Universal Analytical Method Development for Various HPLC Systems Using the Agilent 1290 Infinity II Method Development Solution

Significance of the Topic

Modern analytical laboratories must support a wide range of liquid chromatography (LC) platforms from different vendors and generations. Developing robust, transferable methods for diverse instruments is essential to reduce costs, accelerate time-to-result, and ensure consistency across departments or outsourced facilities. A universal workflow that creates one parent method and emulates multiple target systems on-the-fly addresses these challenges by minimizing manual re-optimization and instrument-specific development efforts.

Objectives and Overview of the Study

This study presents a combined approach using the Agilent 1290 Infinity II Method Development Solution with the ChemStation Method Scouting Wizard and Agilent Intelligent System Emulation Technology (ISET) to:

• Develop a single “parent” LC method on the 1290 Infinity II platform.
• Emulate target systems in real time, including the Agilent 1100 Series LC and the Waters Acquity UPLC H-Class.
• Optimize mobile phase, column, gradient, temperature, and flow parameters in a systematic workflow.
• Transfer and confirm method performance directly on each target system without hardware modifications.

Methodology and Used Instrumentation

The workflow comprises three stages: initial screening, refinement, and confirmation. A 15-compound mixture of pesticides and pharmaceuticals (each at 1 mg/mL in acetonitrile) served as the test sample. Key analytical equipment included:

• Agilent 1290 Infinity II Method Development Solution modules (Flexible Pump with ISET, Multisampler, Multicolumn Thermostat, DAD, Single Quad LC/MS).
• Target systems: Agilent 1100 Series LC (quaternary pump, degasser, standard autosampler, column compartment, DAD) and Waters Acquity UPLC H-Class (quaternary solvent manager, sample manager, column manager, TUV detector).
• Software: Agilent OpenLAB CDS ChemStation with Method Scouting Wizard and ISET control; OpenLAB for Waters H-Class control.

Key Results and Discussion

Agilent 1100 Series LC as target:

• Initial screening tested four 4.6×150 mm columns, two solvents (MeOH, ACN), and temperatures of 30–50 °C using a 5–70% organic gradient over 30 min.
• Refinement reduced run time to 17 min, optimized critical peak pair resolution by adjusting gradient slope between 11.5–17 min.
• Transfer to the 1100 system yielded retention time deviations <1% and RSDs <0.03%, based on ten replicates. Total development and evaluation took ≈48 h.

Waters Acquity UPLC H-Class as target:

• Screening of four 2.1×100 mm Poroshell UHPLC columns with 1.9 µm particles revealed pentafluorophenyl (PFP) stationary phase offered distinct selectivity and early elution.
• Method refinement produced a 7.4 min separation from 10–47% ACN at 0.85 mL/min and 40 °C.
• Transfer to the H-Class system delivered retention time deviations <2% and RSDs <0.2% over ten injections. Development and testing required ≈37 h.

Benefits and Practical Applications

The integrated Method Development Solution with ISET enables:

• Single-instrument parent method creation that directly emulates multiple target LCs.
• Automated column, solvent, gradient, temperature, and flow optimization without hardware changes.
• Rapid method transfer and confidence testing on each target system, reducing manual workload and risk of error.

Future Trends and Applications

Expanding this approach could involve:

• Extension to mass spectrometry or two-dimensional LC workflows with real-time emulation.
• Integration of machine-learning algorithms for smarter scouting and parameter prediction.
• Development of shared method libraries across global laboratories to accelerate new product introduction.

Conclusion

The combination of the Agilent 1290 Infinity II Method Development Solution, Method Scouting Wizard, and ISET provides a powerful, generalizable workflow for universal analytical method development. It streamlines screening, refinement, and transfer steps, delivering high reproducibility and minimal retention time deviations across HPLC and UHPLC target systems with significant time savings.

References

1. Huesgen A. G. 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, publication 5991-0989EN, 2012.