Detection and Identification of Extractable Compounds from a Drug Container Closure System Using High Resolution LC/MS and Mass Profiler Software

Význam tématu

Profiling extractable and leachable compounds from pharmaceutical container closure systems is critical for patient safety and regulatory compliance. Impurities migrating from packaging into drug products can pose health risks, and comprehensive analytical coverage is needed to detect substances spanning a wide range of polarities and concentrations. High-resolution liquid chromatography coupled with mass spectrometry (LC/MS) provides the sensitivity and specificity required for untargeted screening of these compounds.

Cíle a přehled studie / článku

The study aimed to develop a generic high-resolution LC/MS method for rapid detection and identification of extractables from an ophthalmic drug container closure system. A standard mixture of nine representative compounds was used to optimize chromatographic separation and mass spectrometric acquisition. Untargeted data processing workflows with MassHunter ProFinder and Mass Profiler Software were implemented to distinguish significant extractables from blank controls and to perform compound identification using a custom extractables database.

Použitá instrumentace

• Agilent 1290 Infinity Binary LC System (Binary Pump G4220A, Autosampler G4226A, Column Compartment G1316A)
• Agilent 6530 Q-TOF Mass Spectrometer with Dual Spray Jet Stream ESI Source (G1959A)
• Agilent 1200 Series ALS Thermostat (G1330B) and DAD Detector (G4212A)
• MassHunter Data Acquisition B.05.01, Qualitative Analysis B.07.00, ProFinder B.06.00, Mass Profiler B.07.00 software

Použitá metodika

Sample preparation involved extracting an empty ophthalmic bottle with 1:1 methanol:water at 55 °C for 72 hours, followed by spiking with a nine-component standard mix. Chromatographic separation used a ZORBAX RRHD Eclipse Plus C8 column (3.0 × 100 mm, 1.8 µm) at 50 °C with a methanol/ammonium acetate gradient (40 % to 100 % B over 8 min) at 0.5 mL/min flow rate. The Q-TOF was operated in All Ions MS/MS mode with three collision energies (0, 15, 40 V) in positive and negative ESI. Data processing consisted of recursive molecular feature extraction and alignment in ProFinder, followed by statistical comparison (fold-change filter ≥ 4, abundance threshold > 3 000) and database searching in Mass Profiler.

Hlavní výsledky a diskuse

The optimized method achieved baseline separation and robust detection of the nine test analytes across both ionization modes, confirming system performance. Untargeted analysis of the bottle extract revealed 66 extractable features significantly above blank levels. Key identifications included plasticizers (dipropyl phthalate), antioxidants (Irganox 1010 degradant), surfactants (sodium ricinoleate), and irritants (diethylene glycol dibenzoate). The combined use of positive and negative mode acquisitions enhanced detection coverage.

Přínosy a praktické využití metody

The described workflow enables rapid, untargeted screening of packaging extractables with high confidence in compound identification. It supports vendor qualification, stability studies, and regulatory submissions by providing comprehensive profiling of potential leachables and automating data analysis for large compound databases.

Budoucí trendy a možnosti využití

Future developments may include expansion of accurate mass databases, incorporation of in silico fragmentation tools, and application of machine learning to improve feature annotation. Integration with automated sample preparation and high-throughput LC/MS platforms can further enhance routine extractables testing in pharmaceutical quality control.

Závěr

A robust LC/Q-TOF method combined with MassHunter ProFinder and Mass Profiler Software successfully identified a diverse set of extractables from an ophthalmic container closure system. The approach provides a generic, high-resolution workflow suitable for pharmaceutical extractables and leachables studies, ensuring patient safety and regulatory compliance.

Reference

• Wong DM, Firor RL. Analysis of Extractable/Leachable Compounds from Transdermal Patches Using GC/MSD Systems. Agilent Technologies Application Note 5991-5605EN, 2015.
• Wong DM, Firor RL. Analysis of Extractable/Leachable Compounds from Plastic Intravenous Bag Sets Using GC/MSD Systems. Agilent Technologies Application Note 5991-5616EN, 2015.
• Wong DM, Firor RL. Analysis of Extractable/Leachable Compounds from Generic Liquid Drug Formulations Using GC/MSD Systems. Agilent Technologies Application Note 5991-5632EN, 2015.
• U.S. Department of Health and Human Services FDA CDER/CBER. Container Closure Systems for Packaging Human Drugs and Biologics: Guidance for Industry. May 1999.
• Norwood DL et al. HPLC and LC/MS Analysis of Pharmaceutical Container Closure System Leachables and Extractables. Journal of Liquid Chromatography & Related Technologies, 2009, 32, pp. 1768-1827.