Utilisation of Ion Mobility Enabled Collisional Cross Section Measurements for the Comparison of Metabolites across Differing Chromatographic Methods

Significance of Topic

Ion mobility enabled CCS measurement offers a means to align metabolites, including isomeric forms, across UPLC and HPLC platforms without reanalysis, improving throughput in drug metabolism profiling.

Objectives and Study Overview

This study evaluated the feasibility of using ion mobility mass spectrometry to correlate and differentiate Nefazodone metabolites across two complementary chromatographic methods (UPLC and HPLC) and to provide structural insights via drift time and CCS data.

Methodology and Instrumentation

The experimental workflow included:

• Incubation of Nefazodone with rat and human hepatocytes followed by protein precipitation.
• Chromatographic separation using Waters Acquity UPLC and Agilent Eclipse Plus HPLC systems with tailored gradients.
• Analysis on a Waters Vion IMS-QTof mass spectrometer employing non-targeted HDMSE and calibrated ion mobility for CCS determination.

Main Results and Discussion

• Detection of 29 Nefazodone metabolites across both methods; 19 metabolites were successfully correlated by accurate mass and CCS.
• High precision of CCS measurements (RSD 0.05–0.42 %) and strong agreement between methods (Δ CCS < 0.5 %).
• Resolution of isomeric metabolites where mass and retention alone were insufficient.
• Drift time alignment effectively filtered background ions, yielding cleaner spectra for confident metabolite identification.

Benefits and Practical Applications

The integration of IMS and CCS enables laboratories to:

• Compare metabolite profiles without repeated analyses when methods change.
• Select suitable chromatographic approaches for sensitivity and speed.
• Enhance identification confidence through orthogonal structural data.

Future Trends and Potential Applications

Anticipated developments include:

• Automation of CCS databases for broad metabolite libraries.
• Integration with machine learning for rapid isomer annotation.
• Expansion to other drug classes and complex biological matrices.

Conclusion

Ion mobility enabled CCS measurements proved reproducible and method-independent, facilitating metabolite correlation across UPLC and HPLC platforms and supporting more efficient workflows in drug metabolism research.

References

Catherine Holdsworth et al. Utilisation of Ion Mobility Enabled Collisional Cross Section Measurements for the Comparison of Metabolites across Differing Chromatographic Methods. DMDG/GMP Open Meeting, 2016.