Chiral Screening for SFC and UHPLC with the Agilent 1260 Infinity II SFC/UHPLC Hybrid System

Significance of the Topic

Ensuring chiral purity of pharmaceuticals is critical for efficacy and safety. Traditional method development for enantioseparation often relies on separate SFC and UHPLC instruments, leading to extended timelines and higher costs. The Agilent 1260 Infinity II SFC/UHPLC Hybrid System integrates both modes on a single platform, enabling rapid, resource-efficient screening of chiral stationary phases and mobile phase conditions.

Objectives and Study Overview

This work aims to illustrate automated chiral method scouting with the Agilent 1260 Infinity II Hybrid System and the Method Scouting Wizard software. A series of racemic drugs—including carprofen, ketoprofen, ibuprofen, and indoprofen—were screened across identical Poroshell 120 chiral columns in both SFC and UHPLC modes to compare separation performance and workflow efficiency.

Methodology and Instrumentation

The hybrid system comprises an SFC control module, SFC binary pump, UHPLC binary/quaternary pump, multisampler, diode array detector with high-pressure flow cell, and up to four multicolumn thermostats fitted with Quick Change valve arrays, permitting up to 32 columns. Switching between SFC and UHPLC modes is automated via InfinityLab Quick Change valves. Method Scouting Wizard in OpenLab CDS generated and executed gradient and isocratic screening methods. Samples were prepared at 1 mg/mL in methanol and analyzed using Poroshell 120 Chiral-CF, CD, V, and T (4.6 × 100 mm, 2.7 µm) columns.

Main Results and Discussion

In SFC mode, methanol (0.1 % TEA/TFA) was the optimal modifier among methanol, ethanol, and isopropanol, enabling baseline separation of carprofen and indoprofen on Chiral-T and CD phases. A 2 mL/min gradient (10–50 % modifier) at 30 °C and 140 bar allowed initial scouting within 3.5 hours. Isocratic optimization on Chiral-T (10 % methanol modifier) achieved complete enantiomer resolution. In UHPLC mode, isocratic runs at 40 % methanol (15 mM ammonium formate, pH 4.0) on Chiral-T provided the best balance of retention and resolution for ibuprofen and indoprofen; ketoprofen remained challenging under tested conditions.

Benefits and Practical Applications

The integrated hybrid system streamlines chiral screening by consolidating SFC and UHPLC on one platform, reducing instrument setup time and solvent consumption. Automated method scouting accelerates lead method identification, supports high-throughput environments, and enhances laboratory productivity in pharmaceutical QA/QC and research settings.

Future Trends and Potential Applications

Advances may include coupling hybrid systems with mass spectrometry for enhanced detection, expansion of automated data analytics and AI-driven method optimization, and further scaling of multicolumn clusters for ultra-high-throughput chiral screening and walk-up applications in regulated environments.

Conclusion

The Agilent 1260 Infinity II SFC/UHPLC Hybrid System, paired with Method Scouting Wizard, offers a versatile, efficient approach for rapid chiral method development. By leveraging identical chiral stationary phases across both modes, laboratories can achieve robust enantioseparations with reduced time and resource demands.

References

1. Orthogonal Separations Using the Agilent 1260 Infinity II SFC/UHPLC Hybrid System, Agilent Technologies Technical Overview, publication number 5991-8276EN.
2. Chiral Multicolumn Method Development on the Agilent 1260 Infinity II Analytical SFC System, Agilent Technologies Application Note, publication number 5991-7624EN.
3. Easy Access to Rapid Chiral SFC Method Development for NonChromatographers, Agilent Technologies Application Note, publication number 5994-0011EN.