Simplifying Methods Transfer: Novel Tools for Replicating Your Established Methods on an ACQUITY Arc System

Importance of the Topic

In regulated laboratories and quality control environments, transferring validated HPLC methods between different instrument platforms often results in retention time shifts. These shifts can necessitate time-consuming recalculations and manual gradient adjustments, complicating method harmonization and regulatory compliance.

Objectives and Overview

This application note illustrates how the ACQUITY Arc System, equipped with Arc Multi-flow path technology and Gradient SmartStart, replicates an existing Agilent 1260 Quaternary LC method with minimal adjustments. The goal is to achieve retention time alignment within stringent criteria without manual gradient table edits.

Methodology

A standard mixture containing acetaminophen, caffeine, diphenhydramine, reserpine, sulfadimethoxine, flavone, and diclofenac was prepared in 30:70 water:acetonitrile. Chromatographic conditions matched the legacy method: XSelect CSH C18 column (4.6×250 mm, 5 μm) at 30 °C, flow rate 2.0 mL/min, and a gradient from 15 to 35 % B over 3 min, then to 95 % B over 2 min. UV detection was performed at 260 nm.

Instrumentation

• ACQUITY Arc System with Arc Multi-flow path™ (Flow Path 1, dwell volume ~1.1 mL)
• Agilent 1260 Bio Quaternary LC for comparison (dwell volume ~0.76 mL)
• Column heater CH30-A with active preheating
• 2998 PDA detector with analytical flow cell
• Empower 3 FR2 SR2 chromatography software

Main Results and Discussion

Initial transfer using Flow Path 1 yielded retention times within 3 % of the Agilent system for most analytes, except diphenhydramine (peak 3), which showed a 2.4 % shift (0.10 min). Applying the Gradient SmartStart feature, the gradient start was delayed by 0.05 min after injection without altering the gradient table. This adjustment reduced the retention shift for peak 3 to 0.06 min (1.5 %) and aligned all peaks within 1.4 % of the original method.

Benefits and Practical Applications

• Streamlined method transfer with two injections: one for initial alignment, one for dwell volume fine-tuning
• Elimination of manual gradient table edits and calculation errors
• Compliance with USP <621> guidelines for dwell volume adjustments
• Rapid replication of legacy HPLC methods on modern UPLC platforms

Future Trends and Potential Uses

As laboratories seek automated cross-platform harmonization, features like Multi-flow path and Gradient SmartStart will become standard in chromatographic systems. Integration with cloud-based analytical platforms and machine learning–driven optimization may further reduce method development time and improve global consistency.

Conclusion

The ACQUITY Arc System’s combination of Arc Multi-flow path technology and Gradient SmartStart provides an efficient, two-step workflow to transfer established HPLC methods with high retention accuracy. This approach minimizes manual intervention, maintains regulatory compliance, and accelerates method deployment.

References

• United States Pharmacopeia and National Formulary (USP 37-NF 32 S2); United Book Press, Inc.: Baltimore, MD, 2014; p. 6376.