Transfer of an EP method for mebendazole from a Waters Acquity UPLC system to a Vanquish Horizon UHPLC system

Importance of the Topic

Liquid chromatography method transfer is a common requirement in analytical laboratories to ensure consistent, pharmacopeial-compliant results across different instruments and sites.
The ability to accurately replicate a European Pharmacopoeia (EP) monograph method on modern UHPLC platforms is critical for quality control of pharmaceuticals such as mebendazole.

Objectives and Study Overview

This application demonstrates the transfer of an EP monograph HPLC method for mebendazole impurity profiling from a Waters Acquity UPLC system to a Thermo Scientific Vanquish Horizon UHPLC system.
Key aims were to evaluate the impact of instrument dead volume and column thermostatting on retention times, and to scale the method to UHPLC conditions for reduced runtime and solvent consumption.

Methodology and Instrumentation

Sample preparation followed the EP monograph: 5 mg of mebendazole reference standard containing API and impurities A–G dissolved in 5 mL DMF.
Chromatographic conditions were based on a Hypersil GOLD C18 column with a gradient from 20 % to 90 % acetonitrile in ammonium acetate buffer at 40 °C.
Data acquisition and processing used Thermo Scientific Chromeleon CDS 7.2.9.

• Acquity UPLC: Binary Solvent Manager, Sample Manager autosampler with 10 µL loop, high-temperature column heater, tunable UV detector with 10 mm flow cell.
• Vanquish Horizon UHPLC: Binary Pump H, Split Sampler HT (default 25 µL or optional 10 µL loop), Column Compartment H, Variable Wavelength Detector F with 7 mm flow cell.

Main Results and Discussion

Under EP HPLC conditions, both systems yielded equivalent relative retention times and peak areas, with the Vanquish system delivering slightly higher resolution and signal-to-noise despite a shorter optical path.
Absolute retention time shifts of –4 % to –7 % on Vanquish were linked to differences in gradient delay volume and effective column temperature.
Adjusting the Vanquish preheater or oven temperature by 4–7 °C aligned retention times within ±0.5 % of the Acquity system.
Scaling to UHPLC on a 2.1×50 mm, 1.9 µm column reduced run time by 84 % and solvent usage by 90 % while maintaining baseline separation (resolution >2.3) and reproducibility (peak area RSD <0.5 %).

Benefits and Practical Applications

• Seamless transfer of pharmacopeial HPLC methods between different UPLC/UHPLC platforms.
• Flexible thermostatting options to mimic original column temperature conditions.
• Substantial time and solvent savings through UHPLC down-scaling without loss of performance.
• Compliance with EP and USP criteria for system suitability and method robustness.

Future Trends and Opportunities

Advancements in UHPLC system design will further reduce gradient delay volumes and improve temperature uniformity, simplifying cross-platform method transfers.
Integration of automated transfer calculators and CDS tools will streamline revalidation workflows.
The establishment of universal method transfer guidelines may accelerate adoption of standardized assays across laboratories.

Conclusion

This study confirmed that an EP monograph method for mebendazole impurity analysis can be successfully transferred from Acquity UPLC to Vanquish Horizon UHPLC.
Compensation for dead volume and temperature differences enabled equivalent chromatographic profiles, and method scaling achieved dramatic efficiency gains without compromising resolution or reproducibility.

References

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