DIS/TSS-2 Jan 25, 1979
EFFICACY DATA REQUIREMENTS
When an antimicrobial Agent is intended for a use pattern that is not reflected by the test conditions specified in the Recommended Methods, one or more test conditions specified in the method must be modified and/or supplementary data developed in order to provide meaningful results relative to the conditions of use. The following basic information is critical to the development and submission of appropriate data.
- EXPOSURE PERIOD
- TYPE OF SURFACE
- HARD WATER
- ORGANIC SOIL
- MICROORGANISM SURVIVAL AFTER DRYING ON A HARD SURFACE
- test microorganisms not specified in the method,
- substitution of a porous surface (e.g., porcelain penicylinder, unglazed ceramic tile) for the specified nonporous surface (stainless steel cylinder, glass slide), and/or
- an organic soil load.
- preparation of inoculum,
- application of inoculum to the carrier,
- the time/temperature and relative humidity conditions for drying the microorganisms on the carrier,
- the technique for removal of the microorganisms from the carrier, and
- the specific assay procedure indicating such details as replication, subculture media/diluents, and the incubation time/temperature conditions for the enumeration procedure employed.
- secondary subcultures must be performed to demonstrate that antimicrobial effects were overcome, or
- at the conclusion of the incubation period specified or employed in the method, the primary culture medium with test carrier must be inoculated with approximately 10 microorganisms/ml of the specific culture under test (documented by actual plate counts) and re-incubated for the specified period to demonstrate that the subculture medium was capable of supporting bacterial growth.
- BATCH REPLICATION FOR MODIFIED TESTS
- for basic efficacy claims (e.g., sterilizers, disinfectants, or sanitizers), 2 samples, representing 2 different batches, instead of 3, and
- for supplemental efficacy claims (e.g., fungicides, virucides, or tuberculocides), one sample instead of 2.
All products tested by the recommended methods may be tested at the exposure periods prescribed in those methods. However, if the product is intended for use at exposure periods shorter or longer than those specified in the method, the method must be modified, in a manner acceptable to the Agency, to reflect the deviation in exposure intended. A modification to provide a shorter exposure period is restricted by the manipulative limitations inherent in the method, while a modification to provide a longer exposure period is restricted by the conditions applicable to the use pattern. If a ten-minute exposure period is necessary for the antimicrobial agent to be effective against the test microorganism the product cannot be represented as an "instantly active" product, or cannot be represented as being "effective in 30 seconds, "one minute," or at any time period shorter than 10 minutes. Also, the product cannot be recommended for use in a manner which is inconsistent with the exposure period necessary for effectiveness (as, for example, "Spray on surface, and immediately wipe with clean cloth") unless the standard method has been modified and reflects efficacy under such conditions of use. In any case, the exposure period or manner of use necessary to provide efficacy must be featured prominently on the product label.
When an antimicrobial agent is intended to be effective in treating a hard porous surface, some of the Recommended Methods may be modified to simulate this more stringent condition by substitution of a porous surface carrier (such as a porcelain penicylinder or unglazed ceramic tile) for the non-porous surface carrier (stainless steel cylinder or glass slide) specified in the method. In addition, control data, described below in Supplemental Recommendation No. 6, must be developed to assure the validity of the test results when this modification of the method is employed. In no case may a surface carrier which represents a less stringent condition be substituted for a surface carrier which is specified in the Recommended Method.
The Recommended Methods may be modified to demonstrate the effectiveness of an antimicrobial agent in hard water. The hard water tolerance level may differ with level of antimicrobial activity claimed. To establish disinfectant efficacy in hard water, all microorganisms (bacteria, fungi, viruses) claimed to be controlled must be tested by the appropriate Recommended Method at the same hard water tolerance level.
An antimicrobial agent identified as a "one-step" cleaner-disinfectant, cleaner-sanitizer, or one intended to be effective in the presence of organic soil must be tested for efficacy by the appropriate method(s) which have been modified to include a representative organic soil such as 5% blood serum. A suggested procedure to simulate in-use conditions where the antimicrobial agent is intended to treat dry inanimate surfaces with an organic soil load involves contamination of the appropriate carrier surface with each test microorganism culture containing 5% v/v blood serum (e.g., 19 ml test microorganism culture $ 1 ml blood serum) prior to the specified carrier-drying step in the method. Control data, described below in Supplemental Recommendation No. 6, must also be developed to assure the validity of the test results when this modification is incorporated into the method. The organic soil level suggested is considered appropriate for simulating lightly or moderately soiled surface conditions. When the surface to be treated has heavy soil deposits, a cleaning step must be recommended prior to application of the antimicrobial agent. The effectiveness of antimicrobial agents must be demonstrated in the presence of a specific organic soil at an appropriate concentration level when specifically claimed and/or indicated by the pattern of use. A suggested procedure for incorporating organic soil load where the antimicrobial agent is not tested against a dry inanimate surface, such as the AOAC Fungicidal Test involves adding 5% v/v blood serum directly to the test solution (e.g., 4.75 ml test solution + 0.25 ml blood serum) before adding 0.5 ml of the required level (5 X 106 /ml) of conidia.
The Recommended Methods are designed to demonstrate efficacy of a freshly prepared antimicrobial solution intended for a single application. When the same use solution is intended for repeated applications, testing must be conducted in accordance with a test protocol specially designed to demonstrate retention of the claimed level(s) of antimicrobial activity in the use solution after repeated microbial and other appropriate challenges (such as supplemental recommendations indicated above) and stress conditions (such as an inadvertent or incidental dilution inherent in the use pattern) over the period of time or number of times specified in the directions for use.
Quantitative determinations of the viable microbial concentration on the untreated control carrier after drying are required in order to determine the validity of the test results obtained with treated carriers when the Recommended Methods are modified to include such elements as:
The detailed protocol for this testing must include:
The test results must include the individual counts obtained by the method.
For each antimicrobial product, procedures must be employed that will preclude residual effects of the active ingredient(s) in the subculture medium. A specific medium capable of neutralizing the antimicrobial effects of a product (whenever one is known) should be employed prior to the microbiological assay. Some of the Recommended Methods rely solely upon the selection of an appropriate subculture medium to neutralize the antimicrobial effects of certain general types of chemical compounds (active ingredients). However, to document absence of residual effects of the active ingredient(s) in the subculture medium, the following testing is necessary:
Where the required batch replication has already been performed and accepted for a product registration with unmodified tests by the Recommended Methods, additional testing at the same use concentration under modified conditions (e.g., different exposure period, presence of organic soil or hard water, porous surface carrier, etc.) may be conducted with reduced batch replication, as follows: