Agilent 1290 Infinity Binary LC System with ISET - Emulation of the Waters Alliance 2695 LC system analyzing b-blockers

Significance of the Topic

In pharmaceutical and industrial laboratories, retaining validated chromatographic methods while upgrading equipment reduces costs and regulatory burden. Emulating legacy LC systems on modern platforms ensures consistent data quality and streamlines method migration to high-performance UHPLC instruments.

Objectives and Study Overview

This application note evaluates the capability of Agilent’s Intelligent System Emulation Technology (ISET) on the 1290 Infinity Binary LC to mimic a Waters Alliance 2695 system. Using a published method for β-blockers, the study compares retention times and resolution on both instruments, with and without ISET, to verify compliance with predefined deviation thresholds.

Methodology and Instrumentation

The original β-blocker separation was developed on a Waters Alliance 2695 using a C18 Atlantis T3 column (4.6×150 mm, 5 µm). Mobile phase A was 10 mM ammonium formate at pH 3; B was acetonitrile. The gradient ramped from 5% to 25% B over 10 min, with a flow of 2 mL/min, injection volume of 10 µL, column temperature of 30 °C, and UV detection at 280 nm.

Instrumentation used:

• Agilent 1290 Infinity Binary LC (G4220A pump, G4226A autosampler, G1330B cooler, G1316C column compartment, G4212A DAD)
• Waters Alliance 2695 with dual absorbance detector
• OpenLAB CDS ChemStation v.C.01.04

Main Findings and Discussion

Without ISET, retention times on the 1290 Infinity shifted up to –15.5%. Activating ISET reduced all retention time deviations to below –2.5%, within the ±5% acceptance criterion. Resolution for each β-blocker pair improved on the 1290 system (deviations better than –5%), attributed to lower post-column dispersion in the modern LC architecture.

Benefits and Practical Applications

By emulating non-Agilent systems, laboratories can:

• Avoid revalidation of existing methods
• Maintain data continuity across instrument upgrades
• Leverage higher sensitivity and resolution of UHPLC platforms

Future Trends and Potential Applications

Extending ISET to additional legacy systems will further simplify method migration. Integration with automated method repositories and cloud-based instrument libraries can accelerate transfers. As UHPLC adoption grows, emulation technologies will play a key role in harmonizing workflows and enabling cross-platform method standardization.

Conclusion

Agilent’s ISET on the 1290 Infinity Binary LC effectively reproduces the chromatographic performance of a Waters Alliance 2695 for β-blocker analysis. Retention time deviations and resolution metrics meet stringent acceptance criteria, demonstrating a reliable route for seamless method transfer.

References

1. Waters Application Note, Analysis of β-Blockers on Atlantis T3, January 2009.