Quality-by-Design-Based Method Development Using an Agilent 1290 Infinity II LC

Importance of the Topic

The development of chromatographic methods based on Quality by Design principles ensures robust, efficient, and consistent performance for pharmaceutical analysis. Establishing a design space through systematic screening and optimization facilitates reliable separation of active pharmaceutical ingredients and impurities, while seamless method transfer to conventional HPLC systems supports routine QA/QC requirements.

Objectives and Study Overview

This study aimed to develop a rapid UHPLC method for Amlodipine and its European Pharmacopeia impurities using QbD software under Waters Empower 3 control and then translate the UHPLC method to conventional HPLC systems. Key steps included:

• Screening of sub-2 µm columns, mobile phases, pH, flow rates, and gradient conditions.
• Optimization through multivariate design to define a proven acceptable region.
• Emulation of HPLC system behavior using Agilent Intelligent System Emulation Technology (ISET).
• Verification of method transfer on Agilent 1260 Infinity LC systems.

Methodology and Instrumentation

Chromatographic method development followed ICH Q8(R2) and Q2(R2) guidance. The workflow comprised:

• UHPLC screening with Fusion QbD software under Empower 3 CDS to select the best column chemistry, solvent type, pH, flow rate, and temperature.
• Design of experiments to optimize gradient time, flow rate, oven temperature, and hold time, yielding a robust method with resolution >2.0 between critical impurities.
• Use of a freeware translation calculator to adjust flow, gradient, and injection volume for HPLC columns.
• ISET emulation of HPLC pump mixing and delay volumes on the 1290 Infinity II LC.

Used Instrumentation

• Agilent 1290 Infinity II LC development system with high-speed pump, multisampler, multicolumn thermostat, diode array detector, valve drive, and solvent selection valve.
• Waters Empower 3 CDS with Agilent Instrument Control Framework and ICS driver.
• Fusion QbD Automated LC Method Development Software (version 9.7.1).
• Agilent Intelligent System Emulation Technology (ISET 4).

Main Results and Discussion

Screening identified an Agilent ZORBAX Eclipse Plus C8 sub-2 µm column with acetonitrile and trifluoroacetic acid buffer at pH 2.0 and 1.2 mL/min as optimal. Design space exploration defined a proven acceptable region where system suitability criteria (resolution >2.0) were met in under 3.7 min. Predicted and experimental critical method attributes agreed within ±2 sigma. The optimized UHPLC method was transferred to HPLC columns of 3.5 µm and 5 µm particles. ISET emulation on the 1290 Infinity II produced chromatograms matching those from a 1260 Infinity system within ±5% retention time and resolution criteria. Both HPLC methods exhibited RSDs below 1.1% for retention time, resolution, and peak area.

Benefits and Practical Applications

This workflow delivered a fast, reproducible UHPLC method that seamlessly adapts to legacy HPLC instruments, reducing method development time and ensuring compliance with QA/QC laboratories. Emulation technology eliminates guesswork in method translation and minimizes revalidation efforts.

Future Trends and Possibilities

Advances may include integration of machine learning for automated design space refinement, cloud-based chromatographic method databases, expanded emulation libraries for more instruments, and real-time adaptive acquisition for continuous method improvement.

Conclusion

The combination of QbD methodology, Fusion QbD software, Agilent 1290 Infinity II LC, and ISET-enabled emulation under Waters Empower 3 control proved effective for rapid UHPLC method development and reliable transfer to conventional HPLC systems. This approach enhances analytical productivity and ensures consistent performance across diverse chromatographic platforms.

References

• Metzlaff M Agilent 1290 Infinity II LC with ISET–Emulation of the Agilent 1100 Series LC Through Waters Empower Software Agilent Technologies Application Note 5991-6541EN 2016
• Vinayak AK QbD Based Method Development on an Agilent 1290 Infinity UHPLC System Combined with a Seamless Method Transfer to HPLC Using Intelligent System Emulation Technology Agilent Technologies Application Note 5991-5701EN 2015
• Huesgen AG Fast screening of mobile and stationary phases with the Agilent 1290 Infinity LC and seamless method transfer to an Agilent 1200 Series LC using ISET Agilent Technologies Application Note 5991-0989EN 2012
• European Pharmacopeia 8.7 04/2016:1491
• HPLC calculator Software for chromatographic performance evaluation and HPLC method transfer University of Geneva website