Agilent 1260 Infinity II Prime LC with ISET - Emulation of the Agilent 1200 Series Quaternary LC for the Analysis of Antihistaminic Drugs

Significance of the topic

The reliable transfer of chromatographic methods between conventional HPLC and modern UHPLC platforms is crucial in regulated laboratories to maintain data consistency and reduce time-consuming revalidation. Differences in system delay volumes and pump mixing behaviors often lead to shifts in retention times and resolution, making seamless method migration a key challenge.

Objectives and study overview

This study evaluates Agilent’s Intelligent System Emulation Technology (ISET) for migrating an isocratic method for five antihistaminic drugs from an Agilent 1200 Series Quaternary LC to an Agilent 1260 Infinity II Prime LC. The goals are to compare retention times, resolution, and precision across systems with and without ISET, and to demonstrate further refinement using ISET fine-tuning.

Methodology and instrumentation

The antihistaminic drug mixture (tripelenamine, chlorpheniramine, tetracaine, promethazine, cetirizine; 150 ng/µL each) was separated on an Agilent ZORBAX SB C18 column (4.6×75 mm, 3.5 µm) using 0.1 % TFA in water (A) and acetonitrile (B). Chromatographic conditions:

• Flow rate: 0.8 mL/min
• Gradient: 10→55 % B in 10 min, stop 12 min, post time 5 min
• Injection: 5 µL, needle wash 3 s (50 % ACN in water)
• Column temperature: 25 °C
• Detection: 232/8 nm, ref off, 20 Hz

Agilent OpenLAB CDS 2.2 controlled both systems.

Used instrumentation

• Agilent 1200 Series Quaternary Pump (G1311A), Autosampler (G1329A), Column Compartment (G1316A), DAD (G1315D)
• Agilent 1260 Infinity II Flexible Pump (G7104C), Multisampler with cooler (G7167A), Multicolumn Thermostat (G7116A), DAD (G7117C)

Main results and discussion

Without ISET, analytes eluted 5–11 % earlier on the 1260 Prime LC. Enabling ISET restored retention times to within 3 % of the original 1200 Series data. Activating fine-tuning (–50 µL delay volume offset) further reduced deviations below 0.5 %.

Precision was assessed over ten consecutive injections. Both systems achieved excellent repeatability: retention time RSDs under 0.06 % and peak area RSDs under 0.14 %, with the 1260 Prime LC showing marginally improved performance. Chromatographic resolution matched or exceeded the original system.

Benefits and practical applications of the method

• Rapid method migration without altering existing chromatographic protocols
• Minimized revalidation time and costs in regulated environments
• Consistent retention times and resolution across HPLC and UHPLC platforms
• Improved precision of retention time and peak area measurements

Future trends and potential applications

Emerging developments may include broader application of ISET to complex gradient and multi-column setups, integration with laboratory information management systems, and AI-driven automated fine-tuning for method optimization. Expansion to other UHPLC and UHPSFC platforms will further streamline cross-system transfers.

Conclusion

ISET enables seamless, reliable migration of chromatographic methods from Agilent 1200 Series Quaternary LC to the 1260 Infinity II Prime LC. With fine-tuning, retention times can be matched to within 0.5 %. The technology preserves resolution and enhances precision, offering substantial operational efficiencies in pharmaceutical and quality-control laboratories.

References

1. Gratzfeld-Huesgen, A. Transferring methods to the Agilent 1290 Infinity LC using Intelligent System Emulation Technology (ISET) – Analysis of metoclopramide hydrochloride and its impurities, Agilent Technologies Application Note, 5990-9692EN, 2014.
2. Agilent InfinityLab LC with ISET, Agilent Technologies User Manual, part number G7120-90310, 2017.