Analytical Method Transfer to Modern UHPLC Instruments

Importance of the Topic

In pharmaceutical quality control, maintaining robust analytical procedures throughout the product lifecycle is essential for regulatory compliance and consistent data quality. The modernization of legacy HPLC methods to ultra-high performance systems (UHPLC) addresses life-cycle management requirements, enhances sensitivity, and reduces system suitability failures.

Objectives and Study Overview

This study developed and applied a systematic workflow to transfer approximately 200 existing HPLC and UHPLC methods onto the Thermo Scientific Vanquish UHPLC platform. The approach aimed to minimize revalidation efforts while meeting regulatory guidance (ICH Q14 and USP <621>) and improving routine analytical performance in development and QC labs.

Methodology and Instrumentation

A two-step process was used:

1. Determine gradient delay volume (GDV) of the originator and target systems, either by calculation (GDV = flow rate × [T50 – 0.5 × gradient time]) or experimental measurement.
2. Adjust system settings (isocratic hold, mixer volume, sample loop) to match or compensate GDV differences, ensuring system suitability criteria are met.

• Thermo Scientific Vanquish Flex UHPLC with quaternary pump and 100 µL injection loop
• Waters Alliance HPLC system
• ChromSword™ and Chromeleon™ CDS software for robustness studies
• Standard solvents: water, methanol, caffeine or acetophenone solutions for delay volume assessment

Main Results and Discussion

• Over 200 methods for small molecules and biotherapeutics were transferred with minimal requalification.
• System suitability improvements included reduced peak tailing (from 2.4 to ~2.1), increased theoretical plates (>52 000 to >55 000), and consistent %RSD (<0.5%) across injections.
• Signal-to-noise ratio for sensitivity tests improved (from ~12 to >17), reducing rejection rates and downtime.
• Intermediate precision studies comparing Vanquish Flex to legacy HPLC showed equivalent or superior reproducibility (RSDs around 0.1–0.3%) and recovery within 98–102%.

Benefits and Practical Applications

• Significant reduction in system suitability failures and related investigations
• Enhanced sensitivity and signal-to-noise for trace impurity detection
• Streamlined change management under cGMP, aligned with ICH Q14 and USP guidelines
• Flexibility to run both legacy HPLC and modern UHPLC methods on a single platform

Future Trends and Potential Applications

Advances in analytical quality by design (AQbD) and software-driven method optimization will further simplify method transfer. Tunable gradient delay volume solutions and modular hardware designs are likely to become standard, enabling rapid adaptation of methods across diverse UHPLC and HPLC systems. Integration with laboratory automation and data-centric platforms will enhance robustness and compliance.

Conclusion

A mechanistic, data-driven approach to analytical method transfer enables efficient modernization of chromatographic assays with minimal revalidation. Implementing this strategy on modern UHPLC platforms improves analytical performance, operational flexibility, and compliance readiness, ultimately accelerating pharmaceutical development and quality control.

References

1. ICH Q14. Analytical Procedure Development, Draft Version, Endorsed 24 March 2022.
2. USP <621>. Chromatography, General Chapter, Revision November 2021.
3. Thermo Scientific Case Study 000566. Method Transfer onto the Vanquish UHPLC Platform.