Container/Closure Integrity Testing and the Identification of a Suitable Vial/Stopper Combination for Low-Temperature Storage at -80 C

Abstract

It was recently found that after storage of a live viral vaccine at -80 °C in glass vials closed with rubber stoppers, a phenomenon was revealed which had not been observed before with other viral products stored at -20 °C: overpressure in the vials. As this phenomenon poses a serious safety problem for medical personnel as well as for the product itself, an investigation was initiated to identify the root cause of the overpressure. After exclusion of possible root causes (differences in air temperature or atmospheric air pressure during filling and quality control testing, outgassing from the formulation buffer) the remaining hypothesis involved a possible container closure integrity issue at low temperature. The glass transition temperatures (T(g)) of many rubber stopper formulations are in the range -55 to -70 °C. At storage temperatures below T(g), the rubber stopper loses its elastic properties and there is a risk that the seal integrity of the vial could be compromised. Loss of seal integrity of the vials near storage temperatures of -80 °C would result in an ingress of cold dense gas into the vial headspace. After removal of the vials from storage at -80 °C, the rubber stoppers could regain their elastic properties and the vials would quickly reseal, thereby trapping the ingressed gas, which leads to overpressure in the vial headspace. Nondestructive laser-based headspace analysis was used to investigate the maintenance of container closure integrity as a function of the filling and capping/crimping process, storage and transport conditions, and vial/stopper designs. This analytical method is based on frequency modulation spectroscopy (FMS) and can be used for noninvasive headspace measurements of headspace pressure and headspace gas composition. Changes in the vial headspace composition and/or pressure are a clear marker for vials that have lost container closure integrity. After storage of a live viral vaccine at -80 °C in glass vials closed with rubber stoppers, overpressure in some of the vials was observed, posing a serious safety problem for medical personnel as well as for the product. A working hypothesis to explain this phenomenon involved a possible container closure integrity issue at these low temperatures. The glass transition temperatures (T(g)) of many rubber stopper formulations are in the range -55 to -70 °C. At storage temperatures below T(g), the rubber stopper loses its elastic properties, resulting in compromised seal integrity of the vial and ingress of cold dense gas into the vial headspace. Upon thawing, the rubber stoppers regain their elastic properties and the vials quickly reseal, thereby trapping the ingressed gas, which leads to overpressure in the vial headspace. Nondestructive, laser-based headspace analysis, which is able to detect changes in headspace pressure and gas composition, was used to investigate the maintenance of container closure integrity. Changes in the vial headspace composition and/or pressure are a clear marker for vials that have lost container closure integrity.