Straightforward transfer of an EP method for impurity analysis of chlorhexidine from an Agilent 1260 Infinity LC system to a Vanquish Core HPLC system

Importance of the Topic

Reliable transfer of pharmacopeial HPLC methods between different instrument platforms is essential for consistent impurity analysis in pharmaceutical quality control. Efficient method transfer minimizes revalidation effort, ensures compliance with regulatory monographs, and optimizes lab throughput when multiple instruments or sites are involved.

Objectives and Study Overview

This application note demonstrates the transfer of the European Pharmacopoeia monograph method for chlorhexidine impurity analysis from an Agilent 1260 Infinity LC system to a Thermo Fisher Vanquish Core HPLC system. Key goals include verifying equivalent chromatographic performance and showcasing system features that facilitate gradient delay volume (GDV) tuning.

Methodology

The chlorhexidine impurity method employs a C18 column with trifluoroacetic acid–acidified water/acetonitrile mobile phases under a specific gradient program at 1 mL/min, 30 °C column temperature, and UV detection at 254 nm. Standard EP system suitability criteria (resolution ≥3 for certain impurity pairs, peak-valley ratio ≥2) guide performance evaluation.

Used Instrumentation

• Agilent 1260 Infinity Quaternary LC system with standard autosampler, column compartment, and variable wavelength detector.
• Thermo Fisher Vanquish Core Quaternary HPLC system equipped with split sampler CT, column compartment C (passive pre-heater), and VWD C detector.

Key Results and Discussion

Both systems produced highly comparable chromatograms with matching impurity peak profiles, relative retention times, resolution, and peak areas. The Vanquish Core exhibited slightly narrower peaks, yielding marginally improved resolution. Absolute retention times on the Vanquish Core were 0.02–0.22 min earlier due to its lower default GDV. By adjusting the sampler idle volume from 25 µL to 200 µL (or by installing a 200 µL transfer kit loop), retention time deviations were minimized, achieving near-identical alignment with the source system.

Benefits and Practical Applications

• Demonstrates a compliant, straightforward transfer of a compendial HPLC method without parameter revalidation.
• Highlights the Vanquish Core’s GDV-tuning capabilities for rapid retention time matching.
• Ensures consistent impurity profiling across different labs and instrument platforms.

Future Trends and Opportunities

Advancements in HPLC system design—such as enhanced fluidic control, automated volume tuning, and integrated diagnostics—will further streamline method transfer. Incorporation of digital twins or predictive modeling may allow pre-transfer optimization, reducing experimental iterations. Broader adoption of unified data systems will improve cross-platform traceability and compliance.

Conclusion

The EP monograph method for chlorhexidine impurity analysis was successfully transferred from an Agilent 1260 Infinity LC to a Vanquish Core HPLC system, yielding equivalent chromatographic performance. Adjustable gradient delay volume features on the Vanquish Core enabled precise retention time alignment, illustrating a compliant and efficient workflow for pharmacopeial method transfers.

References

• European Pharmacopoeia, 10th Edition (2018), Monograph 0658: Chlorhexidine digluconate solution.
• WHO Model List of Essential Medicines, 21st Edition (2019).
• Chromeleon Chromatography Data System, Version 7.3.