Effective Analysis Management Achieved by Method Transfer between HPLC and UHPLC

Significance of the topic

The transfer of analytical methods between high performance liquid chromatography and ultra high performance liquid chromatography is essential for modern laboratories seeking both high throughput and consistent data quality. Effective method transfer enables research and development teams to leverage fast UHPLC methods while quality control departments maintain established HPLC protocols. It supports regulatory compliance, cost reduction, and streamlined workflows across different instrument platforms.

Objectives and Study Overview

This report examines bidirectional method transfer between HPLC and UHPLC systems, including transfers among different vendors. It explores the theoretical basis for adjusting parameters, presents case studies on antibiotic analyses, and demonstrates how integrated hardware and software solutions can automate and optimize transfer processes.

Methodology and Instrumentation

Method transfer requires systematic adjustment of key chromatographic parameters to preserve separation performance. Critical factors include:

• Flow rate
• Gradient time program
• System delay volume and sampling rate
• Detector response and injection volume
• Column dimensions and particle size
• Upper pressure limits

Theoretical scaling calculations based on column length, internal diameter, and particle size guide the modification of flow and gradient profiles. The Analytical Conditions Transfer and Optimization function in LabSolutions software automates these calculations, generating compatible methods with minimal manual input.

Instrumentation Used

• Shimadzu Prominence i HPLC system
• Shimadzu Nexera i MT integrated UHPLC/HPLC system
• Shim-pack VP-ODS and Shim-pack XR-ODS II columns
• LabSolutions LC workstation with ACTO (Analytical Conditions Transfer and Optimization)

Key Results and Discussion

Case study 1 improved the analysis speed of a mixture of eleven cephem antibiotics by transferring an HPLC method on Prominence i to UHPLC on Nexera i MT. Flow rate, injection volume, and gradient program were scaled, achieving a sixfold reduction in analysis time while preserving peak resolution and retention order.

Case study 2 generalized a UHPLC method for nine sulfonamide drugs to HPLC conditions. Using ACTO to adjust gradient and system volume, the Nexera i MT system reproduced retention times within 2 percent error when methods were executed on both Prominence i and a third vendor’s system, demonstrating broad compatibility.

Benefits and Practical Applications

• Higher throughput and reduced solvent consumption
• Consistent separation profiles across different departments and instruments
• Simplified regulatory compliance by maintaining relative retention times and resolution
• Automated method generation reduces development time and human error

Future Trends and Potential Applications

Emerging trends include deeper integration of method transfer tools with laboratory information management systems for fully automated workflows. Predictive modeling and machine learning may further refine scaling calculations. Expansion of unified platforms will support a wider range of chromatographic modes and vendors, enabling seamless transfers in complex regulated environments.

Conclusion

Efficient transfer of methods between HPLC and UHPLC enhances laboratory productivity and data consistency. By combining theoretical scaling principles with integrated hardware and automated software tools such as ACTO in LabSolutions, laboratories can achieve rapid, reliable transfer across diverse systems, driving improved analytical performance and operational efficiency.