Method Transfer from an Agilent 1100 Series LC System to an ACQUITY UPLC H-Class System with Gradient SmartStart Technology: Analysis of an Active Pharmaceutical Ingredient and Related Substances

Significance of the Topic

Ensuring consistent chromatographic performance for active pharmaceutical ingredients and impurities across different instrument platforms is essential for regulated laboratories to maintain data integrity, support modernization efforts, and comply with quality requirements.

Goals and Study Overview

This study presents the transfer of a reversed-phase gradient method for abacavir and its related substances from an Agilent 1100 Series HPLC to an ACQUITY UPLC H-Class System. The objective was to replicate separation quality and system suitability without manual gradient modifications by leveraging gradient SmartStart Technology to adjust for differing dwell volumes.

Methodology and Instrumentation

• Column: CORTECS C18, 2.7 µm, 4.6 × 75 mm
• Mobile phase pre-heater: passive (Agilent) vs active (UPLC)
• Dwell volume measurement: 1290 µL (Agilent 1100) and 375 µL (UPLC H-Class)
• Gradient SmartStart Technology: 915 µL after injection delay entered to compensate volume differences
• Data system: Empower 3 FR2

Main Results and Discussion

• Retention times on UPLC were within 3% (≤0.2 min) of the HPLC values
• Relative retention times deviated by ≤0.01%
• USP resolution for the critical API-impurity pair ≥2.5 on both systems
• Peak area repeatability (%RSD) <3% on both instruments; lower on UPLC

Benefits and Practical Applications

Gradient SmartStart Technology eliminated the need for manual gradient table edits and potential method revalidation. The transferred UPLC method maintained comparable separation, delivered improved precision, and enabled faster runtimes, supporting efficient QA/QC workflows in pharmaceutical laboratories.

Future Trends and Potential Applications

Advancements in automated method transfer and dwell volume compensation will accelerate adoption of UHPLC for legacy HPLC assays. Integration with high-throughput data platforms, further miniaturization of systems, and application to diverse compound classes are anticipated to enhance analytical productivity and regulatory compliance.

Conclusion

The successful migration of the abacavir impurity assay demonstrates that gradient SmartStart Technology can preserve chromatographic performance while simplifying method implementation. This approach facilitates seamless modernization of analytical protocols with minimal adjustment.

References

1. United States Pharmacopeia Chapter <621> Chromatography. USP 37–NF 32 S2; 2014.
2. Waters Corporation. CORTECS Columns Applications Notebook; 2013.
3. Waters Corporation. Protocol for Gradient Delay (Dwell Volume) Measurement; 2013.
4. Waters Corporation. Gradient SmartStart Technology Guide. ACQUITY UPLC H-Class System; 2010.
5. Dolan JW. System Suitability. LC·GC Europe. 2004;17(6).