Analysis and Evaluation of Single-Use Bag Extractables for Validation in Biopharmaceutical Applications

Abstract

This paper describes an approach of extractables determination and gives information on extractables profiles for gamma-sterilized single-use bags with polyethylene inner contact surfaces from five different suppliers. Four extraction solvents were chosen to capture a broad spectrum of extractables. An 80% ethanol extraction was used to extract compounds that represent the bag resin and the organic additives used to stabilize or process the polymer films which would not normally be water-soluble. Extractions with1 M HCl extract, 1 M NaOH extract, and 1% polysorbate 80 were used to bracket potential leachables in biopharmaceutical process fluids. The objective of this study was to obtain extractables data from different bags under identical test conditions. All the bags had a nominal capacity of 5 L, were gamma-irradiated prior to testing, and were tested without modification except that connectors, if any, were removed prior to filling. They were extracted at 40 °C for 30 days. Extractables from all bag extracts were identified and the concentration estimated using headspace gas chromatography–mass spectrometry and flame ionization detection for volatile compounds and for semi-volatile compounds, and liquid chromatography–mass spectrometry for targeted compounds. Metals and other elements were detected and quantified by inductively coupled plasma mass spectrometry analysis. The results showed a variety of extractables, some of which are not related to the inner polyethylene contact layer. Detected organic compounds included oligomers from polyolefins, additives and their degradation products, and oligomers from the fill tubing. The concentrations of extractables were in the range of parts-per-billion to parts-per-million per bag under the applied extraction conditions. Toxicological effects of the extractables are not addressed in this paper.

LAY ABSTRACT: Extractables and leachables characterization supports the validation and the use of single-use bags in the biopharmaceutical manufacturing process. This paper describes an approach for the identification and quantification of extractable substances for five commercially available single-use bags from different suppliers under identical analytical conditions. Four test formulations were used for the extraction, and extractables were analyzed with appropriately qualified analytical techniques, allowing for the detection of a broad range of released chemical compounds. Polymer additives such as antioxidants and processing aids and their degradation products were found to be the source of most of the extracted compounds. The concentration of extractables ranged from parts-per-billion to parts-per-million under the applied extraction conditions.