Method Transfer from an Agilent 1260 Infinity LC to an Agilent 1260 Infinity II LC

Importance of the Topic

Instrument-to-instrument transfer of liquid chromatography methods is essential for regulated pharmaceutical environments and QA/QC laboratories. Ensuring consistent results when migrating from legacy systems to modern LC platforms preserves method validation, boosts laboratory efficiency, and supports regulatory compliance.

Objectives and Study Overview

This application note evaluates the seamless transfer of a validated paracetamol impurity analysis from an Agilent 1260 Infinity LC to an Agilent 1260 Infinity II LC. It compares retention times, resolution, precision, and demonstrates a shortened analysis using the higher pressure range of the newer system.

Instrumentation

• Agilent 1260 Infinity LC: Quaternary Pump (G1311B), High Performance Autosampler with Thermostat (G1367E, G1330B), Thermostatted Column Compartment (G1316A), Diode Array Detector (G4212B)
• Agilent 1260 Infinity II LC: Quaternary Pump (G7111B), Multisampler with Cooler (G7167A), Multicolumn Thermostat (G7116A), Diode Array Detector HS (G7117C)
• Column: Agilent InfinityLab Poroshell 120 EC-C18, 4.6×100 mm, 2.7 µm
• Software: Agilent OpenLAB CDS Version 2.1
• Reagents: LC-grade acetonitrile, ultrapure water, trifluoroacetic acid; paracetamol and impurities A, B, F, H, I, J, K standards

Methodology

• Mobile phases: A = water + 0.1 % TFA, B = acetonitrile + 0.09 % TFA
• Original gradient: 5 % B (0–0.67 min) to 70 % B at 16 min, 1.5 mL/min
• Accelerated gradient: 5 % B (0–0.4 min) to 70 % B at 9.6 min, 2.5 mL/min
• Column temperature 30 °C; sample temperature 6 °C; injection 10 µL
• Detection at 270 nm (4 nm bandwidth) with 395 nm reference (10 nm), data rate 20–40 Hz
• Ten consecutive runs to assess retention time and area precision, and resolution

Results and Discussion

Under standard conditions on the 1260 Infinity LC, retention time RSDs for paracetamol and impurities were <0.05 % and area RSDs <0.7 %; resolution between critical pairs exceeded 3.9. Transferring the method to the 1260 Infinity II LC yielded retention deviations <1 %, improved area precision, and matched resolution. Increasing flow to 2.5 mL/min reduced total run time by ~40 % with only marginal resolution loss, demonstrating robust performance at higher pressures.

Benefits and Practical Applications

• Regulatory compliance via direct method migration to modern hardware
• Consistent performance ensures continuity of QA/QC processes
• Improved precision on advanced detector electronics
• Higher throughput and reduced solvent consumption through accelerated methods

Future Trends and Applications

Continued integration of high-pressure LC platforms with advanced software will drive further adoption of UHPLC for small-molecule analysis. Emerging column technologies, greener mobile phases, automation, and AI-driven method optimization will streamline transfers and harmonize workflows across laboratories.

Conclusion

The Agilent 1260 Infinity to 1260 Infinity II LC transfer for paracetamol impurity analysis is demonstrably equivalent, preserving retention, resolution, and precision. Coupled with faster gradients, laboratories can upgrade instrumentation without revalidating core method parameters, achieving higher throughput and efficiency.

References

1. Agilent Technologies. Agilent 1290 Infinity with ISET, User Manual, Part No. G4220-90314, 2015.
2. ICH Q3A(R2). Impurities in new drug substances, October 25, 2006.
3. Bosch et al. Determination of paracetamol: Historical evolution. Journal of Pharmaceutical and Biomedical Analysis 2006, 42, 291–321.