Application of an Accurate Mass Database and Library for the Screening and Verification of Extractables and Leachables in Pharmaceutical Drug Products

Importance of the topic

Extractables and leachables analysis is essential to ensure the safety and quality of pharmaceutical products by identifying compounds that may migrate from packaging materials into drug formulations. Robust screening strategies support regulatory compliance and protect patient health.

Study objectives and overview

This work describes the development of an accurate mass personal compound database and library for extractables and leachables (E&L) screening and its application to ophthalmic drug products. The study combines untargeted LC-Q-TOF MS acquisition in positive and negative ion modes with data processing to detect, screen and confirm E&L compounds.

Methodology and data processing workflow

Ophthalmic containers were extracted with a methanol–water mixture at elevated temperature to generate extractables, while drug formulations were thermally stressed to produce leachables. Samples were analyzed on an Agilent 1290 Infinity LC coupled to a 6540 Q-TOF MS using dual-spray Jet Stream electrospray ionization. Full scan MS and data-dependent MS/MS data were acquired in positive and negative modes. Data analysis involved statistical comparison of extract and control scans using Mass Profiler to generate a differential feature list, followed by compound identification via accurate mass and isotopic pattern matching against the E&L library in MassHunter Qualitative Analysis. Unknowns or low-scoring hits were further explored using Molecular Structural Correlator.

Instrumentation used

• Agilent 1290 Infinity UHPLC with binary pump, autosampler, thermostat and column compartment
• Agilent 6540 Q-TOF MS with dual-spray Jet Stream electrospray source
• Agilent 1260 Isocratic Pump for reference mass delivery

Main results and discussion

The constructed library includes accurate mass MS and MS/MS spectra for over 350 compounds and more than 1,000 entries in the final personal compound database. Application to ophthalmic product extracts and stressed formulations yielded confident identification of key extractables such as Erucamide and Irgacure photoinitiators with library match scores above 80. Statistical fold-change analysis highlighted differential features between sample and control sets. Compounds not found in the library were successfully identified via in silico fragmentation matching.

Benefits and practical applications

• Enables rapid screening of a broad range of E&L compounds for regulatory studies
• Provides high confidence in compound confirmation through accurate mass and MS/MS matching
• Supports standardized workflows for container closure evaluation in pharmaceutical development

Future trends and applications

• Expansion of the database to cover additional material types and emerging contaminants
• Integration of targeted acquisition strategies and inclusion lists for enhanced MS/MS coverage
• Adoption of advanced data analytics and machine learning to streamline feature extraction and identification

Conclusion

The accurate mass E&L library combined with a structured data processing workflow demonstrated effective screening and verification of extractables and leachables in ophthalmic drug products. This integrated approach enhances regulatory compliance and supports pharmaceutical quality assurance.

References

• Mire-Sluis A et al. Extractables and Leachables: Challenges and Strategies in Biopharmaceutical Development. BioProcess International. 2011.
• U.S. Department of Health and Human Services FDA. Guidance for Industry: Container Closure Systems for Packaging Human Drugs and Biologics. Rockville MD. 1999.
• Jenke D. Development and Justification of a Risk Evaluation Matrix to Guide Chemical Testing Necessary to Select and Qualify Plastic Components Used in Production Systems for Pharmaceutical Products. PDA Journal of Pharmaceutical Science and Technology. 2015;69:677–712.
• Lateef SS. Extractables and Leachables Detected in Ophthalmic Drug Products. Agilent Technologies Application Note. 2016.