ASTM E1054-08
(Test Method)Standard Test Methods for Evaluation of Inactivators of Antimicrobial Agents
Standard Test Methods for Evaluation of Inactivators of Antimicrobial Agents
SIGNIFICANCE AND USE
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.
The neutralization methods commonly used in antimicrobial effectiveness evaluations are chemical inactivation, dilution, and filtration. All critical parameters of an antimicrobial effectiveness evaluation—for example, media, equipment, microorganism(s), and temperature of solutions—must be duplicated when evaluating a neutralization procedure to be used.
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.
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.
Note 3—Ideally, all microorganisms used in the antimicrobial effectiveness evaluation should be tested in the neutralization assay. However, representative organisms may be selected for testing, as jud...
SCOPE
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.
Note 1—Knowledge of microbiological and statistical techniques is required for these procedures. These methods are not suitable when testing the virucidal activity of microbicides (see Test Method E 1482).
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.
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Designation: E1054 − 08
StandardTest Methods for
1
Evaluation of Inactivators of Antimicrobial Agents
This standard is issued under the fixed designation E1054; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope 3.1.2 antimicrobial effectiveness evaluation, adj/n—a deter-
mination of microbicidal properties of an antimicrobial agent
1.1 These test methods are used to determine the effective-
by methods, such as Test Methods E645 and E1115.
ness of procedures and agents for inactivating (neutralizing,
quenching)themicrobicidalpropertiesofantimicrobialagents, 3.1.3 CFU/mL (abbrev.)—colony-forming units of a micro-
organism per millilitre of fluid.
and to ensure that no components of the neutralizing proce-
dures and agents, themselves, exert an inhibitory effect on
3.1.4 neutralization, n—a physical or chemical procedure
microorganisms targeted for recovery.
that inactivates or quenches the microbicidal properties of an
NOTE 1—Knowledge of microbiological and statistical techniques is
antimicrobial agent.
requiredfortheseprocedures.Thesemethodsarenotsuitablewhentesting
the virucidal activity of microbicides (see Test Method E1482). 3.1.5 neutralizer effectiveness, adj/n—ability of a neutral-
ization procedure to inactivate or quench the microbicidal
1.2 The values stated in SI units are to be regarded as
properties of an antimicrobial agent.
standard. No other units of measurement are included in this
3.1.6 neutralizer toxicity, adj/n—any inhibitory effects a
standard.
neutralization procedure may have on the survival of a micro-
1.3 This standard does not purport to address all of the
bial population.
safety concerns, if any, associated with its use. It is the
responsibility of the user of this standard to establish appro- 3.1.7 test material control, adj/n—an evaluation of the
priate safety and health practices and determine the applica- activity of a test material in reducing a known population of
bility of regulatory limitations prior to use. microorganisms.
3.1.8 test organism viability, adj/n—the population of a
2. Referenced Documents
challenge microorganism used in a neutralization assay.
2
2.1 ASTM Standards:
3.1.9 viability, n—the ability of a challenge microorganism
E645 Test Method for Efficacy of Microbicides Used in
to form colonies or grow on a nutrient medium.
Cooling Water Systems
3.1.9.1 Discussion—In the context of these test methods,
E1115 Test Method for Evaluation of Surgical Hand Scrub
“viability” is understood to be synonymous with culturability.
Formulations
E1482 TestMethodforNeutralizationofVirucidalAgentsin
4. Summary of Test Methods
Virucidal Efficacy Evaluations
NOTE 2—The neutralization test method selected must be consistent
with the methods of testing used in the antimicrobial effectiveness
evaluation.
3. Terminology
4.1 Neutralization Assay with Recovery on Semi-solid
3.1 Definitions of Terms Specific to This Standard:
Medium—Neutralization assay for antimicrobial effectiveness
3.1.1 antimicrobial, adj—describes an agent that kills or
tests that recover and quantify microbial populations on solid
inactivates microorganisms or suppresses their growth or
(agar) media. This method is appropriate for antimicrobial
reproduction.
agents that are chemically inactivated or diluted to sub-
inhibitorylevelsandperformedentirely in vitroorincludingan
in vivo component to verify neutralization of an antimicrobial
1
These test methods are under the jurisdiction of ASTM Committee E35 on
formulation sampled from the skin of a human volunteer.
Pesticides and Alternative Control Agents and are the direct responsibility of
Subcommittee E35.15 on Antimicrobial Agents.
4.2 NeutralizationAssay with Recovery in Liquid Medium—
Current edition approved April 1, 2008. Published May 2008. Originally
Neutralization assay for antimicrobial effectiveness tests that
approved in 1985. Last previous edition approved in 2002 as E1054 – 02. DOI:
10.1520/E1054-08.
recover surviving microbial populations in liquid media for a
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
growth/no growth determination. This method is appropriate
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
for antimicrobial agents that are chemically inactivated or
Standards volume information, refer to the standard’s Document Summary page on
the ASTM website. diluted to sub-inhibitory levels.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
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E1054 − 08
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This document is not anASTM standard and is intended only to provide the user of anASTM standard an indication of what changes have been made to the previous version. Because
it may not be technically possible to adequately depict all changes accurately, ASTM recommends that users consult prior editions as appropriate. In all cases only the current version
of the standard as published by ASTM is to be considered the official document.
Designation:E1054–02 Designation:E1054–08
Standard Test Methods for
1
Evaluation of Inactivators of Antimicrobial Agents
This standard is issued under the fixed designation E 1054; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (e) indicates an editorial change since the last revision or reapproval.
1. Scope
1.1 These test methods are used to determine the effectiveness of procedures and agents for inactivating (neutralizing,
quenching) the microbiocidal 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.
NOTE 1—Knowledge of microbiological and statistical techniques is required for these procedures. These methods are not applicable tosuitable when
testing with virusesthe virucidal activity of microbicides (see Test Method E 1482).
1.2
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.
2. Referenced Documents
2
2.1 ASTM Standards:
E 645 Test Method for EffectivenessEfficacy of Microbicides Used in Cooling Water Systems
E 1115 Test Method for Evaluation of Surgical Hand Scrub Formulations
E 1482 Test Method for Neutralization of Virucidal Agents in Virucidal EffectivenessEfficacy Evaluations
3. Terminology
3.1 Definitions of Terms Specific to This Standard:
3.1.1 antimicrobial agent—a test formulation, chemical compound, or product designed to prevent the growth of microbes
either by inhibiting growth or destroying the microbe. antimicrobial, adj—describes an agent that kills or inactivates
microorganisms or suppresses their growth or reproduction.
3.1.2 antimicrobial effectiveness evaluation, adj/n—a determination of microbiocidal properties of an antimicrobial agent by
methods, such as Test Methods E 645 and E 1115.
3.1.3 CFU/mLCFU/mL (abbrev.)—colony-forming units of a microorganism per millilitre of fluid.
3.1.4 neutralizer—a procedure or chemical agent used to inactivate, neutralize, or quench the microbiocidal properties of an
antimicrobial agent. neutralization, n—a physical or chemical procedure that inactivates or quenches the microbicidal properties
of an antimicrobial agent.
3.1.5 neutralizer effectiveness—a neutralizer’s ability to inactivate, neutralize, or quench the microbiocidal properties of an
antimicrobial agent. neutralizer effectiveness, adj/n—ability of a neutralization procedure to inactivate or quench the microbicidal
properties of an antimicrobial agent.
3.1.6 neutralizer toxicity, adj/n—any inhibitory effects a neutralization procedure may have on the survival of a microbial
population.
3.1.7 test material control, adj/n—an evaluation of the activity of ana test material in reducing a known population of
microorganisms.
3.1.8 test organism viability—the population or viability of a challenge microorganism used in a neutralization assay. test
organism viability, adj/n—the population of a challenge microorganism used in a neutralization assay.
1
These test methods are under the jurisdiction ofASTM Committee E35 on Pesticides and are the direct responsibility of Subcommittee E35.15 onAntimicrobialAgents
.
Current edition approved May 10, 2002. Published August 2002. Originally published as E1054–85. Last previous edition E1054–91.
1
These test methods are under the jurisdiction ofASTM Committee E35 on Pesticides andAlternative ControlAgents and are the direct responsibility of Subcommittee
E35.15 on Antimicrobial Agents.
Current edition approved April 1, 2008. Published May 2008. Originally approved in 1985. Last previous edition approved in 2002 as E 1054 – 02.
2
For referencedASTM standards, visit theASTM website, www.astm.org, or contactASTM Customer Service at service@astm.org. For Annual Book of ASTM Standards
volume information, refer to the standard’s Document Summary page on the ASTM website.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.
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E1054–08
3.1
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