5.1 The effectiveness of antimicrobial agents incorporated into disinfectants, sanitizers, and antiseptics is measured by their ability to kill microorganisms within a specified contact time. Hence, accurate determination of antimicrobial effectiveness requires complete and immediate inactivation (neutralization) of the antimicrobial agent. Inefficient or incomplete neutralization will permit killing or inactivation of microorganisms to continue beyond the experimental exposure time, resulting in an overestimation of antimicrobial activity.

5.2 The neutralization methods commonly used in antimicrobial effectiveness evaluations are chemical inactivation, dilution, and filtration. All critical parameters of an antimicrobial effectiveness evaluationfor example, media, equipment, microorganism(s), and temperature of solutionsmust be duplicated when evaluating a neutralization procedure to be used.

5.3 The neutralization evaluation must include at least three replications (five replications in Section 9 ) so that a statistical analysis of the recovery data can be performed. The number of replicates used in the evaluation depends on the statistical significance required for the expected results, the variability encountered in the data, and the relative effectiveness of the neutralization procedure.

5.4 A limitation of these evaluation procedures is that they use microorganisms that have not been exposed to an antimicrobial agent. Under experimental conditions, cells exposed to neutralization procedures are likely to be damaged to different degrees by the antimicrobial agent. Sublethal injury may be a factor in recovery, and the role of the neutralization procedure in recovery of injured organisms should be examined. This method is not intended to assess injured organism recovery.

1.1 These test methods are used to determine the effectiveness of procedures and agents for inactivating (neutralizing, quenching) the microbicidal properties of antimicrobial agents, and to ensure that no components of the neutralizing procedures and agents, themselves, exert an inhibitory effect on microorganisms targeted for recovery.

1.2 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.

1.3 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.