Achieving Shortest Run Time and Narrowest Peak Shape Using the Agilent 1290 Infinity II LC with Ultralow Dispersion Kit

Importance of the Topic

High-throughput ultrahigh-performance liquid chromatography (UHPLC) with minimized extracolumn dispersion is critical for modern analytical workflows in pharmaceutical, food, environmental, and clinical laboratories. Achieving sub-30 second separations with narrow, high-efficiency peaks addresses growing demands for rapid sample turnaround, higher sensitivity, and reduced solvent consumption.

Objectives and Overview of the Study

This technical overview describes the development and evaluation of ultrashort isocratic and gradient methods for the separation of nine phenones, employing the Agilent 1290 Infinity II LC system fitted with the Ultralow Dispersion Kit. The study aims to demonstrate minimal peak widths, high plate counts, excellent precision, and enhanced peak capacity at elevated pressures and temperatures.

Methodology and Instrumentation

The following configuration was used:

• Agilent 1290 Infinity II High-Speed Pump (G7120A)
• Agilent 1290 Infinity II Multisampler (G7167B)
• Agilent 1290 Infinity II Multicolumn Thermostat (G7116B)
• Agilent 1290 Infinity II Diode Array Detector (G7117B)
• Agilent Ultralow Dispersion Kit (p/n 5067-5963)
• Ultralow Dispersion Max-Light Cartridge Flow Cell, 10 mm, V(σ)=0.6 µL (G4212-60038)
• Agilent ZORBAX SB-C18, 2.1×50 mm, 1.8 µm column (p/n 857700-902)

Chromatographic conditions:

• Isocratic: water/acetonitrile 45/55, 2.5 mL/min, 1 µL injection, 90 °C, detection 254/4 nm (ref. 360/100 nm)
• Gradient: 55%→95% B in 0.1 min, total run 0.5 min, post time 0.5 min, other settings as above

Main Results and Discussion

Isocratic separation achieved baseline resolution of nine phenones in 27 seconds. Peak widths at half height ranged from 0.002 to 0.010 min, yielding plate counts up to 1,888. Retention time RSDs were below 0.2% and area RSDs below 1.6% at pressures over 1,000 bar and 90 °C.
Gradient separation completed in 16 seconds. Peak widths (W1/2) of 0.002–0.003 min delivered peak capacities up to 100 peaks per 30 s. Precision was outstanding with RT RSDs <0.03% and area RSDs <1%.

Benefits and Practical Applications

• Ultralow dispersion reduces band broadening, maximizing efficiency and sensitivity.
• Sub-30 s run times dramatically increase sample throughput and reduce solvent usage.
• Robust operation at >1,000 bar and 90 °C supports demanding high-pressure applications.

Future Trends and Opportunities

• Extension of ultrafast methods to complex matrices in pharma, food safety, and environmental monitoring.
• Coupling with mass spectrometry for rapid qualitative and quantitative screening.
• Further miniaturization and automation for on-line process control and point-of-care testing.

Conclusion

The Agilent 1290 Infinity II LC with Ultralow Dispersion Kit enables ultrashort isocratic and gradient UHPLC methods with run times under 30 seconds, delivering exceptionally narrow peaks, high plate counts, excellent precision, and peak capacities up to 100. This platform significantly enhances laboratory productivity and analytical performance for high-throughput applications.

Reference

• Schneider, S. Agilent 1290 Infinity LC System – Applications requiring the Agilent Ultralow Dispersion Kit. Agilent Technologies Application Note 5991-0826EN, 2012.
• Schuhn, B.; Schneider, S. Optimizing System Dispersion on the Agilent 1290 Infinity II LC. Agilent Technologies Application Note 5991-5984EN, 2015.
• Agilent A-Line UHPLC Fittings. Agilent Technologies Technical Overview 5991-5525EN, 2015.