New studies on leachables in commercial scale protein drug filling lines using stir bar sorptive extraction coupled with TD-GC-MS and UPLC/QTOF-MS/MS analytics.

The increasing application of Single-Use Systems (SUSs) in pharmaceutical manufacturing lines poses a potential risk of polymer-related impurities leaching into the process stream and persisting through the manufacturing process. To minimize any potential toxicity and impairment to the product's quality, safety thresholds are strictly regulated and enforced in particular for parenteral solutions. At present, impurities are estimated from extractable profiles, which are generated for each SUS with thermal or static extraction. In this study we employed target leachable-testing by taking samples directly from an industrial filling line probed during real-life processing of three parenteral drugs (n = 2) under actual process-conditions, to estimate the concentration of leachables throughout drug-manufacturing. At five different points, samples were drawn to study the individual impact of SUSs on the leachable accumulation within the drug-filling process. The drug products were examined for leachables using stir-bar-sorptive-extraction (SBSE) with polydimethylsiloxane (PDMS) and ethylene glycol (EG)-PDMS coated stir-bars. Subsequent extraction from the stir-bars and analysis of the substances was performed with TD-GC-MS and solvent-back-extraction (SBE)-UPLC/QTOF-MS/MS analytics. Our study revealed the following main results: 1) Leachables were found in extremely low concentrations, all below toxicological thresholds (highest leachable concentration in the final drug product 1 (DP1): 0.274 ppm < drug specific threshold: 6.0 ppm | DP2: 0.010 ppm < 0.2 ppm | DP3: 0.011 ppm < 0.5 ppm). All compounds identified in the leachables study were found to be non-genotoxic. 2) Most of the leachables (68%) that were found were already observed at the beginning of the filling process, delivered by the API Neither a common source of leaching could be identified within the filling-line nor a specific product influence on quality or quantity of leachables. 3) No leachable increase could be observed over the filling process. On the contrary leachable concentrations declined with 83%, which was partly due to dilution by buffer-feed and to a proven absorption of leachables by filters and silicon tubing.