ASTM E3218-21

Standard Test Method for Quantitative Method for Testing Antimicrobial Agents against Spores of <emph type="ital">C. difficile</emph> on Hard, Nonporous Surfaces

SIGNIFICANCE AND USE
5.1 The test method was designed to determine the LR in spores on a hard, non-porous surface after exposure to a test chemical in a closed system.  
5.2 Each test includes three control carriers (exposed to phosphate buffered saline with Tween-80), three test system control carriers (exposed to 1500 ppm ± 150 ppm sodium hypochlorite), and ten treated carriers (per test chemical/concentration/contact time combination).
SCOPE
1.1 This test method covers a standardized approach to quantitatively determine the effectiveness of antimicrobial chemicals in treating hard, non-porous surfaces contaminated with spores of C. difficile (ATCC 43598) grown in accordance with Practice E2839.  
1.2 This test method is based on principles established for Test Method E2197 and an Organisation for Economic Co-operation and Development Guidance Document.2  
1.3 Training in basic microbiology and aseptic technique are required to perform this assay.  
1.4 Units—The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.5 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.  
1.6 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: Published
• Publication Date: 31-May-2021
• ICS: 07.100.99 - Other standards related to microbiology
• Technical Committee: E35 - Pesticides, Antimicrobials, and Alternative Control Agents
• Drafting Committee: E35.15 - Antimicrobial Agents

Standards Content (Sample)

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.
Designation: E3218 − 21
Standard Test Method for
Quantitative Method for Testing Antimicrobial Agents
1
against Spores of C. difficile on Hard, Nonporous Surfaces
This standard is issued under the fixed designation E3218; 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. Scope* E2197Quantitative Disk Carrier Test Method for Determin-
ing Bactericidal, Virucidal, Fungicidal, Mycobactericidal,
1.1 This test method covers a standardized approach to
and Sporicidal Activities of Chemicals
quantitatively determine the effectiveness of antimicrobial
E2756Terminology Relating toAntimicrobial andAntiviral
chemicals in treating hard, non-porous surfaces contaminated
Agents
with spores of C. diffıcile (ATCC 43598) grown in accordance
E2839Practice for Production and Storage of Spores of C.
with Practice E2839.
difficile for Use in Efficacy Evaluation of Antimicrobial
1.2 This test method is based on principles established for
Agents
Test Method E2197 and an Organisation for Economic Co-
2
2 2.2 Other Standards
operation and Development Guidance Document.
OECD Guidance DocumentQuantitative Method for Evalu-
1.3 Traininginbasicmicrobiologyandaseptictechniqueare
ating Bactericidal Activity of Microbicides used on Hard
required to perform this assay.
Non-Porous Surfaces. Dated June 21, 2013.
1.4 Units—The values stated in SI units are to be regarded
asstandard.Nootherunitsofmeasurementareincludedinthis 3. Terminology
standard.
3.1 Definitions—For definition of terms used in this test
1.5 This standard does not purport to address all of the
method refer to Terminology E2756.
safety concerns, if any, associated with its use. It is the
3.2 Definitions of Terms Specific to This Standard:
responsibility of the user of this standard to establish appro-
3.2.1 frozen spore suspension, n—Apreparation of bacterial
priate safety, health, and environmental practices and deter-
endospores that is maintained at -80°C 65°C.
mine the applicability of regulatory limitations prior to use.
3.2.1.1 Discussion—For the purposes of this test method,
1.6 This international standard was developed in accor-
the definition applies to spore suspensions of C. diffıcile that
dance with internationally recognized principles on standard-
have been prepared and qualified in accordance with Practice
ization established in the Decision on Principles for the
E2839.
Development of International Standards, Guides and Recom-
3.2.1.2 Discussion—Spore suspensions of C. diffıcile used
mendations issued by the World Trade Organization Technical
in this test method may be stored for up to 90 days under the
Barriers to Trade (TBT) Committee.
conditions provided in the definition.
2. Referenced Documents
3.2.2 final test suspension, n—thawed frozen spore suspen-
3
sion (3.2.1) including the addition of a soil load.
2.1 ASTM Standards:
A967Specification for Chemical Passivation Treatments for
3.2.3 test system control, n—a solution of 1500ppm 6150
Stainless Steel Parts
ppm laboratory-grade sodium hypochlorite (NaOCl) used to
validate each efficacy test.
1
This test method is under the jurisdiction of ASTM Committee E35 on
3.3 Acronyms:
Pesticides, Antimicrobials, and Alternative Control Agents and is the direct
responsibility of Subcommittee E35.15 on Antimicrobial Agents.
AISI = American Iron and Steel Institute
CurrenteditionapprovedJune1,2021.PublishedJuly2021.Originallyapproved
CFU = Colony-forming unit
in 2019. Last previous edition approved in 2019 as E3218–19. DOI: 10.1520/
E3218–21
2
Available from the Organisation for Economic Co-operation (OECD) 2, rue
4. Summary of Test Method
André Pascal 75775 Paris Cedex 16, France, www.oecd.org
3
4.1 The method uses disks (1 cm in diameter) of brushed
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
contact ASTM Customer Service at [e-mail]. For Annual Book of ASTM
stainless steel to represent hard, non-porous surfaces.
Standards volume information, refer to the standard’s Document Summary page on
the ASTM website. 4.2 Each disk receives 10 µL of the final test suspension.
*A Summary of Changes section appears at the end of this standard
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1

---------------------- Page: 1 ----------------------
E3218 − 21
4.3 The final test suspension is dried and exposed to 50 µL 7.7 Polyethersulfone membrane filter (PES),forrecoveryof
ofth
...

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.
Designation: E3218 − 19 E3218 − 21
Standard Test Method for
Quantitative Method for Testing Antimicrobial Agents
1
against Spores of C. difficile on Hard, Nonporous Surfaces
This standard is issued under the fixed designation E3218; 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 Scope*
1.1 This test method covers a standardized approach to quantitatively determine the effectiveness of antimicrobial chemicals in
treating hard, non-porous surfaces contaminated with spores of C. diffıcile (ATCC 43598) grown in accordance with Practice
E2839.
1.2 This test method is based on principles established for Test Method E2197 and an Organisation for Economic Co-operation
2
and Development Guidance Document.
1.3 Training in basic microbiology and aseptic technique are required to perform this assay.
1.4 Units—The values stated in SI units are to be regarded as standard. No other units of measurement are included in this
standard.
1.5 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.
1.6 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
3
2.1 ASTM Standards:
A967 Specification for Chemical Passivation Treatments for Stainless Steel Parts
E2197 Quantitative Disk Carrier Test Method for Determining Bactericidal, Virucidal, Fungicidal, Mycobactericidal, and
Sporicidal Activities of Chemicals
E2756 Terminology Relating to Antimicrobial and Antiviral Agents
E2839 Practice for Production and Storage of Spores of C. difficile for Use in Efficacy Evaluation of Antimicrobial Agents
2
2.2 Other Standards
OECD Guidance Document Quantitative Method for Evaluating Bactericidal Activity of Microbicides used on Hard Non-Porous
Surfaces. Dated June 21, 2013.
1
This test method is under the jurisdiction of ASTM Committee E35 on Pesticides, Antimicrobials, and Alternative Control Agents and is the direct responsibility of
Subcommittee E35.15 on Antimicrobial Agents.
Current edition approved Aug. 15, 2019June 1, 2021. Published September 2019July 2021. Originally approved in 2019. Last previous edition approved in 2019 as
E3218–19. DOI: 10.1520/E3218–1910.1520/E3218–21
2
Available from the Organisation for Economic Co-operation (OECD) 2, rue André Pascal 75775 Paris Cedex 16, France, www.oecd.org
3
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.
*A Summary of Changes section appears at the end of this standard
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1

---------------------- Page: 1 ----------------------
E3218 − 21
3. Terminology
3.1 Definitions—For definition of terms used in this test method refer to Terminology E2756.
3.2 Definitions of Terms Specific to This Standard:
3.2.1 frozen spore suspension, n—A preparation of bacterial endospores that is maintained at -80 6 5 °C.-80 °C 6 5 °C.
3.2.1.1 Discussion—
For the purposes of this test method, the definition applies to spore suspensions of C. diffıcile that have been prepared and qualified
in accordance with Practice E2839.
3.2.1.2 Discussion—
Spore suspensions of C. diffıcile used in this test method may be stored for up to 90 days under the conditions provided in the
definition.
3.2.2 final test suspension, n—thawed frozen spore suspension (3.2.1) including the addition of a soil load.
3.2.3 test system control, n—a solution of 1500 6 150 1500 ppm 6 150 ppm laboratory-grade sodium hypochlorite (NaOCl) used
to validate each efficacy test.
3.3 Acronyms:
AISI = American Iron and Steel Institute
CFU = Colony-forming unit
4. Summary of Test Method
4.1 The method uses disks (1 cm in diameter) of brushed st
...