Seamless method transfer from the Agilent 1200 Series LC System to the Agilent 1260 Infinity LC System

Significance of the Topic

The ability to transfer validated chromatographic methods between instrument platforms is essential in regulated laboratories to maintain compliance with pharmacopoeial standards while avoiding time- and resource-intensive revalidation. Paracetamol and its related impurities serve as a representative model for demonstrating seamless method migration, given the widespread need for reliable quality control in pharmaceutical manufacturing.

Aims and Overview of the Study

This application note describes the transfer of an isocratic liquid chromatography method for paracetamol and its impurities from the Agilent 1200 Series LC System to the Agilent 1260 Infinity LC System. The core objective was to confirm that the new platform meets the European Pharmacopoeia (EP) and United States Pharmacopeia (USP) system suitability criteria without modifying the established method.

Methodology and Used Instrumentation

A consistent batch of mobile phase and the same ZORBAX StableBond-C8 column (4.6 mm×250 mm, 5 µm) was used on both systems. The mobile phase comprised phosphate buffers with tetrabutylammonium hydroxide in methanol. Chromatographic conditions:

• Isocratic flow at 1.5 mL/min
• Column temperature 35 °C
• Injection volume 20 µL
• UV detection at 245 nm with 10 mm pathlength
• Total run time 45 min

Instrumentation details:

• Agilent 1200 Series quaternary pump with external degasser, diode array detector (G1315D), standard autosampler with cooling.
• Agilent 1260 Infinity quaternary pump with integrated vacuum degasser (G1311B), diode array detector (G4212B), advanced autosampler with 10 °C cooling.
• Software: ChemStation B.04.02.

Main Results and Discussion

Retention time shifts between systems were below 1.4% for all peaks. System suitability parameters on the 1260 Infinity matched or exceeded those from the 1200 Series:

• Retention time precision (RSD) < 0.04%
• Area precision (RSD) < 1.2%
• Resolution between paracetamol and Impurity K > 9.6
• Signal-to-noise for late eluting impurity > 1000
• Plate counts > 13 600 for critical peaks
• Peak symmetry and USP tailing within acceptable ranges (symmetry 0.93–1.02; tailing < 1.06)

Overall, the newer system demonstrated improved S/N ratios and enhanced retention time reproducibility by a factor of 1.5–2.

Benefits and Practical Use

Deploying the Agilent 1260 Infinity LC System with the existing method ensures regulatory compliance without full revalidation. Laboratories gain higher confidence in precision and sensitivity, streamline instrument upgrades, and maintain uninterrupted quality control workflows.

Future Trends and Opportunities

Advancements such as higher-efficiency pumps, integrated degassing, faster detectors, and software-driven automated compliance checks will further simplify method transfers. The integration of LC systems with laboratory information management systems (LIMS) and real-time monitoring tools will support continuous validation and predictive maintenance. Miniaturized and micro-flow approaches may expand applicability to high-throughput and resource-limited settings.

Conclusion

The seamless migration of an EP/USP isocratic method for paracetamol from the Agilent 1200 Series to the Agilent 1260 Infinity LC System was successfully demonstrated. All critical system suitability criteria were met, with additional gains in signal-to-noise and retention time precision. This approach minimizes revalidation efforts while upholding strict regulatory standards.

References

1. Wilhelm D. Development, Validation and Comparison of an HPLC Method to Analyze Paracetamol and Related Impurities According to the European Pharmacopoeia (EP) and USP using the Agilent 1120 Compact LC and the Agilent 1200 Series LC System. Agilent Technologies 5990-3739EN, 2009.
2. Rao RN, Narasaraju A. Rapid Separation and Determination of Process-Related Substances of Paracetamol using Reversed-Phase HPLC with Photo Diode Array as Detector. Analytical Sciences. 2006;22:287–292.
3. European Pharmacopoeia 5.0, 2184–2185.
4. European Pharmacopoeia Monograph Paracetamol 01/2005:0049 / 2.2.29.
5. Huber L. Chromatography General Chapter. USP.
6. Agilent Technologies. Understanding Your ChemStation. G2070-91124, 2007.