ASTM E1054-21e1

Standard Practices for Evaluation of Inactivators of Antimicrobial Agents

SIGNIFICANCE AND USE
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 evaluation—for example, media, equipment, microorganism(s), and temperature of solutions—must be duplicated in the performance of selected neutralization procedure.  
5.3 The neutralization evaluation must include at least three replications (five replications in Section 9) so that a statistical analysis of the microbial 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 effect of the neutralization procedure on recovery of injured organisms should be examined. This method is not intended to assess recovery of injured organisms.
Note 3: Ideally, all microorganisms used in the antimicrobial effectiveness evaluation should be tested in the neutralization assay. However, representative organism...
SCOPE
1.1 These test procedures are used to determine the effectiveness of methodologies procedures and materials intended for inactivating (neutralizing, quenching) the microbicidal properties of antimicrobial agents; to ensure that no components of the neutralizing procedures and materials, themselves, exert an inhibitory effect on microorganisms targeted for recovery; and to demonstrate that the antimicrobial chemistry tested is microbicidal.  
1.2 Knowledge of microbiological and statistical techniques is required for these procedures.
Note 1: These methods are not suitable when testing the virucidal activity of microbicides (see Test Method E1482).  
1.3 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.4 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, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.5 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

General Information

• Status: Historical
• Publication Date: 31-Mar-2021
• ICS: 11.080.20 - Disinfectants and antiseptics
• ICS: 71.100.35 - Chemicals for industrial and domestic disinfection purposes
• Technical Committee: E35 - Pesticides, Antimicrobials, and Alternative Control Agents
• Drafting Committee: E35.15 - Antimicrobial Agents

Standards Content (Sample)

NOTICE: This standard has either been superseded and replaced by a new version or withdrawn.
Contact ASTM International (www.astm.org) for the latest information
´1
Designation: E1054 − 21
Standard Practices 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.Anumber in parentheses indicates the year of last reapproval.A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1
ε NOTE—Editorial changes were made throughout in May 2021.
1. Scope E1115Test Method for Evaluation of Surgical Hand Scrub
Formulations
1.1 These test procedures are used to determine the effec-
E1482Practice for Use of Gel Filtration Columns for Cyto-
tiveness of methodologies procedures and materials intended
toxicity Reduction and Neutralization
for inactivating (neutralizing, quenching) the microbicidal
E2756Terminology Relating toAntimicrobial andAntiviral
properties of antimicrobial agents; to ensure that no compo-
Agents
nentsoftheneutralizingproceduresandmaterials,themselves,
exert an inhibitory effect on microorganisms targeted for
3. Terminology
recovery; and to demonstrate that the antimicrobial chemistry
tested is microbicidal.
3.1 Definition of Terms: For definition of terms not listed
below, refer to E2756, Standard Terminology Relating to
1.2 Knowledgeofmicrobiologicalandstatisticaltechniques
Antimicrobial and Antiviral Agents.
is required for these procedures.
NOTE 1—These methods are not suitable when testing the virucidal
3.2 Definitions of Terms Specific to This Standard:
activity of microbicides (see Test Method E1482).
3.2.1 antimicrobial effectiveness evaluation, adj/n—a deter-
1.3 The values stated in SI units are to be regarded as
mination of microbicidal properties of an antimicrobial agent
standard. No other units of measurement are included in this
by methods, such as Practice E645 and Test Method E1115.
standard.
3.2.2 CFU/mL (abbrev.)—colony-forming units of a micro-
1.4 This standard does not purport to address all of the
organism per millilitre of fluid.
safety concerns, if any, associated with its use. It is the
3.2.3 neutralizer effectiveness, adj/n—ability of a neutral-
responsibility of the user of this standard to establish appro-
ization procedure to inactivate or quench the microbicidal
priate safety, health, and environmental practices and deter-
properties of an antimicrobial agent.
mine the applicability of regulatory limitations prior to use.
3.2.4 neutralizer toxicity, adj/n—any inhibitory effects a
1.5 This international standard was developed in accor-
neutralization procedure may have on the survival of a micro-
dance with internationally recognized principles on standard-
bial population.
ization established in the Decision on Principles for the
Development of International Standards, Guides and Recom-
3.2.5 test material control, adj/n—an evaluation of the
mendations issued by the World Trade Organization Technical activity of a test material in reducing a known population of
Barriers to Trade (TBT) Committee.
microorganisms.
3.2.6 test organism viability, adj/n—the population of a
2. Referenced Documents
challenge microorganism used in a neutralization assay.
2
2.1 ASTM Standards:
3.2.7 viability, n—the ability of a challenge microorganism
E645Practice for Evaluation of Microbicides Used in Cool-
to form colonies or grow on a nutrient medium.
ing Water Systems
3.2.7.1 Discussion—In the context of these test methods,
“viability” is understood to be synonymous with cultivability.
1
These practices are under the jurisdiction of ASTM Committee E35 on
Pesticides, Antimicrobials, and Alternative Control Agents and are the direct
4. Summary of Practices
responsibility of Subcommittee E35.15 on Antimicrobial Agents.
NOTE 2—The neutralization test procedure selected must be consistent
Current edition approved April 1, 2021. Published May 2021. Originally
with the methods of testing used in the antimicrobial effectiveness
approved in 1985. Last previous edition approved in 2013 as E1054–13. DOI:
evaluation.
10.1520/E1054-21E01.
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
4.1 Neutralization Assay with Recovery on Semi-solid
contact ASTM Customer Service at [e-mail]. For Annual Book of ASTM
Medium—Neutralization assay for antimicrobial effectiveness
Standards volume information, refer to the standard’s Document Summary page on
the ASTM website. tests that recover and quantify microbial populations on
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1

---------------------- Page: 1 ----------------------
´1
E1054 − 21
samples will contain multiple species of microorganisms (for example,
semi-solid (agar) media. This method is appropriate for anti-
skin flor
...

This document is not an ASTM standard and is intended only to provide the user of an ASTM 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.
´1
Designation: E1054 − 21 E1054 − 21
Standard Test Practices 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
ε NOTE—Editorial changes were made throughout in May 2021.
1. Scope
1.1 These test procedures are used to determine the effectiveness of methodologies procedures and materials intended for
inactivating (neutralizing, quenching) the microbicidal properties of antimicrobial agents; to ensure that no components of the
neutralizing procedures and materials, themselves, exert an inhibitory effect on microorganisms targeted for recovery; and to
demonstrate that the antimicrobial chemistry tested is microbicidal.
1.2 Knowledge of microbiological and statistical techniques is required for these procedures.
NOTE 1—These methods are not suitable when testing the virucidal activity of microbicides (see Test Method E1482).
1.3 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.
1.4 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, health, and environmental practices and determine the applicability of
regulatory limitations prior to use.
1.5 This international standard was developed in accordance with internationally recognized principles on standardization
established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued
by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
2. Referenced Documents
2
2.1 ASTM Standards:
E1115 Test Method for Evaluation of Surgical Hand Scrub Formulations
E1482 Practice for Use of Gel Filtration Columns for Cytotoxicity Reduction and Neutralization
E2756 Terminology Relating to Antimicrobial and Antiviral Agents
3. Terminology
3.1 Definition of Terms: For definition of terms not listed below, refer to E2756, Standard Terminology Relating to Antimicrobial
and Antiviral Agents.
3.2 Definitions of Terms Specific to This Standard:
1
These test practices are under the jurisdiction of ASTM Committee E35 on Pesticides, Antimicrobials, and Alternative Control Agents and are the direct responsibility
of Subcommittee E35.15 on Antimicrobial Agents.
Current edition approved April 1, 2021. Published May 2021. Originally approved in 1985. Last previous edition approved in 2013 as E1054 – 13. DOI:
10.1520/E1054-21.10.1520/E1054-21E01.
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or contact ASTM Customer Service at [e-mail]. 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
1

---------------------- Page: 1 ----------------------
´1
E1054 − 21
3.2.1 antimicrobial effectiveness evaluation, adj/n—a determination of microbicidal properties of an antimicrobial agent by
methods, such as Test Methods E645 and E1115.
3.2.2 CFU/mL (abbrev.)—colony-forming units of a microorganism per millilitre of fluid.
3.2.3 neutralizer effectiveness, adj/n—ability of a neutralization procedure to inactivate or quench the microbicidal properties of
an antimicrobial agent.
3.2.4 neutralizer toxicity, adj/n—any inhibitory effects a neutralization procedure may have on the survival of a microbial
population.
3.2.5 test material control, adj/n—an evaluation of the activity of a test material in reducing a known population of
microorganisms.
3.2.6 test organism viability, adj/n—the population of a challenge microorganism used in a neutralization assay.
3.2.7 viability, n—the ability of a challenge microorganism to form colonies or grow on a nutrient medium.
3.2.7.1 Discussion—
In the context of these test methods, “viability” is understood to be synonymous with cultivability.
4. Summary of Test Methods
NOTE 2—The neutralization test procedure selected must be consistent with the methods of testing used in the antimicrobial effectiveness evaluation.
4.1 Neutralization Assay with Recovery on Semi-solid Medium—Neutralization
...