ASTM E2149-13
(Test Method)Standard Test Method for Determining the Antimicrobial Activity of Immobilized Antimicrobial Agents Under Dynamic Contact Conditions
Standard Test Method for Determining the Antimicrobial Activity of Immobilized Antimicrobial Agents Under Dynamic Contact Conditions
SIGNIFICANCE AND USE
4.1 Chemically bonded, antimicrobial agents are not free to diffuse into their environment under normal conditions of use. This test method ensures good contact between the bacteria and the treated fiber, fabric, or other substrate, by constant agitation of the test specimen in a challenge suspension during the test period.
4.2 The metabolic state of the challenge species can directly affect measurements of the effectiveness of particular antimicrobial agents or concentrations of agents. The susceptibility of the species to particular biocides could be altered depending on its life stage (cycle). One-hour contact time in a buffer solution allows for metabolic stasis in the population. This test method standardizes both the growth conditions of the challenge species and substrate contact times to reduce the variability associated with growth phase of the microorganism.
4.3 Leaching of an antimicrobial is dependent upon the test conditions being utilized and the ultimate end use of the product. For example, water soluble antimicrobials will be prone to removal from the test surface using the method described in Section 13 but insoluble compounds will not. It is for this reason that the use of the term leaching throughout this document is limited to only the testing conditions described herein. To determine if a compound is immobilized in all conditions or during the end use of the product additional testing may be required.
4.4 This test method cannot determine if a compound is leaching into solution or is immobilized on the substrate. This test method is only intended to determine efficacy as described in 4.5 and subsequent portions of the method.
4.5 This test method is intended to evaluate antimicrobial agents that are not removed from the surface by the aqueous testing conditions, as evaluated by Section 13. If an antimicrobial agent that is shown to be removed from the surface by Section 13 is utilized in this test methodology, controls must be inclu...
SCOPE
1.1 This test method is designed to evaluate the antimicrobial activity of non-leaching, antimicrobial-treated specimens under dynamic contact conditions. This dynamic shake flask test was developed for routine quality control and screening tests in order to overcome difficulties in using classical antimicrobial test methods to evaluate substrate-bound antimicrobials. These difficulties include ensuring contact of inoculum to treated surface (as in AATCC 100), flexibility of retrieval at different contact times, use of inappropriately applied static conditions (as in AATCC 147), sensitivity, and reproducibility.
1.2 This test method allows for the ability to evaluate many different types of treated substrates and a wide range of microorganisms. Treated substrates used in this test method can be subjected to a wide variety of physical/chemical stresses or manipulations and allows for the versatility of testing the effect of contamination due to such things as hard water, proteins, blood, serum, various chemicals, and other contaminants.
1.3 Surface antimicrobial activity is determined by comparing results from the test sample to controls run simultaneously.
1.4 The presence of an antimicrobial that requires neutralization is determined by the post-test.
1.5 Proper neutralization of all antimicrobials must be confirmed using Test Methods E1054.
1.6 This test method should be performed only by those trained in microbiological techniques.
1.7 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.
1.8 This standard may involve hazardous materials, operations, and equipment. 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...
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Designation: E2149 − 13
StandardTest Method for
Determining the Antimicrobial Activity of Immobilized
1
Antimicrobial Agents Under Dynamic Contact Conditions
This standard is issued under the fixed designation E2149; 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 address all of the safety concerns, if any, associated with its
use. It is the responsibility of the user of this standard to
1.1 This test method is designed to evaluate the antimicro-
establish appropriate safety and health practices and deter-
bial activity of non-leaching, antimicrobial-treated specimens
mine the applicability of regulatory limitations prior to use.
under dynamic contact conditions. This dynamic shake flask
test was developed for routine quality control and screening
2. Referenced Documents
tests in order to overcome difficulties in using classical
antimicrobial test methods to evaluate substrate-bound antimi- 2
2.1 ASTM Standards:
crobials. These difficulties include ensuring contact of inocu-
E1054Test Methods for Evaluation of Inactivators of Anti-
lum to treated surface (as in AATCC 100), flexibility of
microbial Agents
retrieval at different contact times, use of inappropriately
3
2.2 AATCC Documents:
applied static conditions (as in AATCC 147), sensitivity, and
AATCC 147Antibacterial Activity Assessment of Textile
reproducibility.
Materials: Parallel Streak Method
1.2 This test method allows for the ability to evaluate many
AATCC 100Antibacterial Finishes on Fabrics
different types of treated substrates and a wide range of
microorganisms. Treated substrates used in this test method
3. Summary of Test Method
canbesubjectedtoawidevarietyofphysical/chemicalstresses
3.1 The antimicrobial activity of a substrate-bound, non-
or manipulations and allows for the versatility of testing the
leaching antimicrobial agent is dependent upon direct contact
effect of contamination due to such things as hard water,
of microbes with the active chemical agent. This test deter-
proteins, blood, serum, various chemicals, and other contami-
mines the antimicrobial activity of a treated specimen by
nants.
shaking samples of surface-bound materials in a concentrated
1.3 Surface antimicrobial activity is determined by compar-
bacterial suspension for a one hour contact time. The suspen-
ingresultsfromthetestsampletocontrolsrunsimultaneously.
sion is serially diluted both before and after contact and
1.4 The presence of an antimicrobial that requires neutral-
cultured.The number of viable organisms from the suspension
ization is determined by the post-test.
is determined and the percent reduction (or log reduction) is
10
calculated by comparing retrievals from appropriate controls.
1.5 Proper neutralization of all antimicrobials must be
confirmed using Test Methods E1054.
4. Significance and Use
1.6 This test method should be performed only by those
4.1 Chemically bonded, antimicrobial agents are not free to
trained in microbiological techniques.
diffuse into their environment under normal conditions of use.
1.7 The values stated in SI units are to be regarded as
Thistestmethodensuresgoodcontactbetweenthebacteriaand
standard. No other units of measurement are included in this
thetreatedfiber,fabric,orothersubstrate,byconstantagitation
standard.
of the test specimen in a challenge suspension during the test
1.8 This standard may involve hazardous materials,
period.
operations, and equipment. This standard does not purport to
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
1
This test method is under the jurisdiction of ASTM Committee E35 on contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
Pesticides, Antimicrobials, and Alternative Control Agents and is the direct Standards volume information, refer to the standard’s Document Summary page on
responsibility of Subcommittee E35.15 on Antimicrobial Agents. the ASTM website.
3
Current edition approved July 1, 2013. Published October 2013. Originally Available from American Association of Textile Chemists and Colorists
approved in 2001. Last previous edition approved in 2010 as E2149–10. DOI: (AATCC), P.O. Box 12215, Research Triangle Park, NC 27709, http://
10.1520/E2149-13. www.aatcc.org.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1
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E2149 − 13
4.2 Themetabolicstateofthechallengespeciescandirectly 6.4 Incubator,capableofmaintainingatemperatureof35 6
affect measurements of the effectiveness of particular antimi-
...
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: E2149 − 10 E2149 − 13
Standard Test Method for
Determining the Antimicrobial Activity of Immobilized
1
Antimicrobial Agents Under Dynamic Contact Conditions
This standard is issued under the fixed designation E2149; 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
1.1 This test method is designed to evaluate the antimicrobial activity of non-leaching, antimicrobial-treated specimens under
dynamic contact conditions. This dynamic shake flask test was developed for routine quality control and screening tests in order
to overcome difficulties in using classical antimicrobial test methods to evaluate substrate-bound antimicrobials. These difficulties
include ensuring contact of inoculum to treated surface (as in AATCC 100-2004),100), flexibility of retrieval at different contact
times, use of inappropriately applied static conditions (as in AATCC 147-2004),147), sensitivity, and reproducibility.
1.2 This test method allows for the ability to evaluate many different types of treated substrates and a wide range of
microorganisms. Treated substrates used in this test method can be subjected to a wide variety of physical/chemical stresses or
manipulations and allows for the versatility of testing the effect of contamination due to such things as hard water, proteins, blood,
serum, various chemicals, and other contaminants.
1.3 Surface antimicrobial activity is determined by comparing results from the test sample to controls run simultaneously.
1.4 The presence of a leaching antimicrobial is determined both pre- and an antimicrobial that requires neutralization is
determined by the post-test.
1.5 Proper neutralization of all antimicrobials must be confirmed using Test Methods E1054.
1.6 This test method should be performed only by those trained in microbiological techniques.
1.7 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.
1.8 This standard may involve hazardous materials, operations, and equipment. 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:
E1054 Test Methods for Evaluation of Inactivators of Antimicrobial Agents
3
2.2 AATCC Documents:
AATCC 147-2004147 Antibacterial Activity Assessment of Textile Materials: Parallel Streak Method
AATCC 100-2004100 Antibacterial Finishes on Fabrics
3. Summary of Test Method
3.1 The antimicrobial activity of a substrate-bound, non-leaching antimicrobial agent is dependent upon direct contact of
microbes with the active chemical agent. This test determines the antimicrobial activity of a treated specimen by shaking samples
of surface-bound materials in a concentrated bacterial suspension for a one hour contact time. The suspension is serially diluted
both before and after contact and cultured. The number of viable organisms from the suspension is determined and the percent
reduction (or log reduction) is calculated by comparing retrievals from appropriate controls.
10
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 May 1, 2010July 1, 2013. Published May 2010December 2013. Originally approved in 2001. Last previous edition approved in 20012010 as
E2149 – 01E2149 – 10., which was withdrawn in 2010 and reinstated in May 2010. DOI: 10.1520/E2149-10. DOI: 10.1520/E2149-13.
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or contact ASTM 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.
3
Available from American Association of Textile Chemists and Colorists (AATCC), P.O. Box 12215, Research Triangle Park, NC 27709, http://www.aatcc.org.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1
---------------------- Page: 1 ----------------------
E2149 − 13
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