Automated Scouting of Stationary and Mobile Phases Using the Agilent 1290 Infinity II Method Development Solution

Importance of the Topic

Method development in liquid chromatography often requires evaluating multiple stationary phases solvents gradients and temperatures to achieve optimal separation of diverse compounds in pharmaceutical food and environmental analysis laboratories. Automated scouting can accelerate this process reduce manual workload and improve reproducibility.

Objectives and Study Overview

The aim of the study is to demonstrate how the Agilent 1290 Infinity II Method Development Solution together with Method Scouting Wizard software automates the scouting of stationary and mobile phases. The overview covers hardware configuration of multicolumn thermostat and flexible pump as well as software guided generation of comprehensive scouting sequences.

Methodology and Instrumentation

The Method Development Solution integrates the following modules

• Agilent 1290 Infinity II Flexible Pump with two solvent selection valves
• Agilent 1290 Infinity II Multicolumn Thermostat with quick change valve for up to eight columns
• Agilent 1290 Infinity II Multisampler
• Agilent 1290 Infinity II Diode Array Detector
• Agilent OpenLAB CDS ChemStation Edition software with Method Scouting Wizard

Up to eight columns and twenty six solvents can be combined in a single experiment. The Scouting Wizard automates creation of sequences including column selection solvent exchange gradient and temperature variations along with flushing equilibration and storage steps.

Main Results and Discussion

Column scouting of seven stationary phases identified ZORBAX SB C18 Eclipse Plus C8 Eclipse Plus C18 Extend C18 and Bonus RP as top performers under a generic gradient. Solvent scouting on Eclipse Plus C18 at 40 degrees revealed that a ten millimolar ammonium bicarbonate buffer with acetonitrile gave the best resolution. Temperature scouting showed that raising temperature to 60 degrees further improved selectivity. Data analysis using preconfigured bubble plots for peak count and summed resolution enabled rapid identification of optimal conditions. The best separation was achieved on ZORBAX Eclipse Plus C18 at 60 degrees using water and acetonitrile.

Benefits and Practical Applications

• Substantial reduction of method development time through automation
• High reproducibility of retention and selectivity
• Ability to explore over a thousand experimental conditions in a single sequence
• Streamlined data analysis with intelligent reporting templates

Future Trends and Opportunities

Integration of machine learning algorithms could enable adaptive optimization of chromatographic conditions in real time. Expansion of solvent and stationary phase libraries and coupling with mass spectrometry detectors may further enhance method development workflows. Cloud connectivity could support remote monitoring and collaborative method sharing across laboratories.

Conclusion

The combination of Agilent 1290 Infinity II hardware and Method Scouting Wizard software offers a powerful automated platform for rapid comprehensive and reproducible scouting of chromatographic conditions. This solution simplifies method development accelerates time to results and supports high quality analysis in regulated and research environments.

Reference

• Schneider S Performance Characteristics of the Agilent 1290 Infinity II Multicolumn Thermostat Agilent Technologies Technical Overview publication number 5991-5533EN 2015