system upgrades with fluorescence detection.

Future Trends and Potential Applications

Advances in modular connectivity will allow integration of additional detectors and novel column chemistries. Applications may expand to rapid environmental screening, food safety, and high-throughput quality control. Further miniaturization and automation are expected to improve throughput, reduce solvent use, and lower operational costs.

Conclusion

By adding a 1260 Infinity Fluorescence Detector to the 1220 Infinity LC via CAN bus, laboratories can achieve robust, transferable PAH analysis methods with consistent retention times and sensitive detection across different LC platforms.

Reference

• Agilent Application Solution: Analysis of PAHs in soil according to EPA 8310 method with UV and fluorescence detection, publication number 5990-8414EN, 2011.
• Scope of low and high pressure mixing: Comparing the Agilent 1260 Infinity Quaternary and Binary Pumps, publication number 5990-7143EN, 2011.