Comprehensive extractables analysis using novel automated parallel extraction and concentration coupled with a multi-detector LC/UV/CAD/HRAM Orbitrap MS system

Significance of the Topic

Extractables and leachables testing plays a crucial role in pharmaceutical development and medical device qualification by assessing potential contaminants that may migrate from packaging materials or single-use systems into the product. Automated and reproducible extraction workflows help to reduce solvent consumption, minimize hands-on time, and improve data quality for regulatory compliance.

Objectives and Study Overview

This study evaluates the performance of the Thermo Scientific EXTREVA ASE Accelerated Solvent Extractor system for automated parallel extraction and on-line preconcentration of extractables from a polypropylene twist-off port. The goal was to develop a fast, reproducible extraction protocol and to demonstrate the utility of a multi-detector LC platform (UV, CAD, HRAM MS) for confident detection, semi-quantitation, and identification of unknown extractables.

Methodology and Instrumentation

Extraction was performed on an EXTREVA ASE system with 50:50 water/isopropanol solvent at 110 °C. Flow rates of 0.2 mL/min (35 min) and 0.35 mL/min (20 min) were compared to optimize analyte recovery. Parallel extraction of up to four samples and automated concentration to 1 mL via vacuum, nitrogen flow, and heating were implemented. A static overnight extraction served as a benchmark. The analysis utilized a Vanquish Duo UHPLC system with inverse gradient, UV diode array detector, Charged Aerosol Detector, and a Thermo Scientific Orbitrap Exploris 120 mass spectrometer with polarity-switching data-dependent MS2. Data processing was conducted in Chromeleon CDS and Thermo Scientific Compound Discoverer with mzVault and mzCloud spectral libraries.

Main Results and Discussion

• The lower flow rate (0.2 mL/min) at 110 °C yielded higher analyte concentrations, establishing optimal conditions.
• Triplicate extractions demonstrated excellent reproducibility, as shown by overlaid UV chromatograms.
• Automated evaporation to 1 mL achieved volume precision within 10 % RSD.
• Over ten extractable components exceeded the analytical evaluation threshold (AET) and were semiquantitated using CAD and UV signals.
• High-resolution MS enabled confident annotation of unknowns, including identification of Irganox 1010 via MS2 spectral matching.

Benefits and Practical Applications

• Significant reduction in manual handling and solvent use through automated parallel extraction and online concentration.
• High reproducibility and throughput support efficient extractables profiling in pharmaceutical and medical device labs.
• Multi-detector approach enhances detection sensitivity, quantitation reliability, and compound identification for regulatory studies.
• Adaptable workflow applicable to diverse materials and extractable testing scenarios.

Future Trends and Opportunities

• Integration of artificial intelligence for accelerated data analysis and compound annotation.
• Expansion of solvent systems and sample types, including biologics and complex formulations.
• Automated solvent exchange capabilities to accommodate non-HPLC-compatible solvents.
• Potential coupling with additional detectors (e.g., NMR) for advanced structural elucidation.

Conclusion

The combination of the EXTREVA ASE system and a multi-detector LC platform provides an efficient, reproducible, and high-throughput workflow for extractables profiling. This integrated approach streamlines sample preparation, enhances data quality, and supports compliance with regulatory requirements in pharmaceutical and medical device analysis.

Instrumentation Used

• Thermo Scientific EXTREVA ASE Accelerated Solvent Extractor
• Thermo Scientific Vanquish Duo UHPLC (UV diode array detector, Charged Aerosol Detector)
• Thermo Scientific Orbitrap Exploris 120 mass spectrometer with OptaMax NG HESI source
• Chromeleon CDS and Compound Discoverer software with mzVault and mzCloud libraries

References

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