Agilent 1260 Infinity II Prime LC with ISET

Significance of the Topic

The transfer of established chromatographic methods between different liquid chromatography platforms is vital for pharmaceutical quality control. Seamless method migration reduces downtime, cost, and the need for extensive revalidation, ensuring consistent results across laboratories and instruments.

Objectives and Study Overview

This study demonstrates how Agilent’s Intelligent System Emulation Technology (ISET) replicates a Waters Alliance 2695 HPLC method on an Agilent 1260 Infinity II Prime UHPLC system. Using metoclopramide and seven related impurities, the note evaluates retention time and resolution to confirm method fidelity after transfer.

Methodology and Instrumentation

Instrumentation and software employed:

• Waters Alliance 2695 HPLC with Dual Absorbance Detector VWD 2487
• Agilent 1260 Infinity II Prime LC equipped with Flexible Pump (G7104C), Vialsampler (G7129C), Column Compartment, and Diode Array Detector WR (G7115A)
• Control software: Agilent OpenLab CDS 2.2 and Waters DP 2017R2

Chromatographic conditions:

• Column: ZORBAX StableBond C18, 4.6 × 150 mm, 5 µm
• Mobile phase A: 0.1 % formic acid in water; B: 0.1 % formic acid in acetonitrile
• Concave (curve 8) gradient from 8 % to 66 % B over 14 min, then return to initial conditions
• Flow rate: 2 mL/min, column at 25 °C, sample cooler at 8 °C, injection volume 10 µL

Sample preparation:

• Metoclopramide and impurities dissolved to 2 mg/mL in methanol, diluted to 20 ng/µL in water

Key Results and Discussion

Comparative analysis revealed:

• Retention times agreed within 0.9 % between systems, well below the ±5 % acceptance criterion.
• Resolution for most impurity peaks improved on the UHPLC platform, with resolution increases up to 28 %.
• Overlay of 10 consecutive runs on both systems showed consistent peak shape, retention stability (RSD < 0.12 %), and reproducible gradient profiles.

The ability of ISET to emulate concave and linear gradients ensured identical separation performance without method modification.

Benefits and Practical Applications

The adoption of ISET-enabled migration offers:

• Minimized method redevelopment and revalidation efforts
• Preservation of regulatory compliance by replicating legacy system behavior
• Improved throughput and peak resolution on modern UHPLC hardware

Future Trends and Opportunities

Looking ahead, emulation technologies may be extended to:

• Broader instrument families and vendors for universal method transfers
• Automated parameter optimization driven by machine learning
• Real-time method adaptation to accommodate column aging or matrix variability

Conclusion

This application note demonstrates that Agilent ISET seamlessly replicates a Waters Alliance 2695 HPLC method on the 1260 Infinity II Prime LC without altering chromatographic conditions. Excellent agreement in retention and resolution confirms that legacy methods can be migrated efficiently, maintaining data integrity and regulatory compliance.

Reference

1. Gratzfeld-Huesgen A. Agilent 1290 Infinity Binary LC System with ISET – Emulation of the Waters Alliance 2695 LC system analyzing β-blockers. Agilent Technologies Application Note, 5991-1603EN, 2014.
2. Agilent InfinityLab LC with ISET. Agilent Technologies User Manual, Part G7120-90310, 2017.