Migrating Methods from the ACQUITY™ UPLC™ “Classic” System to the ACQUITY Premier System (FL)

Význam tématu

Lifecycle management of pharmaceutical analytical methods ensures drug safety and expands knowledge in drug product manufacturing. Migrating legacy methods to modern chromatographic systems can improve robustness, sensitivity, and data confidence, particularly in peptide mapping of biotherapeutics where adsorption artifacts compromise impurity detection.

Cíle a přehled studie / článku

The study demonstrates migration of a peptide mapping method from the ACQUITY UPLC™ Classic system to the ACQUITY Premier™ fixed-loop system featuring MaxPeak™ High Performance Surfaces (HPS). Objectives:

• Verify chromatographic equivalence without method adjustments
• Assess the impact of MaxPeak HPS on peak tailing and analyte recovery

Použitá metodika a instrumentace

Samples: mAb tryptic digestion standard at 0.8 mg/mL in 0.1 % formic acid.
Chromatography:

• Systems: ACQUITY UPLC Classic and ACQUITY Premier (fixed-loop)
• Columns: CSH C18 (2.1×100 mm, 1.7 µm) and Premier Peptide CSH C18 (same dimensions)
• Mobile phases: water and acetonitrile with 0.1 % formic acid; gradient 5–95 % B over 25 min
• Flow rate: 0.20 mL/min; column temp: 60 °C; sample temp: 10 °C

Detection:

• UV: ACQUITY TUV at 214 nm
• Mass: ACQUITY QDa (positive, 350–1250 m/z)
• Data software: Empower 3

Hlavní výsledky a diskuse

• Dwell volume: Classic 149.4 µL vs Premier 158.8 µL, enabling direct method transfer without gradient changes
• Retention times and peak areas of 22 peptides matched across systems (slope ~1.00; area differences <0.5 %)
• MaxPeak HPS reduced peak tailing for acidic peptides, improving recovery and sensitivity
• Up to 70 % reduction in tailing enabled detection of deamidated critical quality attribute impurities

Přínosy a praktické využití metody

Migration to ACQUITY Premier fixed-loop provides:

• Seamless transfer of legacy UPLC methods
• Enhanced peak shape and recovery for metal-sensitive analytes
• Improved impurity profiling and lifecycle management of biotherapeutics

Budoucí trendy a možnosti využití

Emerging directions:

• Broader application of high-performance surface technologies in biomolecule separations
• Automated method scaling and development tools
• Integration of fixed-loop designs for reproducible analytical pipelines

Závěr

The ACQUITY Premier System with fixed-loop and MaxPeak HPS achieves direct, robust migration from legacy UPLC methods while enhancing chromatographic performance. It minimizes adsorption artifacts, improves peptide mapping reliability, and supports advanced quality control of biotherapeutics.

Reference

1. Guidance for industry Q10 pharmaceutical quality system. ICH, 2009.
2. P. Bigos, R. Birdsall, Y. Yu. Obtaining Equivalent IEX Chromatographic Performance Through Automated Method Scaling Using Waters Column Calculator. Waters Application Note 720007807, 2022.
3. Chelius D, Rehder DS, Bondarenko PV. Identification and Characterization of Deamidation Sites in the Conserved Regions of Human Immunoglobulin Gamma Antibodies. Anal. Chem. 77: 6004, 2005.