Agilent 1290 Infinity Quaternary LC – Support of Columns with 2.1 to 4.6 mm ID to 1200 bar

Importance of the Topic

High-performance liquid chromatography systems must accommodate both conventional HPLC workflows and emerging UHPLC methods. Flexibility in pressure, flow, column dimensions, and temperature is essential to address diverse analytical challenges, from routine quality control to ultrafast separations in high-throughput settings.

Study Objectives and Overview

This technical overview demonstrates how the Agilent 1290 Infinity Quaternary LC System supports analytical columns with 2.1 to 4.6 mm inner diameters and pressures up to 1200 bar, while achieving flow rates up to 5 mL/min. A set of eight antioxidant compounds was used to evaluate performance across a wide range of column formats, flow rates, and backpressures.

Methodology and Instrumentation

A comprehensive instrument configuration was applied:

• Agilent 1290 Infinity Quaternary Pump (G4204A)
• Agilent 1290 Infinity Autosampler (G4226A)
• Agilent 1290 Infinity Thermostat (G1330B)
• Agilent 1290 Infinity Thermostatted Column Compartment (G1316C)
• Agilent 1290 Infinity Diode Array Detector (G4212A)
• OpenLAB CDS ChemStation software C.01.04

Mobile phases comprised water (A), acetonitrile (B), methanol (C), and 1 % TFA in water (D). Column temperatures ranged from 25 °C to 100 °C, injection volumes adjusted to column geometry, and detection at 280 nm with reference at 400 nm.

Key Results and Discussion

Flows between 0.5 and 3.8 mL/min generated backpressures from 118 to 1106 bar while maintaining retention time precision with RSD values typically below 0.35 %. The system covers 100 µL/min to 5 mL/min up to 800 bar, tapering to 2 mL/min at 1200 bar. Conventional HPLC conditions (up to 400 bar), fast UHPLC (400–600 bar), and ultrahigh pressures (600–1200 bar) were demonstrated on columns packed with 5 µm, 3.5 µm, 2.7 µm core–shell, and 1.8 µm sub-2 µm particles. Core–shell columns enabled rapid separations at moderate pressures. Separation of all eight antioxidants was achieved in most experiments; the fastest run completed in under 3 minutes on a 4.6×50 mm core–shell column at 3.8 mL/min.

Benefits and Practical Applications

The 1290 Infinity Quaternary LC System provides:

• Seamless switching between traditional HPLC and UHPLC modes
• Broad flow and pressure range for method flexibility
• High-throughput capabilities for fast analysis
• Compatibility with varying column formats

These features benefit laboratories requiring both routine QC and ultrafast separations in pharmaceutical, biotechnology, and environmental analysis.

Future Trends and Opportunities

Advances may include automated method scouting across diverse column chemistries, integration with mass spectrometry for enhanced sensitivity, and further reductions in run times through higher pressures and novel stationary phases. Green chromatography approaches will drive solvent reduction and energy-efficient operation.

Conclusion

The Agilent 1290 Infinity Quaternary LC System successfully demonstrated its wide dynamic range, robustness, and precision across a variety of column types and operating conditions, meeting the demands of both conventional HPLC and UHPLC applications.

Used Instrumentation

• Agilent 1290 Infinity Quaternary Pump (G4204A)
• Agilent 1290 Infinity Autosampler (G4226A)
• Agilent 1290 Infinity Thermostat (G1330B)
• Agilent 1290 Infinity Thermostatted Column Compartment (G1316C)
• Agilent 1290 Infinity Diode Array Detector (G4212A)
• OpenLAB CDS ChemStation version C.01.04