The Use of ACQUITY UPC2 and the Waters SFC Investigator System for the Analysis, Separation, and Isolation of Canagliflozin and Two Isomeric Impurities

Importance of the Topic

• Analytical separation of isomeric impurities of canagliflozin is crucial for drug quality and safety, as alpha and beta isomers can influence pharmacological activity and regulatory compliance.
• UPC2 and SFC techniques offer faster, highly selective separations compared to conventional reversed-phase chromatography, improving impurity profiling and preparative isolation.

Goals and Study Overview

The study aimed to develop a robust and effective method for separation, isolation, and identification of canagliflozin and its two isomeric impurities using Waters ACQUITY UPC2, the SFC Investigator System, and MS detection. It addresses challenges in resolving alpha and beta sugar moiety isomers in generic API synthesis and supports rapid turnaround for ANDA submissions.

Methodology and Instrumentation

Experiments began with high-throughput screening of various Trefoil chiral column chemistries on the ACQUITY UPC2 system, followed by selection of Trefoil Amylose and Diacel ChiralPak AD-3 columns for optimal resolution. A carbon dioxide–based mobile phase with organic modifiers, Empower 3 software control, and ACQUITY SQD mass spectrometer in positive mode enabled orthogonal detection. Key conditions included a 1.5 mL/min flow rate, 2000 psi ABPR, 45 °C column temperature, and customized gradients for rapid separation.

Main Results and Discussion

The UPC2 screening achieved baseline separation of isomeric peaks in approximately 11 minutes versus 65 minutes by reversed-phase LC. Diacel ChiralPak AD-3 delivered the best resolution of alpha and beta isomers, confirmed by MS signals at m/z 467 [M+Na] and m/z 462 [M+NH4]. Method transfer to the SFC Investigator System reduced runtime to about 15 minutes, optimized at 5 mL/min to support scale-up. Semi-preparative fraction collection yielded recoveries above 90% with high purity.

Benefits and Practical Applications

• Accelerated analysis and reduced runtime compared to traditional LC.
• Lower solvent usage and waste generation.
• Robust QC method with orthogonal MS detection for impurity confirmation.
• Scalability to preparative SFC with high recovery and purity.
• Efficient method development using UPC2 screening to minimize resource consumption.

Future Trends and Potential Applications

Advancements may include broader adoption of UPC2/SFC platforms for diverse chiral separations, integration with automated preparative workflows, expansion to other pharmaceutical compounds, and development of greener, high-throughput impurity isolation processes. Ongoing improvements in column chemistries and detector sensitivity will further enhance performance.

Conclusion

Combining ACQUITY UPC2 screening and SFC Investigator preparative separations provides a rapid, high-resolution workflow for the analysis and isolation of canagliflozin isomers. This integrated approach supports efficient QA/QC operations and scalable purification in pharmaceutical research and manufacturing.

References

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