Extractables and Leachables Analysis from Laboratory Tubing

Significance of the Topic

The analysis of extractables and leachables (E&L) is critical for ensuring the safety and integrity of pharmaceutical products that contact polymeric materials, governed by regulatory guidance such as USP chapters 1663 and 1664. It addresses non-volatile organic compounds that may compromise drug quality and patient safety.

Objectives and Study Overview

This study presents a comprehensive LC-MS/MS workflow employing SWATH data-independent acquisition combined with a curated E&L-focused library to improve the detection and identification of non-volatile extractables and leachables from laboratory tubing extracts.

Methodology and Instrumentation

• Sample Preparation: Laboratory tubing soaked in acetone to extract potential leachables.
• Chromatography: ExionLC AD HPLC with PDA detection, Phenomenex Luna Omega C18 column, gradient elution at 0.4 mL/min.
• Mass Spectrometry: SCIEX X500R QTOF operated in SWATH mode with ten m/z windows spanning 100–1300 m/z.
• Data Processing: SCIEX OS Analytics software using sample vs control comparison, targeted XIC list (~300 m/z) and non-targeted peak finding.
• Library Search: Custom NIST MS/MS subset (~3,000 spectra) and ChemSpider integration for unknowns via formula generation and in-silico fragmentation.

Main Results and Discussion

• SWATH acquisition provided MS/MS for coeluting, low-abundance analytes, exemplified by cyclohexylamine detection at m/z 100.076.
• Unmatched peaks, such as a component at RT 5.67 (C17H35N2O2), were characterized using ChemSpider and in-silico MS/MS matching.
• Polymer degradants, including nylon-based fragments, were tentatively identified based on molecular formula assignments.
• Optical detection (PDA) verified key phthalate standards but did not reveal additional non-ionizable leachables.

Benefits and Practical Applications

The proposed workflow offers:

• Comprehensive, unbiased fragmentation reducing missed analytes.
• Regulatory compliance with USP chapters 1663/1664.
• Expandable, user-customizable E&L libraries.
• Integration of MS and optical detection for broader compound coverage.

Future Trends and Potential Uses

• Expansion of specialized E&L spectral libraries through collaborative contributions.
• Machine learning approaches for automated fragment assignment and identification.
• Advanced polymer degradant elucidation using predictive modeling.
• Real-time monitoring and automation of DIA workflows for high-throughput screening.

Conclusion

The SWATH-based high-resolution LC-MS/MS workflow on the SCIEX X500R QTOF platform delivers robust detection and confident identification of extractable and leachable compounds, supported by customizable libraries and complementary optical detection, establishing a reliable method for pharmaceutical material safety assessment.

Used Instrumentation

• SCIEX X500R QTOF mass spectrometer with SWATH acquisition.
• ExionLC AD HPLC system with PDA 200–400 nm detector.
• Phenomenex Luna Omega C18 column (1.6 µm, 150×2.1 mm).
• SCIEX OS Analytics software and custom NIST MS/MS library subset.

References

1. Improved Data Quality Using Variable Q1 Window Widths in SWATH Acquisition - Data Independent Acquisition on TripleTOF and X-Series QTOF Systems. SCIEX Technical note RUO-MKT-02-2879-B.