AOAC Method 966.04 (Modification) Sporicidal Method to Determine Efficacy of Products Used in Aseptic Filling Applications

Purpose of Test

A modification of the AOAC Sporicidal Activity of Disinfectants test method is used to evaluate water products used in aseptic filling applications for registration with the U.S. EPA.

Summary of Test

In this method, a series of stainless steel cylinders (“carriers”) are inoculated with the spores of Bacillus subtilis and Clostridium sporogenes and are desiccated. The carriers containing the dried spore film are then sequentially immersed into 10 mL of disinfectant and are exposed to the disinfectant for a pre-determined exposure time. After exposure, the carriers are sequentially transferred to a liquid subculture medium specifically selected to neutralize the test substance active and to recover any surviving spores. The carriers are incubated and visually examined for the presence or absence of growth. To demonstrate efficacy with the U.S. EPA, no growth must be observed on any treated carrier.