Decontamination and Depyrogenation of an ASEP-TECH® Blow/Fill/Seal SystemChuck Reed, B.Sc/MS, Weiler Engineering; Ryuichi Iwasaki, Noxilizer Japan KK; Tomoyuki Hirose, Noxilizer Japan KK; and David Opie, Ph.D., Noxilizer, Inc.

Results of Decontamination and Depyrogenation in BFS Shroud

Endotoxin Reduction Shows Depyrogenation• Four endotoxin vials were distributed in the shroud in each of 11 cycles• Each endotoxin vial had more than 103 endotoxin units (EU’s)• The observed EU reduction was more than 103 in each cycle, although

cycle parameters were varied during testing (leading to small EU reduction variation)

AbstractTesting has been completed to show the decontamination and depyrogenation of an ASEP-TECH® Blow/Fill/Seal system manufactured by Weiler Engineering, Inc. using nitrogen dioxide (NO2). NO2 has been commercialized for decontamination, sterilization and depyrogenation. The NO2 decontamination and depyrogenationprocess was applied to an ASEP-TECH® Blow/Fill/Seal system. During the testing, cycle parameters were varied to determine optimal process efficacy. Cycle parameters varied included: humidity, NO2 concentration and exposure time. For each exposure cycle, there were nine biological indicators (BI’s) and four endotoxin vials for the demonstration of decontamination and depyrogenation. Each of the BI’s consisted of an inoculated carrier with 106 spores of Geobacillus stearothermophilus. Each endotoxin vial was inoculated with more than 103 endotoxin units (EU). The BI’s and endotoxin vials were widely distributed during testing. The BI results demonstrated complete decontamination, with a consistent 6 log reduction of viable spores. The endotoxin results demonstrated a more than 3 log reduction in EU. These tests utilizing NO2 demonstrate that the ASEP-TECH® Blow/Fill/Seal system can be decontaminated and depyrogenated.

NO2 requires little aeration and results in low (if any) residuals:

• NO2 does not condense on or in product, even in complicated geometries

• Residual NO2 is easily removed via vacuum or air exchanges

• NO2 residuals on a typical syringes and vials yield about 0.01 ppm NO3 in syringes and vials

ConclusionsThe results indicate that this decontamination and depryogenation process offers:• A fast, automated process that can be validated• Increased safety over manual processes• Depyrogenation offers additional safety over

decontamination alone

NO2 as a SterilantNO2 is a rapid and effective sterilant:• Permits rapid sterilization, decontamination or

depyrogenation, depending on cycle parameters• Active at room temperature: 18oC to 30oC• Does not require a lengthy aeration phase• Penetrates packaging and complicated geometries

NO2 exhibits log-linear kinetics of microbial inactivation:

• D-values on the order of 1 minute

• Geobacillus stearothermophilus is the most resistant organism: same as steam and hydrogen peroxide

Endotoxin Log Reduction versus NO2 concentration showing linear dose response

Decontamination Shown with Biological Indicators• Biological indicators (BI’s) had 106 spores of G. stearo.• Nine BI’s used in each cycle• All BI’s were negative (dead); 9 BI’s per cycle x 11 cycles = 99 BI’s

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Based on this, the Blow Fill Seal cycle parameters were:

• NO2 Conc. = 30 mg/L

• Humidity = 55% RH• Decontamination Time = 40 min.

• Aeration Time = 30 min. (During SIP)

Cycle gas concentration versus time using selected parameters

Patent Pending

IMPORTANT WORK FOR THE FUTURE OF BLOW/FILL/SEAL TECHNOLOGY.The critical filling zone of a Blow/Fill/Seal machine is the area comprising the fill system shroud, which typically encompasses the fill needles and electronic modular dosing system. This shroud area has traditionally been steam sterilized or sanitized prior to the start of the production batch. The ability to decontaminate and demonstrate effective endotoxin reduction with a non-aqueous based methodology in an automated process is a new innovation. Particularly advantageous for biologic, protein-based products and heat sensitive products, this new patent-pending application will broaden the use of ASEP-TECH® BFS systems.

• Cycle coincides with the normal CIP/SIP process for cleaning and sterilizing of the product path

• Completely automated process, requiring no human intervention into the filling zone

• Provides a non-aqueous decontamination and depyrogenation of the critical filling zone

• Better than 3-log endotoxin reduction in an easily validated process• Low residuals for high efficacy with sensitive products• Rapid cycle for efficient equipment clearance when running multiple products