Agilent 1290 Infinity II LC with ISET

Importance of Topic

Method transfer between ultrahigh performance liquid chromatography and conventional LC platforms is critical for laboratories in pharmaceutical, environmental, and quality control settings. Ensuring consistent retention times and reproducible separations accelerates workflow, reduces revalidation efforts, and supports regulatory compliance.

Objectives and Overview of the Study

This technical overview presents a streamlined workflow for automated method scouting on an Agilent 1290 Infinity II LC with Intelligent System Emulation Technology and subsequent transfer to an Agilent 1260 Infinity LC equipped with an autosampler and integrated column compartment. The study evaluates retention time precision and the accuracy of method transfer with and without ISET.

Methodology and Instrumentation

• UHPLC platform: Agilent 1290 Infinity II with High-Speed Pump, Multisampler with cooler, Multicolumn Thermostat, and Diode Array Detector equipped with a 10 mm Max-Light cell.
• HPLC platform: Agilent 1260 Infinity with Binary Pump, Standard Degasser, Autosampler with integrated column compartment and sample cooler, and Diode Array Detector with 10 mm Max-Light cell.
• Mobile phases: LC-grade water and acetonitrile; gradients ranging from 5 to 95 % B; flow rates 1.0 mL/min for UHPLC and 1.2 mL/min for HPLC; column temperatures at 60 °C; detection at 254 nm with reference at 360 nm.

Main Results and Discussion

Fast UHPLC separations (2.5 min) on the 1290 Infinity II achieved retention time RSDs below 0.1 %. Conventional HPLC runs (30 min) on both platforms delivered RSDs under 0.03 %. Without ISET, retention times on the 1290 II deviated by up to 9 % compared to the 1260. Implementation of ISET reduced deviations to below 1 %, demonstrating seamless transfer and excellent agreement of retention times across systems.

Benefits and Practical Applications

• Rapid exploration of stationary and mobile phase combinations under UHPLC conditions.
• Reliable retention time matching between high-throughput and conventional LC instruments.
• Reduced method revalidation when moving methods between labs or production.
• Enhanced productivity in pharmaceutical development, QC laboratories, and contract testing.

Future Trends and Possibilities

Advances may include integration of machine learning algorithms to predict optimal phase and gradient conditions, expansion of ISET compatibility to additional LC-MS platforms, and cloud-based method sharing to further streamline global collaborations and regulatory submissions.

Conclusion

The combination of Agilent 1290 Infinity II Method Development Solution and ISET enables fast automated method scouting and accurate transfer to an Agilent 1260 Infinity LC with minimal retention time deviation. This approach enhances efficiency and reproducibility in analytical laboratories.

Reference

• Agilent 1290 Infinity with ISET User Manual part number G4220-90314, 2015
• Naegele E and Schneider S Automated Scouting of Stationary and Mobile Phases Using the Agilent 1290 Infinity II Method Development Solution Technical Overview publication number 5991-5934EN, 2015