Employing UPLC-UV-ToF MSE and the UNIFI Scientific Information System for the Analysis of the Forced Degradation of Glipizide

Importance of topic

The comprehensive analysis of forced degradation products and impurities in pharmaceutical compounds is critical for ensuring drug safety, efficacy, and regulatory compliance. UltraPerformance Liquid Chromatography (UPLC) coupled with high resolution mass spectrometry (HRMS) and integrated data systems enables rapid identification and characterisation of both known and unknown degradation products, supporting robust impurity profiling and quality control in drug development.

Study objectives and overview

This application note describes the forced degradation study of glipizide under acid and thermal stress conditions. The main objectives were:

• To generate degradation products of glipizide by acid catalysis at elevated temperature.
• To perform a comprehensive impurity profiling using UPLC-UV-ToF MSE acquisition.
• To demonstrate data handling, identification, and visualization capabilities of the UNIFI Scientific Information System.

Methodology and instrumentation

Sample preparation involved addition of formic acid to a glipizide solution and incubation at 80 °C, with aliquots taken at 0, 6, 24, 48, 72, and 96 hours. Samples were diluted 1:10 with mobile phase prior to injection.

Chromatographic conditions:

• System: ACQUITY UPLC I-Class with FTN autosampler.
• Column: ACQUITY UPLC BEH C18, 2.1×150 mm, 1.7 µm, at 45 °C.
• Mobile phase A: water + 0.1% formic acid; B: acetonitrile + 0.1% formic acid.
• Gradient: 95% A to 40% A over 7 min, then to 0% A at 8 min, re-equilibration to 95% A by 9 min; flow rate 0.4 mL/min, run time 11 min.
• Injection volume: 0.5 µL; sample temp: 8 °C.

Mass spectrometry conditions:

• Instrument: Xevo G2-XS QTof, ESI+ mode.
• Source temp.: 120 °C; desolvation temp.: 450 °C; desolvation gas: 800 L/hr.
• Acquisition range: m/z 50–1200; scan time: 0.1 s.
• Capillary voltage: 1.0 kV; cone voltage: 25 V.
• MS E functions: low energy 6 eV; high energy ramped 25–45 eV.
• Mass reference: leucine enkephalin at m/z 556.27658.

Data were processed using UNIFI v1.8.2 with pathway profiling workflows, custom calculations, trend plots, and automatic fragment assignment.

Main results and discussion

Two principal degradation products were identified at m/z 379.1074 (C16H18N4O5S) and m/z 321.1020 (C14H16N4O3S), consistent with literature reports. Trend plots showed a time-dependent decrease of parent glipizide and concomitant increase of impurities.

MS E spectra provided simultaneous precursor and fragment ion data. Automatic annotation of fragment ions based on the glipizide structure simplified interpretation of cleavage events. Custom calculations in UNIFI quantified percentage remaining of glipizide and relative response of impurities over the degradation time course.

Benefits and practical applications

• Simultaneous acquisition of full scan and fragment data without precursor selection streamlines method development.
• Integrated PDA and HRMS data enable orthogonal confirmation of impurity identity.
• Custom calculations and trend plotting facilitate rapid visualization of degradation kinetics.
• Automated fragment annotation reduces manual data interpretation effort.
• Robust data management supports storage of spectral libraries and reporting.

Future trends and potential applications

Future developments may include integration of machine learning for automated impurity prediction, expansion to biologics and complex matrices, and high-throughput screening workflows. Enhanced spectral libraries and cloud-based data sharing will further accelerate impurity profiling and regulatory submissions.

Conclusion

The combination of UPLC-UV-ToF MSE acquisition and the UNIFI Scientific Information System provides a powerful platform for forced degradation studies and impurity profiling. This approach enables rapid, comprehensive identification and quantification of degradation products, improving the efficiency and reliability of pharmaceutical analysis.

Reference

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