Validation of a customized combined chemical and microbiological test for qualification of aseptic preparation processes

Abstract

Introduction

To meet legal requirements, an initial and ongoing training program of production staff must be implemented. As part of their initial and periodic qualification to carry out aseptic preparations, a media fill test (MFT) simulating aseptic processes as closely as possible must be carried out. Moreover, and especially when the handled products are toxic, it is also crucial to carry out an analysis of the chemical contamination generated during the aseptic process. For this reason, the Pharmacy Production Department wanted to implement a method combining both the media fill test and the chemical contamination test (MFT/CCT), by coupling a culture medium with a fluorescent tracer. To gain independence and to reduce costs, the department wished to manufacture its own customized bags and vials of culture medium, subsequently referred to as the “MFT kit”, needed to carry out aseptic filling tests.

Methods

The medium chosen for developing the kit was Tryptone-soy broth (TSB) while two sterile options were available for the fluorescence tracer: quinine hydrochloride and fluorescein. A list of all manipulations handled and to be submitted to the MFT/CCT was created. After aseptic compounding, the kits were quarantined for 14 days in an incubator (22.5 °C ± 2.5 °C) in order to check that the sterility was maintained.

Results

Fluorescence was visually checked for both fluorescent tracers solutions after addition of TSB and no change in fluorescence (λ = 366 nm) could be detected. Fertility tests were also carried out and the results showed that the CCT quinine solution inhibited bacterial growth and therefore gave a non-compliant result. The CCT quinine solution was subsequently withdrawn from further development of the MFT/CCT protocol. The fertility test with CCT fluorescein solution was fully compliant. The MFT/CCT protocol developed was submitted to 20 pharmacy technicians in the production department. No post-incubation turbidity was observed after handling. Post-handling UV revelation revealed an average of 2.2 [1.0–4.0] areas with traces per technician. This result reflects the non-compliance of 30% of the technicians who had to repeat the training. The average number of areas with traces among those who successfully passed the test was 1.6.

Discussion/Conclusion

The development of this combined MFT/CCT protocol is promising, and opens up excellent prospects for the training and (re-)qualification of operators involved in aseptic handling. The cost of producing in-house “MFT kits” is much lower than the cost of commercial kits, and also makes it possible to involve teams in the implementation of MFT/CCT. A stability study on the shelf life of the kits will be conducted, with the aim of manufacturing them on a larger scale and making them available at cost price for other hospitals.