Using a vacuum jacketed column with a multi reflecting time-of-flight MS to increase extractables analysis throughput whilst maintaining identification confidence

Importance of the Topic

Extractables screening by liquid chromatography–mass spectrometry is essential for ensuring the safety and quality of pharmaceutical packaging and other materials in contact with sensitive products. Traditional methods rely on long organic gradients and high solvent use, increasing cost and environmental impact. Accelerating these analyses while preserving peak capacity and identification confidence is critical for high-throughput laboratories and regulatory compliance.

Study Objectives and Overview

This study evaluates the combination of a vacuum jacketed column with a multi-reflecting time-of-flight mass spectrometer (Xevo MRT MS) for extractables analysis. The goals are to shorten chromatographic gradients by over 30 percent, maintain or improve peak capacity, and sustain accurate compound identification through high-resolution, fast-scanning MS.

Used Methodology and Instrumentation

• Sample Preparation: Pharmaceutical packaging extracts in isopropanol spiked with a standard extractables and leachables test mix
• Chromatography: ACQUITY PREMIER system with ACQUITY CORTECS C18 columns (1.6 μm, 2.1 × 50 or 100 mm); vacuum jacketed prototype columns wrapped in an insulated chamber to reduce post-column volume and frictional heating
• LC Conditions: Mobile phase A water plus ammonium acetate and formic acid, mobile phase B methanol; gradients from 15 to as short as 1 minute; flow rates of 0.3–0.5 mL/min; column temperature 50–55 °C
• Mass Spectrometry: Xevo MRT MS operating in positive ESI mode, acquisition range m/z 50–1200, scan speeds up to 50 Hz, low/high collision energies to capture precursor and fragment data, mass error filtering ≤ 1 ppm for confident identifications

Key Results and Discussion

Use of the vacuum jacketed column reduced band broadening by minimizing frictional heating and cut post-column volume through closer emitter placement. Compared to a conventional 10 cm UHPLC column, the 10 cm VJC delivered similar results at standard gradients and superior peak capacity and height when gradients were shortened. At 5 cm length, the VJC provided sharper early-eluting peaks and higher base peak intensity on 4.5- to 7.5-minute gradients. Coupled with 50 Hz MRT MS scanning and mass error filtering, the workflow yielded over 250 confident identifications from more than 10 000 detected features.

Benefits and Practical Applications

• Gradient Reduction: Over 30 percent shorter run times reduce solvent consumption, operating costs and environmental footprint
• Enhanced Peak Capacity: Narrower peaks and higher intensities especially for early-eluting compounds improve detectability
• High Confidence Identification: Fast, high-resolution MRT MS scans with tight mass error filtering minimize false positives
• Increased Throughput: Shorter runs and robust data processing enable larger sample batches per day in extractables screening

Future Trends and Applications

Integration of vacuum jacketed columns with emerging high-resolution MS platforms promises further reductions in analysis time and improved sensitivity. Advances in automated data processing and AI-driven spectral libraries will enhance compound annotation. Broader adoption of adiabatic column designs may extend benefits to lipidomics, proteomics and environmental analyses where high-throughput profiling is required.

Conclusion

This work demonstrates that coupling a vacuum jacketed column to a multi-reflecting TOF mass spectrometer significantly enhances extractables analysis throughput while maintaining identification confidence. Shortened gradients, reduced solvent usage, and improved chromatographic performance meet the demands of modern QA/QC and regulatory environments.

References

• Sanig R and Riba I. Analytical Considerations for Extractables Screening With LC-HRMS. Application Note, January 2022.
• Plumb RS et al. High Throughput UHPLC-MS Lipidomics Using Vacuum Jacketed Columns. Journal of Proteome Research 2022;21(3):691-701.
• Fox J et al. Advanced Oligonucleotide Characterization Using Multi-Reflecting TOF Technology. Application Note, April 2025.
• McCullagh M et al. Extractables, Leachables, and Contact Materials: Benefit of Ion Mobility-Enhanced MS Libraries. Application Note, June 2022.