Global Footprint
LIMS system

AOAC 966.04

Determine Sporicidal Activity of Disinfectants

Home >> Antibacterial >> AOAC 966.04

AOAC 966.04 – Evaluation of Sporicidal Activity of Disinfectants

AOAC 966.04 provides a test method to evaluate efficacy of water soluble powders and non-volatile liquid products for their sporicidal action.

AOAC 966.04 serves as the standard testing guideline for registration of disinfectants that claim to have sporicidal efficacy as per regulatory requirements set by US Environmental Protection Agency (EPA) and US Food and Drug Administration (FDA).

Test Process of AOAC 966.04

AOAC 966.04 Test Method

  • Peni cylinders and silk sutures are used as test surfaces (carriers) to perform the test.

  • Each carrier is individually inoculated with spore suspension of Bacillus subtilis and Clostridium sporogenes. For AOAC sporicidal tests, inoculum size should be adjusted between 1.0×105 to 1.0×106. spores/ carrier.

  • In the next step, carriers contaminated with test microorganisms are exposed to test products for specific contact times.

  • At the end of contact time, treated carriers are immediately transferred to a recovery medium enriched with neutralizing compounds.

  • Recovery medium is incubated for a period of 21 days. This incubation period allows for a comprehensive qualitative assessment of any potential growth that may still be present after disinfectant treatment.

Importance of AOAC 966.04 Test

Clostridium difficile and Bacillus subtilis are considered one of major contributors for spreading infections and disease outbreaks within hospital settings. Both bacteria can form endospores, allowing them to endure harsh conditions and persist for extended periods. As a part of robust hygiene practices, disinfectants possessing sporicidal action should be employed to effectively eliminate these endospore forming microbes.

Disinfectants labeled with sporicidal claims are registered and approved by regulatory bodies like the U.S. Environmental Protection Agency (EPA) and the U.S. Food and Drug Administration (FDA). AOAC 966.04 method is designed for compliance reporting of sporicidal disinfectants under EPA and FDA regulatory authorities.

At MIS, we offer an extensive portfolio to test sporicidal efficacy of a diverse range of disinfectant products.

Our team comprises highly skilled microbiologists who possess in-depth knowledge and experience in AOAC testing analysis.
Under our AOAC testing portfolio, we also perform AOAC 965.12 test for tuberculocidal activity of disinfectants.

To get a quote on AOAC 966.04 testing, talk to our experts today.


AOAC 966.04 is a sporicidal activity test designed for evaluating efficacy of water soluble powders and non-volatile liquid products. AOAC 966.04 is recommended by regulatory authorities FDA and EPA as part of the registration process of sporicidal disinfectants products.

AOAC 966.04 test is applicable to water soluble powders and non-volatile liquid products intended for disinfection of hard surfaces.

AOAC 966.04 test takes 4-5 weeks to complete.

We test for AOAC 966.04 using the following bacterial strains: Bacillus subtilis and Clostridium sporogenes.

Contact us for more information

    Explore Other Tests

    Leaving Already?

    Did not find the information you were looking for or need something specific!

      Sign up with Us! Enter your email address for a 10% discount!