The Agilent 1260 Infinity Analytical SFC System with Time-of-Flight Mass Spectrometric Detection

Importance of the Topic

Supercritical fluid chromatography (SFC) offers unique selectivity and high-efficiency separations for a wide range of compounds, combining advantages of liquid and gas chromatography. It is particularly valuable in pharmaceutical analysis where fast, high-resolution, and solvent-efficient methods are required.

Objectives and Overview of the Study

This study aimed to automate the development of achiral SFC methods for six pharmaceutical compounds using the Agilent ChemStation Method Scouting Wizard. Four stationary phases and ten mobile phase compositions were screened on an Agilent 1260 Infinity Analytical SFC system coupled to accurate-mass time-of-flight mass spectrometry (TOF MS).

Methodology and Instrumentation

The system comprised an Agilent 1260 Infinity Analytical SFC II Control Module, Binary Pump, II SFC Multisampler, Diode Array Detector with high-pressure cell, and Thermostatted Column Compartment with quick-change valves for up to six columns and twelve solvents.
A 1260 Infinity Isocratic Pump provided make-up flow to the Agilent 6230 TOF LC/MS with Jet Stream ESI. Method Scouting Wizard software generated generic gradients and managed flushing, equilibration, and screening steps.
Key parameters included 2 mL/min flow, 100 bar backpressure, 60 °C column temperature, 1 µL injection, and positive ESI with optimized gas flows and voltages.

Main Results and Discussion

A total of 40 method combinations were evaluated in approximately 11 hours. Three conditions achieved baseline separation and detection of all six target analytes in under three minutes, notably:

• InfinityLab Poroshell 120 HILIC with methanol + 1 % water
• ZORBAX Rx-SIL with pure methanol
• ZORBAX Rx-SIL with methanol + 1 % water

The addition of 1 % water improved peak shapes for certain beta-lactam antibiotics, while retention times varied slightly across solvents and additives, underlining the need for systematic screening.

Benefits and Practical Applications

Automated method scouting significantly accelerates SFC method development by integrating stationary phase and mobile phase optimization. This approach enhances laboratory throughput, reduces time-consuming manual trials, and supports robust quantitation and non-target screening in pharmaceutical R&D and quality control environments.

Future Trends and Opportunities

Emerging trends include AI-driven method optimization, expanded stationary phase chemistries, integration with ion mobility and high-resolution MS, and sustainable solvent strategies. Advancements in automation and data analytics will further streamline SFC workflows.

Conclusion

The application of the Method Scouting Wizard with SFC–TOF MS enabled rapid, comprehensive screening of multiple solvent and stationary phase combinations. The approach identified robust conditions for six pharmaceuticals, demonstrating the efficiency gains and flexibility of automated SFC method development.

References

1. Bieber S.; et al. RRLC-HILIC and SFC with Mass Spectrometry: Polarity Extended Organic Molecule Screening in Environmental (Water) Samples. Anal. Chem. 2017, 89(15), 7907–7914.
2. Agilent Technologies. Strategies for Column and Mobile Phase Selection with the Agilent 1260 Infinity SFC System. Application Note 5990-7147EN.
3. Agilent Technologies. Supercritical Fluid Chromatography with Flexible Injection Volumes at Highest Precision – Performance Evaluation of the Agilent 1260 Infinity II SFC Multisampler in the Agilent Infinity II Analytical SFC Solution. Technical Overview 5991-7623EN.
4. Agilent Technologies. Optimization of Jet Stream ESI Parameters When Coupling Agilent 1260 Infinity Analytical SFC System with Agilent 6230 TOF LC/MS. Technical Overview 5991-4510EN.